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- Age and growth of megrim, Lepidorhombus boscii, Risso of the portuguese continental coastPublication . Castilho, Rita; Dinis, Maria Teresa; Erzini, KarimThe first study of age and growth of four-spotted scaldfish, more commonly known as megrim (Lepidorhombus boscii, Risso), an important (by catch ) species of the Portuguese trawl fishery from the Atlantic coast is presented. The analysis was based on data obtained from a total of 609 specimens sampled between 1985 and 1986. Otoliths were used for age determination. The Von Bertalanf~, growth parameters were estimated using the non-linear fitting method: Lt (females)=440[ 1 -e -°-14(t+ 1.32) ] and Lt= 375 [ 1 -e -°~4(t+ 1.93) ] for males. L~ and to values are significantly different. The megrim grows allometrically (slope of length-weight regressions > 3 ). Females live longer and gain weight at a faster rate than males. Differences in growth between sexes become more apparent from the second year onwards.
- Replaying the tape: recurring biogeographical patterns in Cape Verde Conus after 12 million yearsPublication . Cunha, R. L.; Tenorio, Manuel J.; Afonso, Carlos; Castilho, Rita; Zardoya, R.Isolated oceanic islands are excellent natural laboratories to test the relative role of historical contingency and determinism in evolutionary diversification. Endemics of the marine venomous snail Conus in the Cape Verde archipelago were originated from at least two independent colonizations of ‘small’ and ‘large’ shelled species separated by 12 million years. In this study, we have reconstructed phylogenetic relationships within large-shelled Conus (C. ateralbus, C. pseudonivifer, C. trochulus, and C. venulatus) based on mitochondrial cox1 and nad4 haplotype sequences. The reconstructed molecular phylogeny revealed three well-supported and relatively divergent clades (A, B, and C) that do not correspond to current species classification based on shell colour and banding patterns. Clade A grouped specimens assigned either to C. pseudonivifer or C. trochulus, clade B is composed of specimens assigned to C. venulatus, and clade C comprises specimens assigned either to C. venulatus or C. ateralbus. Geometric morphometric analyses found significant differences between the radular teeth shape of C. pseudonivifer/C. trochulus and C. venulatus/C. ateralbus. In clades A and B, northwestern Boavista and Maio specimens cluster together to the exclusion of eastern Boavista samples. In Sal, populations form a monophyletic island assemblage (clade C). The large-shelled Conus have remarkably replicated biogeographical patterns of diversification of small-shelled Conus. Similar selective forces (i.e. nonplanktonic lecithotrophy with limited larval dispersal and allopatric diversification) together with repeated instances of low sea level stands during glacial maxima that allowed connection between islands, have overcome the effect of historical contingency, and explain the observed recurring biogeographical patterns.
- Impact of mountain chains, sea straits and peripheral populations on genetic and taxonomic structure of a freshwater turtle, Mauremys leprosa (Reptilia, Testudines, Geoemydidae)Publication . Fritz, U.; Barata, M.; Busack, S. D.; Fritzsch, G.; Castilho, RitaMauremys leprosa, distributed in Iberia and North-west Africa, contains two major clades of mtDNA haplotypes. Clade A occurs in Portugal, Spain and Morocco north of the Atlas Mountains. Clade B occurs south of the Atlas Mountains in Morocco and north of the Atlas Mountains in eastern Algeria and Tunisia. However, we recorded a single individual containing a clade B haplotype in Morocco from north of the Atlas Mountains. This could indicate gene flow between both clades. The phylogenetically most distinct clade A haplotypes are confined to Morocco, suggesting both clades originated in North Africa. Extensive diversity within clade A in south-western Iberia argues for a glacial refuge located there. Other regions of the Iberian Peninsula, displaying distinctly lower haplotype diversities, were recolonized from within south-western Iberia. Most populations in Portugal, Spain and northern Morocco contain the most common clade A haplotype, indicating dispersal from the south-western Iberian refuge, gene flow across the Strait of Gibraltar, and reinvasion of Morocco by terrapins originating in south-western Iberia. This hypothesis is consistent with demographic analyses, suggesting rapid clade A population increase while clade B is represented by stationary, fragmented populations. We recommend the eight, morphologically weakly diagnosable, subspecies of M. leprosa be reduced to two, reflecting major mtDNA clades: Mauremys l. leprosa (Iberian Peninsula and northern Morocco) and M. l. saharica (southern Morocco, eastern Algeria and Tunisia). Peripheral populations could play an important role in evolution of M. leprosa because we found endemic haplotypes in populations along the northern and southern range borders. Previous investigations in another western Palearctic freshwater turtle (Emys orbicularis) discovered similar differentiation of peripheral populations, and phylogeographies of Emys orbicularis and Mauremys rivulata underline the barrier status of mountain chains, in contrast to sea straits, suggesting common patterns for western Palearctic freshwater turtles.
- Genetic structure of Brachidontes puniceus populations in Cape Verde archipelago shows signature of expansion during the last glacial maximumPublication . Cunha, R. L.; Lopes, Evandro P,; Reis, D. M.; Castilho, RitaQuaternary climatic fluctuations had major impacts on species’ distributions over the last 2.5 Myr. Expansions and contractions of the polar ice caps throughout glacial cycles strongly affected terrestrial fauna and flora whereas eustatic sea level variations had major consequences on rocky-shore communities. The effect of these glacial episodes on marine organisms inhabiting oceanic islands is still poorly understood. We analysed the genetic structure of the marine mussel Brachidontes puniceus from the Cape Verde Islands using mitochondrial sequence data. The apparent absence of physical oceanographic barriers or ecological filters in the geographical range of B. puniceus is reflected in the lack of genetic structure found among populations from the entire archipelago. Larval dispersal in B. puniceus likely played a critical role in the demographic connectivity of populations across the Cape Verde archipelago. Results from demographic analysis were consistent with a population expansion promoted by an increase in the habitat available for larval settlement resulting from a low sea-level stand during the last glacial maximum.
- Genetic hypervariability of a Northeastern Atlantic venomous rockfishPublication . Francisco, Sara M.; Castilho, Rita; Lima, Cristina S.; Almada, Frederico; Rodrigues, Francisca; Šanda, Radek; Vukić, Jasna; Pappalardo, Anna Maria; Ferrito, Venera; Robalo, Joana I.Background: Understanding the interplay between climate and current and historical factors shaping genetic diversity is pivotal to infer changes in marine species range and communities’ composition. A hylogeographical break between the Atlantic and the Mediterranean has been documented for several marine organisms, translating into limited dispersal between the two basins. Methods: In this study, we screened the intraspecific diversity of 150 individuals of the Madeira rockfish (Scorpaena maderensis) across its distributional range (seven sampling locations in the Atlantic and Mediterranean basins) using the mitochondrial control region and the nuclear S7 first intron. Results: The present work is the most comprehensive study done for this species, yielding no genetic structure across sampled locations and no detectable Atlantic-Mediterranean break in connectivity. Our results reveal deep and hyper-diverse bush-like genealogies with large numbers of singletons and very few shared haplotypes. The genetic hyper-diversity found for the Madeira rockfish is relatively uncommon in rocky coastal species, whose dispersal capability is limited by local oceanographic patterns. The effect of climate warming on the distribution of the species is discussed.
- Mitochondrial DNA reveals a mosaic pattern of phylogeographical structure in Atlantic and Mediterranean populations of anchovy (Engraulis encrasicolus)Publication . Magoulas, A.; Castilho, Rita; Caetano, Sandra; Marcato, S.; Patarnello, T.This study extends the geographic coverage of a previous study of mitochondrial DNA restriction fragment length polymorphism in European anchovy. Both studies together include 24 samples representing 17 localities extending from the Black Sea, through the Mediterranean Sea to the eastern Atlantic as far south as Dakar, Senegal. Eighty-eight haplotypes deWne two clades (A and B) separated by 3.2% sequence divergence. Clade A has a star-like genealogy indicative of a recent population expansion. Clade B has a more complex genealogy, consisting of several haplotypes at intermediate frequencies. The distributions of these clades consist of a mosaic with abrupt changes between some areas and gradients between other areas. Clade A predominates the Black and Aegean seas, but is present throughout the Mediterranean. Unexpectedly, new data show that clade A is also at a high frequency in the Atlantic, from Portugal to at least Senegal. Overall, the level of genetic diVerentiation among populations is high (FSTD0.148, p<0.0001), with the greatest diVerences between basins. AMOVA reveals four main geographical groups: Atlantic, central Mediterranean, Aegean Sea, and Black Sea. Mismatch distribution clearly indicates historical bottleneck and population expansion for clade A, while for clade B such evidence is equivocal. This diVerence may reXect a range expansion for both clades, but with higher gene Xow (Nm values) between demes for clade A. Both contemporary and historical processes are important in shaping the complex genetic population structure of European anchovy.
- Genetic implications of phylogeographical patterns in the conservation of the boreal wetland butterfly Colias palaeno (Pieridae)Publication . Kramp, Katja; Cizek, Oldrich; Madeira, Pedro M.; Ramos, Ana; Konvicka, Martin; Castilho, Rita; Schmitt, ThomasThe boreo-montane wetland butterfly species Colias palaeno has a European distribution from the Alps to northern Fennoscandia. Within its European range, the species' populations have shrunk dramatically in recent historical times. Therefore, detailed baseline knowledge of the genetic makeup of the species is pivotal in planning potential conservation strategies. We collected 523 individuals from 21 populations across the entire European range and analyzed nuclear (20 allozyme loci) and mitochondrial (600bp of the cytochrome c oxidase subunit I gene) genetic markers. The markers revealed contrasting levels of genetic diversity and divergence: higher in allozymes and lower in mitochondrial sequences. Five main groups were identified by allozymes: Alps, two Czech groups, Baltic countries, Fennoscandia, and Poland. The haplotype mitochondrial network indicates a recent range expansion. The most parsimonious interpretation for our results is the existence of a continuous Wurm glacial distribution in Central Europe, with secondary disjunction during the Last Glacial Maximum into a south-western and a north-eastern fragment and subsequent moderate differentiation. Both groups present signs of postglacial intermixing in the Czech Republic. However, even a complete extinction in this region would not considerably affect the species' genetic basis, as long as the source populations in the Alps and in northern Europe, comprising the most relevant evolutionary units for conservation, are surviving.
- Life in a drop: sampling environmental DNA for marine fishery management and ecosystem monitoringPublication . Gilbey, John; Carvalho, Gary; Castilho, Rita; Coscia, Ilaria; Coulson, Mark W.; Dahle, Geir; Derycke, Sofie; Francisco, Sara M.; Helyar, Sarah J.; Johansen, Torild; Junge, Claudia; Layton, Kara K. S.; Martinsohn, Jann; Matejusova, Iveta; Robalo, Joana I.; Rodriguez-Ezpeleta, Naiara; Silva, Goncalo; Strammer, Ilona; Vasemagi, Anti; Volckaert, Filip A. M.Science-based management of marine fisheries and effective ecosystem monitoring both require the analysis of large amounts of often complex and difficult to collect information. Legislation also increasingly requires the attainment of good environmental status, which again demands collection of data to enable efficient monitoring and management of biodiversity. Such data is traditionally obtained as a result of research surveys through the capture and/or visual identification of organisms. Recent years have seen significant advances in the utilisation of environmental DNA (eDNA) in the marine environment in order to develop alternative cost-effective ways to gather relevant data. Such approaches attempt to identify and/or quantify the species present at a location through the detection of extra-organismal DNA in the environment. These new eDNA based approaches have the potential to revolutionise data collection in the marine environment using non-invasive sampling methods and providing snapshots of biodiversity beyond the capacity of traditional sampling. Here we present a non-technical summary of different approaches in the field of eDNA, and emphasise the broad application of this approach, with value for the governance and management of marine aquatic ecosystems. The review focuses on identifying those tools which are now readily applicable and those which show promise but are currently in development and require further validations. The aim is to provide an understanding of techniques and concepts that can be used by managers without genetic or genomic expertise when consulting with specialists to perform joint evaluations of the utility of the approaches.
- Drivers of Cape Verde archipelagic endemism in keyhole limpetsPublication . Lopes Da Cunha, Regina; Assis, J.; Madeira, Celine; Seabra, Rui; Lima, Fernando P.; Lopes, Evandro P.; Williams, Suzanne T.; Castilho, RitaOceanic archipelagos are the ideal setting for investigating processes that shape species assemblages. Focusing on keyhole limpets, genera Fissurella and Diodora from Cape Verde Islands, we used an integrative approach combining molecular phylogenetics with ocean transport simulations to infer species distribution patterns and analyse connectivity. Dispersal simulations, using pelagic larval duration and ocean currents as proxies, showed a reduced level of connectivity despite short distances between some of the islands. It is suggested that dispersal and persistence driven by patterns of oceanic circulation favouring self-recruitment played a primary role in explaining contemporary species distributions. Mitochondrial and nuclear data revealed the existence of eight Cape Verde endemic lineages, seven within Fissurella, distributed across the archipelago, and one within Diodora restricted to Boavista. The estimated origins for endemic Fissurella and Diodora were 10.2 and 6.7 MY, respectively. Between 9.5 and 4.5 MY, an intense period of volcanism in Boavista might have affected Diodora, preventing its diversification. Having originated earlier, Fissurella might have had more opportunities to disperse to other islands and speciate before those events. Bayesian analyses showed increased diversification rates in Fissurella possibly promoted by low sea levels during Plio-Pleistocene, which further explain differences in species richness between both genera.
- Phylogeography and demographic history of Atherina presbyter (Pisces: Atherinidae) in the North-eastern Atlantic based on mitochondrial DNAPublication . Francisco, S. M.; Castilho, Rita; Soares, M.; Congiu, L.; Brito, A. B.; Vieira, M. N.; Almada, V. C.A fragment of the mitochondrial control region was used to assess phylogeographic patterns and historical demography of the sand-smelt Atherina presbyter in the North-eastern Atlantic, covering its geographical range. A striking result is the highly marked diVerentiation between the Canary Islands population and western European ones. A genetic structure among European populations of A. presbyter was revealed, with a pattern of isolation-by-distance or a gradient eVect at a scale of hundreds kilometres, an uncommon pattern likely related to the biological and life-history traits of the sand-smelt. The northern European populations present a much lower genetic diversity when compared to southern populations, which is consistent with a recent colonization from southern populations. The results showed signs of Pleistocene signatures, with the population age estimates for the European populations being clearly older than the Last Glacial Maximum (18,000 years bp). Nevertheless, paleotemperature reconstructions show that the sand-smelt could not have inhabited the western European shores during the last glacial phase.