Pedro A. Ribeiro

University of Bergen | UiB · Department of Biology

Background. A lutjanid recently captured in Flores Island represents the first documented record of a snapper for the Azores Islands. Since this specimen was not made available to us besides photographs and a muscle sample, a genetic study approach was necessary in order to accurately describe and discuss this somewhat unexpected occurrence. The main objective of this paper was to explain and discuss the presence of this single specimen in this part of the north-eastern Atlantic. Materials and methods. The specimen was caught by spear fishers on 30 July 2014 off Flores Island. We analysed all available photographs and a sample of muscle tissue that was taken and preserved frozen to be used for DNA barcoding. Results. The specimen was a reproductively mature female with a total length of 108.7 cm and weighing 14.84 kg. The body depth of the specimen from the Azores was 2.9 in SL and the analyses conducted showed no divergence from the Canary Islands specimen reported by García-Mederos and Tuset (2014) as Lutjanus dentatus (Duméril, 1861) with the body depth of 2.5 in SL, which demonstrates that this character is highly variable and not useful for species identification. Conclusions. The snapper specimen from the Island of Flores is a Lutjanus cyanopterus (Cuvier, 1828) and confirmed as first record for this part of the north-eastern Atlantic. Comparisons with a L. dentatus from the Canary Islands point towards synonymy although the available molecular evidence is too scarce and by no means conclusive.

The genetic erosion of populations exposed to human exploitation plays a detrimental role on a species ability to adapt to changing environmental conditions. The Macaronesia (NE Atlantic) endemic limpet Patella aspera (Röding 1798) has been subject to overexploitation throughout its geographic distribution. We analysed 841 limpet specimens from eleven islands across the archipelagos of Azores, Madeira and Canaries. Results from 11 nuclear microsatellite markers showed significant population structure between populations from Azores and populations from Madeira and Canaries, and absence of current or historic gene flow between these. M-ratios showed that both population clusters have experienced demographic changes over time. Heterozygote deficits were common across populations, which can be better accounted for by inbreeding than by null alleles or Wahlund effect. Such levels of inbreeding are likely a consequence of a significant reduction of reproductive units due to decades of intensive exploitation. As a sequential protandrous hermaphrodite, the size-selective harvesting of larger individuals likely fosters unbalanced sex-ratios and a consequent reproductive shortage. A recent compensatory hypothesis suggests that males are compensating the removal of larger females by undergoing sex change earlier and presumably at smaller sizes, as an adaptive response of the species under high size-biased fishing pressure. Despite such response, a dramatic reduction of Ne emerging from a large variation in the reproductive success due to overfishing and artificial genetic drift, can simply explain the inbreeding scenario observed in this Macaronesia endemic key species. This study provides valuable insights for management and conservation of P. aspera throughout Macaronesia.

The uptake of natural living resources for human consumption has triggered serious changes in the balance of ecosystems. In the archipelagos of Macaronesia (NE Atlantic), limpets have been extensively exploited probably since islands were first colonized. This has led to profound consequences in the dynamics of rocky shore communities. The Patella candei complex includes various subspecies of limpets that are ascribed to a particular archipelago and has been the focus of several taxonomic surveys without much agreement. Under a conservational perspective, we apply morphometric and genetic analyses to test subspecies boundaries in P. candei and to evaluate its current population connectivity throughout Macaronesia (Azores, Madeira, and Canaries). A highly significant genetic break between archipelagos following isolation by distance was detected (FST = 0.369, p < .001). Contrastingly, significant genetic differentiation among islands (i.e., Azores) was absent possibly indicating ongoing gene flow via larval exchange between populations. Significant shell-shape differences among archipelagos were also detected using both distance-based and geometric morphometric analyses. Adaptive processes associated with niche differentiation and strong barriers to gene flow among archipelagos may be the mechanisms underlying P. candei diversification in Macaronesia. Under the very probable assumption that populations of P. candei from each archipelago are geographically and/or ecologically isolated populations, the various subspecies within the P. candei complex may be best thought of as true species using the denomination: P. candei in Selvagens, Patella gomesii in Azores, Patella ordinaria in Madeira, and Patella crenata for Canaries. This would be in agreement with stock delimitation and units of conservation of P. candei sensu latu along Macaronesia.

Exploitation of organisms can prompt the reduction in the number and size of target populations consequently affecting reproductive output and replenishment. Here, we investigated the effects of exploitation on the population structure of a protandrous patellid limpet, Patella aspera, an overexploited Macaronesian endemic. Timed dives were used to collect animals across eleven islands of Macaronesia. Individuals were inspected for sex, size, and gonad stage. Using catch effort (time per person) per island coastal perimeter as a surrogate for exploitation intensity, we found that limpet abundance (CPUE) and mean size tended to decrease with exploitation intensity. When considering the sex of animals separately, the size of the largest male, but not females, decreased with exploitation. In contrast, the size of the smallest male remained relatively consistent, whereas the size of the smallest female decreased significantly with exploitation. As exploitation is mostly targeting larger individuals, results suggest that males are compensating the removal of larger females, by undergoing sex change at smaller and presumably earlier sizes. These results have wider implications for the conservation of P. aspera, as a reduction in female size will likely affect the numbers of oocytes produced, hence fecundity. Regulations promoting the protection of the larger-sized animals should be enforced to safeguard the replenishment of the population.

The genus Acanthonyx Latreille, 1828 (Majoidea, Epialtidae) contains 17 or 18 known species, depending on competing taxonomic views, that are widely distributed across the world. Morphologically, most species look superficially alike, and therefore, similar taxonomic concepts have been described under different names. Consequently, there is today a considerable list of synonyms, which further complicates the taxonomy of the genus Acanthonyx. In this study, we conducted a phylogeographical and phylogenetic analysis of populations of the genus Acanthonyx in the NE Atlantic, Mediterranean and Macaronesia using the mitochondrial cytochrome oxidase subunit I (COI) and the nuclear 28S rRNA loci. Our phylogenetic and phylogeographical results revealed that Acanthonyx lunulatus sensu lato is a complex of three distinct lineages: one corresponding to the previously described Acanthonyx brevifrons, another to A. lunulatus sensu stricto and a third to a yet undescribed group. Whereas our results confirms that A. brevifrons deserves the status of a species, as it can be easily distinguishable from A. lunulatus by a few morphological traits, we could not find any such traits suitable for the discrimination between A. lunulatus sensu stricto and the third lineage. Furthermore, the degree of COI divergence between this lineage and A. lunulatus is below average levels for Decapoda species. Yet, no shared haplotypes have been detected between them. The differences found in the nuclear gene (indels), together with the sympatric occurrence of the two forms, prompt for a more detailed analysis of this group. Overall, the results show that significant genetic differentiation between specimens with similar morphology occurs in the Epialtidae, thus reinforcing the importance of combining morphological and genetic tools to fully resolve the taxonomy of these decapods.

In the frame of the COST ACTION ‘EMBOS’ (Development and implementation of a pan-European Marine Biodiversity Observatory System), coverage of intertidal macroalgae was estimated at a range of marine stations along the European coastline (Subarctic, Baltic, Atlantic, Mediterranean). Based on these data, we tested whether patterns in macroalgal diversity and distribution along European intertidal rocky shores could be explained by a set of meteo-oceanographic variables. The variables considered were salinity, sea surface temperature, photosynthetically active radiation, significant wave height and tidal range and were compiled from three different sources: remote sensing, reanalysis technique and in situ measurement. These variables were parameterized to represent average conditions (mean values), variability (standard deviation) and extreme events (minimum and maximum values). The results obtained in this study contribute to reinforce the EMBOS network approach and highlight the necessity of considering meteo-oceanographic variables in long-term assessments. The broad spatial distribution of pilot sites has allowed identification of latitudinal and longitudinal gradients manifested through species composition, diversity and dominance structure of intertidal macroalgae. These patterns follow a latitudinal gradient mainly explained by sea surface temperature, but also by photosynthetically active radiation, salinity and tidal range. Additionally, a longitudinal gradient was also detected and could be linked to wave height.

Examining how variability in population abundance and distribution is allotted among different spatial scales can inform of processes that are likely to generate that variability. Results of studies dealing with scale issues in marine benthic communities suggest that variability is concentrated at small spatial scales (from tens of centimetres to few metres) and that spatial patterns of variation are consistent across ecosystems characterized by contrasting physical and biotic conditions, but this has not been formally tested. Here we quantified the variability in the distribution of intertidal rocky shore communities at a range of spatial scales, from tens of centimetres to thousands of kilometres, both in the NE Atlantic and the Mediterranean, and tested whether the observed patterns differed between the two basins. We focused on canopy-forming macroalgae and associated understorey assemblages in the low intertidal, and on the distribution of Patella limpets at mid intertidal levels. Our results highlight that patterns of spatial variation, at each scale investigated, were consistent between the Atlantic and the Mediterranean, suggesting that similar ecological processes operate in these regions. In contrast with former studies, variability in canopy cover, species richness and limpet abundance was equally distributed among spatial scales, possibly reflecting the fingerprint of multiple processes. Variability in community structure of low intertidal assemblages, instead, peaked at the largest scale, suggesting that oceanographic processes and climatic gradients may be important. We conclude that formal comparisons of variability across scales nested in contrasting systems are needed, before any generalization on patterns and processes can be made.

Coastal ecosystems are highly complex and driven by multiple environmental factors. To date we lack scientific evidence for the relative contribution of natural and anthropogenic drivers for the majority of marine habitats in order to adequately assess the role of different stressors across the European seas. Such relationship can be investigated by analysing the correlation between environmental variables and biotic patterns in multivariate space and taking into account non-linearities. Within the framework of the EMBOS (European Marine Biodiversity Observatory System) programme, hard bottom intertidal communities were sampled in a standardized way across European seas. Links between key natural and anthropogenic drivers and hard bottom communities were analysed using Boosted Regression Trees modelling. The study identified strong interregional variability and showed that patterns of hard bottom macroalgal and invertebrate communities were primarily a function of tidal regime, nutrient loading and water temperature (anomalies). The strength and shape of functional form relationships varied widely however among types of organisms (understorey algae composing mostly filamentous species, canopy-forming algae or sessile invertebrates) and aggregated community variables (cover or richness). Tidal regime significantly modulated the effect of nutrient load on the cover and richness of understorey algae and sessile invertebrates. In contrast, hydroclimate was more important for canopy algae and temperature anomalies and hydroclimate separately or interactively contributed to the observed patterns. The analyses also suggested that climate-induced shifts in weather patterns may result in the loss of algal richness and thereby in the loss of functional diversity in European hard bottom intertidal areas.

Background Recent years have seen a rapid increase in survey and sampling expeditions to the Clarion-Clipperton Zone (CCZ) abyssal plain, a vast area of the central Pacific that is currently being actively explored for deep-sea minerals (ISA, 2016). Critical to the development of evidence-based environmental policy in the CCZ are data on the biogeography and connectivity of species at a CCZ-regional level. New information The London Workshop on the Biogeography and Connectivity of the CCZ was convened to support the integration and synthesis of data from European Union (EU) CCZ projects, supported by the EU Managing Impacts of Deep-Sea Resource Exploitation (MIDAS) and EU Joint Programming Initiative Healthy and Productive Seas and Oceans (JPI Oceans) projects. The London Workshop had three clear goals: (1) To explore, review and synthesise the latest molecular biogeography and connectivity data from across recent CCZ cruises from both contractor and academia-funded projects; (2) To develop complementary and collaborative institutional and program-based academic publication plans to avoid duplication of effort and ensure maximum collaborative impact; (3) To plan a joint synthetic data publication highlighting key results from a range of planned molecular biogeography/connectivity publications. 32 participants attended the workshop at the Natural History Museum in London from 10-12 May 2016. Presentations and discussions are summarised in this report covering (1) overviews of current CCZ environmental projects, (2) policy and industry perspectives, (3) synthesis of DNA taxonomy and biogeography studies, (4) summaries of the latest population genetic studies, (5) summaries of the latest broader morphological context, (6) an overview of publication and proposal plans to maximise collaborative opportunities and finally a series of workshop recommendations.

Within the COST action EMBOS (European Marine Biodiversity Observatory System) the degree and variation of the diversity and densities of soft-bottom communities from the lower intertidal or the shallow subtidal was measured at 28 marine sites along the European coastline (Baltic, Atlantic, Mediterranean) using jointly agreed and harmonized protocols, tools and indicators. The hypothesis tested was that the diversity for all taxonomic groups would decrease with increasing latitude. The EMBOS system delivered accurate and comparable data on the diversity and densities of the soft sediment macrozoobenthic community over a large-scale gradient along the European coastline. In contrast to general biogeographic theory, species diversity showed no linear relationship with latitude, yet a bell-shaped relation was found. The diversity and densities of benthos were mostly positively correlated with environmental factors such as temperature, salinity, mud and organic matter content in sediment, or wave height, and related with location characteristics such as system type (lagoons, estuaries, open coast) or stratum (intertidal, subtidal). For some relationships, a maximum (e.g. temperature from 15–20°C; mud content of sediment around 40%) or bimodal curve (e.g. salinity) was found. In lagoons the densities were twice higher than in other locations, and at open coasts the diversity was much lower than in other locations. We conclude that latitudinal trends and regional differences in diversity and densities are strongly influenced by, i.e. merely the result of, particular sets and ranges of environmental factors and location characteristics specific to certain areas, such as the Baltic, with typical salinity clines (favouring insects) and the Mediterranean, with higher temperatures (favouring crustaceans). Therefore, eventual trends with latitude are primarily indirect and so can be overcome by local variation of environmental factors.

Pacific) will impact one of the most remote and least known environments on Earth. Since vast areas are being targeted by concession holders for future mining, large-scale effects of these activities are expected. Hence, insight into the fauna associated with nodules is crucial to support effective environmental management. In this study video surveys were used to compare the epifauna from sites with contrasting nodule coverage in four license areas. Results showed that epifaunal densities are more than two times higher at dense nodule coverage (>25 versus ≤10 individuals per 100 m 2), and that taxa such as alcyonacean and antipatharian corals are virtually absent from nodule-free areas. Furthermore, surveys conducted along tracks from trawling or experimental mining simulations up to 37 years old, suggest that the removal of epifauna is almost complete and that its full recovery is slow. By highlighting the importance of nodules for the epifaunal biodiversity of this abyssal area, we urge for cautious consideration of the criteria for determining future preservation zones.

With anthropogenic impacts rapidly advancing into deeper waters, there is growing interest in establishing deep-sea marine protected areas (MPAs) or reserves. Reserve design depends on estimates of connectivity and scales of dispersal for the taxa of interest. Deep-sea taxa are hypothesized to disperse greater distances than shallow-water taxa, which implies that reserves would need to be larger in size and networks could be more widely spaced; however, this paradigm has not been tested. We compiled population genetic studies of deep-sea fauna, and estimated dispersal distances for 51 studies using a method based on isolation-by-distance slopes. Estimates of dispersal distance ranged from 0.24 km to 2028 km with a geometric mean of 33.2km, and differed in relation to taxonomic and life history factors as well as several study parameters. Dispersal distances were generally greater for fishes than invertebrates with the Mollusca being the least dispersive sampled phylum. Species that are pelagic as adults were more dispersive than those with sessile or sedentary lifestyles. Benthic species from soft substrate habitats were generally less dispersive than species from hard substrate, demersal or pelagic habitats. As expected, species with pelagic and/or feeding (planktotrophic) larvae were more dispersive than other larval types. Many of these comparisons were confounded by taxonomic or other life history differences (e.g. fishes being more dispersive than invertebrates) making any simple interpretation difficult.

The intertidal limpet Patella candei represents an important exploited resource in the Macaronesian archipelagos of Azores, Madeira and Canaries (NE Atlantic). Despite being considered endemic, the taxonomic status of P. candei throughout the region remains unclear. The aim of this study was to develop novel microsatellite markers using next generation sequencing for the limpet P. candei. Twelve novel loci were described and amplified in 103 individuals from two populations (Azores and Madeira) using three multiplex reactions. The number of alleles per locus ranged from 2 to 14 and global observed heterozygosity was 0.417. Genetic differentiation between samples was highly significant (FST?=?0.427) as two main gene pools (K?=?2) were identified using Bayesian approaches. The present genetic tool can be useful to establish the genetic structuring and patterns of connectivity of P. candei in Macaronesia as well as to determine the number of extant subspecies within the P. candei species complex. Such data will provide a fundamental asset to define stocks and thus inform specific conservation strategies that foster the sustainable exploitation of the P. candei complex throughout Macaronesia archipelagos.

The use of three limpet species, Patella vulgata Linnaeus, 1758, Patella depressa Pennant, 1777 and Patella ulyssiponensis Gmelin, 1791 as model organisms in marine ecotoxicology has been evaluated. Initial laboratory experiments were aimed to standardize a biological test with embryos and larvae of Patella spp, establishing the percentage of normal trochophore larvae as endpoint. Before conducting in vitro fertilization, oocytes must be maturated artificially by incubation in an alkaline solution; therefore, alkalinizing agent, pH and time of eggs alkalinization were evaluated. Moreover, time of sperm activation, optimum sperm and oocytes concentration during fertilization, gamete contact time, use of stirring during the fertilization, egg concentration and incubation temperature were examined. Minimum sample size per treatment was also estimated. Exposure of oocytes for 10min to FSW alkalinized with NH4OH at pH 9.0, the use of undiluted sperm pre-activated during 45min and a concentration of 200 oocytesmL(-1), a gamete-contact time of 180min and egg incubation at 18°C during 24h at a concentration of 80 eggsmL(-1) were the conditions allowing maximal embryo-larval development success. With an error of 0.05, a sampling size ≥320 allows a 95% confidence in the estimate. This Patella spp. acute bioassay fulfills a number of important a priori requirements to be used in ecotoxicological studies. Nevertheless, in vitro fertilization requires considerable handling, which may lead to failure in fecundation. Such difficulties are also addressed, in order to facilitate the routine use of this protocol by other laboratories.

Patellid limpets are ecologically important keystone grazers having a long history of overexploitation in the Macaronesian Archipelagos (NE Atlantic islands), where some species, such as Patella aspera, are under serious risk.[1, 2] Patella aspera is a protandric sequential hermaphrodite species with external fertilization, in which individuals start off as males but may undergo a sex reversal with age.[3] Hence, exploitation tends to focus on the larger females in the population as larger limpets (predominantly females) are selectively removed. Despite conservation legislation in Canaries, Madeira and Azores, limpets are under severe pressure and few individuals survive long enough to become females, a phenomenon that severely restricts the effective population size.[4] New conservation actions for the protection and sustainable use of limpets in Macaronesian Archipelagos are urgently needed and should be based on a multidisciplinary framework based on knowledge of the population dynamics and connectivity of this species.

Heteropathes opreski, a new antipatharian species from the northern border of the Oceanographer Fracture Zone is here described and illustrated. An emended diagnosis of the genus and a dichotomous key containing the four Heteropathes species are presented. This species is unique in that it forms smaller colonies compared to the other species in the genus, with some of the lateral pinnules presenting a small ramified subpinnule. Additionally, the polypar spines found on the lateral pinnules are the highest so far recorded in the genus. This record greatly expands the known distribution of this genus, as it was not pre- viously reported to occur in the Northeastern Atlantic.

There is growing consensus that anthropogenic activities significantly disrupt the structure and functioning of marine ecosystems. Oceanic islands such as the Azores (NE Atlantic) are unique habitats with fragile communities, which are highly susceptible to degradation and ecosystem disruption. Patellid limpets have traditionally been collected as a food resource and in 1988 the limpet fishery in São Miguel Island collapsed, after which a one-year ban was implemented allowing the stocks to recover and avoiding catastrophic overexploitation effects. In 1993, legislation was passed to protect this resource, i.e. limpet no-take areas were created, seasonal harvesting restrictions were applied and minimum legal catch sizes were established. However, a recent survey has shown that limpet populations still show clear signs of overexploitation and some populations are virtually extinct in some islands. Here we have developed new multiplexed and described microsatellite markers for the species Patella aspera and P. candei and have examined their genetic diversity, gene flow and population connectivity in the Azores Archipelago. Overall, such information is a fundamental asset to inform conservation strategies and to promote the sustainable exploitation of Macaronesian limpets.

Limpet harvesting in Azores (NE Atlantic) has been taking place probably since the islands were first colonized in the XV century. Limpet species are highly exploited and populations from most islands have declined steadily bringing about catastrophic effects on coastal communities. Despite their economic importance limpets are also ecologically key species and require prioritizing conservation strategies. Patellid limpets are broadcast spawners which go through a planktonic larval stage in their life cycle. They are benthic as adults and the larva is the only phase during their life-cycle which has the ability to disperse over assumed large spatial distances. However, there is now mounting evidence that gene flow between islands and mainlands can be low, even for species with a relatively long planktonic larval stage. Low levels of larval exchange may thus limit the success of conservation objectives expected upon migration and recruitment. In dispersive isolated oceanic islands such as the Macaronesian Islands, the Azores Archipelago in particular, is not clear whether limpet populations from different islands form a single meta-population or, in contrast, populations on each island are isolated from the rest. Knowledge on this scenario is crucial for the management and conservation of exploited populations of limpets. Here we have developed and described species-specific multiplexed microsatellite markers for the limpet Patella aspera using whole genome shotgun 454 sequencing. These genetic tools can allow the study of the population genetic structure and evolutionary history of patellid species in the archipelago of Azores. Genetic studies, alongside with biological, ecological and oceanographic information, represent an important contribution for the understanding of population dynamics by allowing testing hypothesis about larval dispersal patterns, recruitment and life history traits, population connectivity, genetic diversity, and population equilibrium.

The first record of Antipathella subpinnata ( Ellis and Solander, 1786) for the Azores archipelago is presented based on bottom longline by-catch analysis and ROV seafloor surveys, extending the species western-most boundary of distribution in the NE Atlantic. The species was determined using classic taxonomy and molecular analysis targeting nuclear DNA. Although maximum spine height on Azorean colonies branchlets is slightly smaller than that reported from Mediterranean colonies (0.12 vs 0.16 mm), the analysis of partial 18S rDNA, complete ITS1, 5.8S, ITS2 and partial 28S rDNA suggests that the Azorean and Mediterranean specimens belong to the same species. Video surveys of an A. subpinnata garden detected near Pico Island are used to provide the first in situ description of the species habitat in the region and the first detailed description of a black coral garden in the NE Atlantic. With A. subpinnata being the only coral found between 150 and 196 m depths, this is the deepest black coral garden recorded in the NE Atlantic and the first one to be monospecific. The species exhibited a maximum density of 2.64 colonies/m**2 and occurred across a surface area estimated at 67,333 m**2, yielding a local population estimate of 50,500 colonies.

The first record of Antipathella subpinnata (Ellis and Solander, 1786) for the Azores archipelago is presented based on bottom longline by-catch analysis and ROV seafloor surveys, extending the species western-most boundary of distribution in the NE Atlantic. The species was determined using classic taxonomy and molecular analysis targeting nuclear DNA. Although maximum spine height on Azorean colonies branchlets is slightly smaller than that reported from Mediterranean colonies (0.12 vs 0.16 mm), the analysis of partial 18 S rDNA, complete ITS1, 5.8 S, ITS2 and partial 28 S rDNA suggests that the Azorean and Mediterranean specimens belong to the same species. Video surveys of an A. subpinnata garden detected near Pico Island are used to provide the first in situ description of the species habitat in the region and the first detailed description of a black coral garden in the NE Atlantic. With A. subpinnata being the only coral found between 150 to 196 m depths, this is the deepest black coral garden recorded in the NE Atlantic and the first one to be monospecific. The species exhibited a maximum density of 2.64 colonies/m2 and occurred across a surface area estimated at 67,333 m2, yielding a local population estimate of 50,500 colonies.

Limpet harvesting in Azores (NE Atlantic) has been taking place probably since the islands were first colonized. These species are highly exploited and the stocks in most islands have declined steadily with catastrophic effects on coastal communities. They are a locally important resource but also ecologically important species and require prioritizing conservation strategies. Here we describe and develop species-specific microsatellite markers for the limpets Patella candei and Patella aspera using whole genome shotgun 454 sequencing. A subset of 22 and 15 polymorphic microsatellite markers are described for P. aspera and P. candei, respectively. These novel genetic markers can be used to study the population genetic structure and evolutionary history of both patellid species and thus to contribute to stock conservation and management along their distributional area.

We have conducted an exhaustive compilation of records of alcyonaceans, antipatharians, scleractinians and stylasterids available through present day to assess the diversity, distribution and spatial structure of coral assemblages in the Azores exclusive economic zone (EEZ). The resulting database comprises 2501 entries concerning historical oceanographic ex-peditions and other published sources, as well as unpublished data from bottom longline bycatch. Our taxonomic inventory appears to be fairly complete for the explored habitats, accounting for 164 species (79 alcyonaceans, 58 scleractinians, 18 antipatharians and 9 stylasterids), nine of which were documented for the first time. The Azores EEZ harbours a mixed coral fauna with several zoogeographic origins, showing the closest affinity with the Lusitanian–Mediterranean region. Very few apparent endemics were found (14 %), and only in part supported by consistent sampling. Coral diversity is particularly high between 300 and 900 m depths, in areas recognized as traditional fishing grounds or exploitable fish habitat within the 100-mile limit of the EEZ. The composition of coral assemblages shows significant geographical structure among longitudinal sections of the study area at comparable depths (100–1500 m). There is no evidence of a possible role of the Mid-Atlantic Ridge or latitudinal effects underlying this pattern, which suggests that it may instead reflect assemblage variability among features. Stronger changes in species composition were found along the bathymetric gradient. Notwithstanding the mix of partially overlapping steno-and eurybathic species that characterize the vertical distribution of corals, there is a distinct transition from shallow (100–600 m) to intermediate (600–1000 m) depths. The analysis presented here constitutes a valuable contribution for efficient conservation policies of coral-associated vulnerable marine ecosystems and their sustainable use as fishing areas.

There is a growing consensus that over-fishing can have profound community-level effects particularly when keystone species are targeted. Patellid limpets, for instance, have traditionally been collected as a food resource throughout the Macaronesian archipelagos. In the Azores, recent work has shown that current legislation has been largely ineffective in protecting their populations. These organisms are highly exploited and stocks have been declining steadily in most islands with catastrophic effects on coastal communities. For conservation purposes it is important to examine the degree to which populations of exploited patellid species are connected in a dispersive system such as the Macaronesia islands. Research should then address the genetic variation, connectivity, and resilience of limpet populations in Macaronesia, particularly in the Azores. The coupling of genetic biological, ecological and oceanographic information, will decisively contribute for the understanding of population dynamics by allowing the test of hypotheses about larval dispersal patterns, recruitment and life history traits, population connectivity, genetic diversity, and population equilibrium. This multidisciplinary approach will provide information of theoretical and practical importance that may be used to inform conservation strategies and promote the sustainable exploitation of limpets in Macaronesia, particularly in the Azores.

Cold-water corals are widely considered as important structural components of benthic habitats, potentially enhancing local abundance in a variety of fish and invertebrate species. Yet, current knowledge of the taxonomic diversity and distribution patterns of these vulnerable, slow-growing organisms is scarce and fragmented, limiting the effectiveness of spatial management and conservation measures. We have conducted an exhaustive compilation of records of alcyonaceans, antipatharians, scleractinians and stylasterids available until the present day to assess the diversity, distribution, and spatial structure of coral assemblages in the Azores Exclusive Economic Zone (EEZ). The resulting database comprises 2501 entries concerning historical oceanographic expeditions and other published sources, as well as recent data from longline bycatch. Our taxonomic inventory appears to be fairly complete for the explored habitats, accounting for 164 species (79 alcyonaceans, 58 scleractinians, 18 antipatharians and 9 stylasterids), nine of which were new records. The Azores EEZ harbours a mixed coral fauna with several zoogeographic origins, showing the closest affinity with the Lusitanian– Mediterranean region. Very few apparent endemics were found (14%), and only in part supported by consistent sampling. Coral diversity is particularly high between 300 and 900 m depths, in areas recognized as traditional fishing grounds or exploitable fish habitat within the 100-mile limit of the EEZ. The composition of coral assemblages shows significant geographical structure among longitudinal sections of the study area at comparable depths (100–1500 m). There is no evidence of a possible role of the Mid- Atlantic Ridge or latitudinal effects underlying this pattern, which suggests that it may instead reflect assemblage variability among features. Stronger changes in species composition were found along the bathymetric gradient. Notwithstanding the mix of partially overlapping steno- and eurybathic species that characterize the vertical distribution of corals, there is a distinct transition from shallow (100–600 m) to intermediate (600–1000 m) depths. The analysis here presented constitutes a valuable contribution for efficient conservation policies of coral-associated vulnerable marine ecosystems and their sustainable use as fishing areas.

Aim The Lusitanian limpet, Patella rustica Linnaeus, 1758, is a rocky shore intertidal mollusc found throughout the Mediterranean and the north-east Atlantic from Mauritania to southern France. A 280-km gap in its distribution in the Iberian north-west was first reported in the early 20th century. Between 2002 and 2005 this gap was bridged, and unusual climatic and hydrographic events that occurred in the late 1990s are hypothesized to have facilitated transport and settlement of colonizers from the southern edge of the gap. In the present work we used genetic data to investigate differentiation between the edges of the former gap, to determine the origin of colonizers and to assess patterns of connectivity across the historical and newly colonized region.

The reproductive cycles of four limpet species, Patella depressa, Patella ulyssiponensis, Patella vulgata and Patella rustica are described for several locations in northern and central Portugal. Patella depressa and P. ulyssiponensis displayed almost con- tinual gonad activity, with two main spawning periods, September–January and March–June, and minor spawning events dispersed throughout the year. Patella vulgata and P. rustica exhibited highly synchronized reproductive cycles, with well- defined breeding and resting periods. Gonad development in P. vulgata started in September and spawning took place from November–December to March. In P. rustica development started in June and spawning occurred from September– October to December–January. Compared to the 1980s these results suggest that breeding periods are becoming longer in southern species and shorter in northern species.

The occurrence of the pod weed, Halidrys siliquosa, is recorded for the first time on the Portuguese coast. Several specimens of this brown algae were observed attached to the rocky surface in tide pools at 41º44′10″N 8º52′34″W, extending southward its previously known geographical distribution. The observed shift is inconsistent with general predictions of species migrations under warming climate conditions, which anticipate poleward shifts rather than southern expansions. Although more data will be required to undoubtedly uncover its cause, the recently observed range expansion raises important questions about the generalization of the previously stated biogeographic rules.

Dispersal and connectivity of patellid limpets (Patella spp.) in the eastern North Atlantic have been examined by addressing reproductive biology, larval development, population genetics and physical modelling of dispersal. The reproductive cycles of four limpet species were assessed on the northern and central Portugese coast, to determine spawning periods. This information was incorporated into dispersal models. The results showed that P. depressa and P. ulyssiponensis have almost year-round breeding, with a brief resting phase in the early summer. Conversely, the two other species displayed much shorter spawning periods, with gamete release taking place between December and March in P. vulgata and between September and December in P. rustica. The relationship between temperature and planktonic periods in P. depressa, P. ulyssiponensis, and P. vulgata was investigated with laboratory rearing experiments. Average duration of precompetent periods varied inversely with temperature, ranging between 3.7-14.0 days in P. depressa, 2.8-13.7 days in P. ulyssiponensis and 5.7-14.6 days in P. vulgata, whilst delay periods ranged between 15.8-25.4 days in P. depressa, 14.5-27 days in P. ulyssiponensis and 16.5-25 days in P. vulgata. Population genetic structure was examined on a range-wide scale in P. depressa and along the Iberian coast in P. rustica using microsatellite markers, plus one mtDNA locus in P. rustica. Results suggested high levels of gene flow throughout the study area and widespread lack of population differentiation in both species. A biophysical model of dispersal has been developed to assess the degree of demographic connectivity over ecological and evolutionary time frames, and to identify possible barriers to dispersal for P. depressa and P. rustica. The model predicted high levels of connectivity through most of the study area in both species, but in P. depressa simulations identified two large extensions of adult habitat discontinuity as barriers to larval dispersal. The model also showed that despite the potential for long-distance dispersal, most of the larvae released at one given location settle within much shorter distances. These results illustrate the need to view the study of marine dispersal as a multidisciplinary task, and suggest that relying on just one line of evidence may produce misleading results.

Eighteen microsatellite markers have been characterized from the Patella rustica genome. An average of 10 alleles per locus and an expected heterozygosity ranging from 0.054 to 0.937 were observed in a sample of 32 wild individuals from Viana do Castelo, Portugal. Distinct cross-priming amplification rates were recovered on four additional Patella species. Three monomorphic P. rustica loci were polymorphic in other Patella species. The microsatellites developed herein could be a useful intraspecific genetic tool to undertake fine population studies in the genus Patella.

The occurrence of longfin mako, Isurus paucus is recorded for the first time in the mid-North Atlantic. Two specimens were caught, including a 215 cm stretched total length mature male, at 42°50′N 36°16′W by a commercial longline vessel targeting swordfish.

Well-documented changes in species abundances and distributions coinciding with global warming have been increasing during recent years. A trend of raising sea-surface temperature has also been observed along the Portuguese coast which could affect intertidal species' ranges. The present study aimed at evaluating the direction and intensity of distribution changes of macroalgae in the area. The last 50-year trend of coastal air and sea temperature was reassessed, providing an accurate estimate of the warming process. Information on species' range shifts was obtained by comparing data from recent resurveys with historical records of algal distributions collected during the 1950s and 1960s. Although a prevalence of northward migrations was anticipated, this work showed a marked difference in the average direction of changes between cold- and warm-water species. Cold-water species, when considered together, showed no particular shifting trend, because the number of species that shifted north or south was the same. Contrarily, all shifting warm-water species expanded their range northwards. Therefore, generalizations about poleward range shifts due to increasing temperature should be made with caution.

A climate envelope approach was used to model the distributions of the intertidal gastropod Patella rustica, to test the robustness of forecast responses to climate change. The model incorporated variables that were likely to determine the abundance and the northern range limit of this species in the NE Atlantic. The model was built using classification and regression tree analysis (CART) trained with historical distribution data from the mid 1950s and a set of corresponding climatic and oceanographic variables. Results indicated air and sea temperature, in particular during the reproductive and settlement periods, as the main determinants of the Atlantic distribution of P. rustica. The model was subsequently fed with contemporary climatic data and its output was compared with the current distribution and abundance of P. rustica, assessed during a 2002–2003 survey. The model correctly hindcasted the recent collapse of a distributional gap in northern Portugal, as well as an increase in abundance at locations within its range. The predicted northward expansion of the northern range limit did not occur because the absence of the species was confirmed in a survey encompassing the whole Atlantic French coast up to Brest. Stretches of unsuitable habitat too long to be overcome by dispersal are the likely mechanism controlling the northern limit of the distribution of this intertidal species.

Limpets (Patella spp.) are suitable organisms to investigate the effects of climate change in marine systems. They are widespread over NE Atlantic intertidal rocky shores and have been extensively studied in terms of population dynamics and ecology. Within genus Patella, microsatellites have only been developed for Patella caerulea and cross-species tests are unknown. In this work, we describe 11 primer pairs for Patella depressa and the results of cross-species testing on Patella candei and Patella rustica.

Aim Recent colonization of northern Portuguese shores by Patella rustica Linnaeus, 1758, led to the bridging of a historical gap in the distribution known since the 1900s. Long-term oceanographic data collected over the last half-century were examined in order to detect possible mechanisms for the observed change in its distribution. Location This study was carried out along the entire Portuguese coastline, from 41°50′ to 37°06′ N. Time-series of hydrographical variables (sea surface temperature and salinity) were derived for the Atlantic coast of the Iberian Peninsula. Methods Abundance and size-frequency distributions of the newly observed limpet populations were compared with those from well-established populations in southern Portugal. Anomalies were computed for sea surface temperature (1950–2000) and sea surface salinity (1958–2001) data, covering the whole Atlantic coast of the Iberian Peninsula. An upwelling index (1967–2005) was derived for a single location within the distributional gap of P. rustica. Split moving window analysis was performed to detect significant discontinuities in hydrographical data sets. Results Patella rustica has gradually been expanding in northern Iberia, and in the late 1990s the historical gap in distribution in northern Portugal was bridged. Size-frequency distribution differed between historical and recent populations, the latter lacking small-sized individuals. At the same time, several anomalous oceanographic events occurred off the Portuguese coast and were probably related to this expansion. Main conclusions Although sea surface temperature might be a major determinant of the reproductive success of P. rustica and hence its dispersal potential, it is more likely that a coincidence of several factors occurring in the late 1990s provided exceptional conditions that allowed the geographical expansion of this species.

Geographical patterns in the distribution of epifaunal crustaceans (Amphipoda, Isopoda and Tanaidacea) occurring with dominant macroalgal species were investigated along the Portuguese rocky coast. Three regions, each encompassing six shores, were studied. Algal species were selected according to their geographical distribution: Mastocarpus stellatus and Chondrus crispus (north); Bifurcaria bifurcata (north-centre); Plocamium cartilagineum and Cystoseira tamariscifolia (centre-south); Corallina spp. and Codium tomentosum (entire coast). Multivariate techniques were used to test for differences in crustacean assemblage composition between sub-regions and host algal species. A clear gradient of species substitution was observed from north to south. Differences in abundance and diversity of epifaunal crustaceans were observed between southern locations and the remaining sites. Four species were recorded for the first time in the Portuguese coast. Among the 57 taxa identified, southern distribution limits were observed for three species and northern distribution limits were observed for four species. Interestingly, the observed geographical patterns in epifaunal abundance and diversity were not related with geographical changes in the indentity of the dominant algal species.

Environmental impact assessment of unplanned disturbances is often difficult to accomplish due to the absence of ‘before’ data for the impacted sites. In an attempt to overcome this problem, a beyond BACI model is used in order to detect possible changes in the temporal patterns of variation when no previous data are available. The model attempted to detect changes in the abundance of macroinvertebrate species inhabiting the intertidal mussel matrix after an oil spill which occurred in northern Portugal. The detection of a significant impact failed, most probably due to low temporal replication. An extension of the analysis, including the hierarchical arrangement of temporal variability in periods, suggests that increasing the number of sampling times may result in a higher efficiency of the model.

Geographical patterns in the distribution of epifaunal crustaceans (Amphipoda, Isopoda and Tanaidacea) occurring with dominant macroalgal species were investigated along the Portuguese rocky coast. Three regions, each encompassing six shores, were studied. Algal species were selected according to their geographical distribution: Mastocarpusstellatus and Chondrus crispus (north); Bifurcariabifurcata (north-centre); Plocamiumcartilagineum and Cystoseiratamariscifolia (centre-south); Corallina spp. and Codiumtomentosum (entire coast). Multivariate techniques were used to test for differences in crustacean assemblage composition between sub-regions and host algal species. A clear gradient of species substitution was observed from north to south. Differences in abundance and diversity of epifaunal crustaceans were observed between southern locations and the remaining sites. Four species were recorded for the first time in the Portuguese coast. Among the 57 taxa identified, southern distribution limits were observed for three species and northern distribution limits were observed for four species. Interestingly, the observed geographical patterns in epifaunal abundance and diversity were not related with geographical changes in the indentity of the dominant algal species.

Environmental impact assessment of unplanned disturbances is often difficult to accomplish due to the absence of ‘before’ data for the impacted sites. In an attempt to overcome this problem, a beyond BACI model is used in order to detect possible changes in the temporal patterns of variation when no previous data are available. The model attempted to detect changes in the abundance of macroinvertebrate species inhabiting the intertidal mussel matrix after an oil spill which occurred in northern Portugal. The detection of a significant impact failed, most probably due to low temporal replication. An extension of the analysis, including the hierarchical arrangement of temporal variability in periods, suggests that increasing the number of sampling times may result in a higher efficiency of the model.

A shark tagging programme along the Portuguese coast was initiated in 2001 in collaboration with the National Marine Fisheries Service. From a total of 168 blue sharks (Prionace glauca) tagged, 34 sharks were recaptured (20% return rate) providing important information on this species' movement patterns for the area. A total of 28 sharks travelled less than 1000 km while at liberty for time periods ranging from 22 to 1294 days. The remaining fish travelled long distances to north-west Africa, central Atlantic and the Bay of Biscay. Only one shark made a transatlantic migration, being recaptured 3187 km from the tagging site. North–south movements seem to be related to seasonal sea-surface temperature variation in the north-east Atlantic. Seasonal segregation of different life stages also occurs.

14 15 Running Head 16 Changes in the distribution of Patella. rustica 17 18 ABSTRACT 19 20 Aim Recent colonization of northern Portuguese shores by Patella rustica Linnaeus, 1758, led to 21 the bridging of an historical distributional gap known since the 1900s. Long-term oceanographic