Molecular Detection of <Emphasis Type="Italic">Xenostrobus securis</Emphasis> and <Emphasis Type="Italic">Mytillus Galloprovincialis</Emphasis> Larvae in Galician Coast (Spain)

The mussel species Xenostrobus securis from New Zealand was detected in the Spanish coast recently, in the mouth of the Verdugo River into the Vigo Ria. In view of the great importance of the farm mussel sector in this region, the presence of this alien species greatly concerned producers and administration authorities, because of its potential medium- or long-term effects on the autochthonous species, Mytilus galloprovincialis, an important marine resource widely exploited in this location. The goal of this study was to develop a DNA-based technique to identify X. securis and M. galloprovincialis larvae in plankton samples, which would allow monitoring for the presence of X. securis in different points of the Vigo Ria. The techniques used were simplex and multiplex polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and fragment analysis. The application of this system to planktonic samples could be an effective means to assess the presence of the alien species, allowing monitoring if its dispersion is increasing, or on the contrary, if its distribution is restricted to the mouth of the Verdugo River, where X. securis was first detected. In addition, the application of this system at different times could be useful to assess the presence of larvae of these two species in the plankton.     

Invasive and non-invasive lineages in Xenostrobus (Bivalvia: Mytilidae)

The mussel genus Xenostrobus comprises eight named extant species from the Indo-West Pacific, including Xenostrobus inconstans, Xenostrobus pulex and Xenostrobus securis from Australia, the latter two also occurring in New Zealand. Xenostrobus species are predominantly restricted to estuaries or sheltered marine habitats although X. pulex inhabits the rocky intertidal on open coasts. Xenostrobus has recently been suggested to be congeneric with the freshwater invasive species Limnoperna fortunei. Xenostrobus securis is itself invasive in the Far East and Europe. This study employed DNA sequences from cytochrome c oxidase (COI), the D1 expansion region of 28S rRNA and the internal transcribed spacers of the ribosomal cistrons to investigate species relationships and habitat transitions in Australasian Xenostrobus. It is unlikely that Xenostrobus and Limnoperna are congeneric as their COI sequences are very different. There was unexpected complexity in defining Xenostrobus species. Xenostrobus pulex from New Zealand is probably a distinct species to the Australian taxon of that name with the name X. neozelanicus available for that taxon. Xenostrobus inconstans and Australian X. pulex were not reciprocally monophyletic in COI analyses. The phylogeography of the COI haplotypes of X. securis suggests that it may be a cryptic species complex, although this possibility could not be confirmed by sequences of the other genes.     

Impacts of bottom and suspended cultures of mussels <Emphasis Type="Italic">Mytilus</Emphasis> spp. on the surrounding sedimentary environment and macrobenthic biodiversity

The aim of this study was to quantify the effect of bottom and suspended mussel cultures, cultured in different physical environments, on the sedimentary environmental conditions and thereby the biodiversity structure of the associated macrofaunal community. We compared two bottom cultures (Limfjorden: microtidal, wind-driven; Oosterschelde: macrotidal) and one suspended culture (Ria de Vigo in an upwelling coastal region). The sedimentary environmental conditions (mud fraction, POC, PON, phosphorus content, chl a breakdown products) were significantly elevated underneath and surrounding bottom and suspended cultures compared to culture-free sediments that were nearby and hydrodynamically similar. The relative change in environmental conditions was more pronounced in the Oosterschelde compared to Limfjorden, most likely due to differences in hydrodynamic forcing and characteristics of the mussel bed. The effect of the suspended cultures in Ria de Vigo on the surrounding sediments was influenced by local topographic and hydrodynamic conditions. The impact of mussels on the benthic community due to biodeposition was clearly seen in the community structure. The species composition changed from species which are typically present in sandy environments to more small opportunistic species, which are typically present in organically enriched sediments. The impact of bottom cultures on the benthic community due to changes in the habitat under the presence of mussels was positive, especially in the Oosterschelde where an increase in the number of epibenthic species was seen. The influence of bottom cultures on the sedimentary environment and on the macrobenthic community seems to be very local. Within the mussel site in Limfjorden, differences were detected between sites where none or almost no mussels were present with sites where mussels were very abundant.     

DNA barcoding for assessment of exotic molluscs associated with maritime ports in northern Iberia

Ports are gateways for aquatic invasions. New arrivals from maritime traffic and disturbed environmental conditions can promote the settlement of exotic species. Molluscs fall into the most prevalent group of invasive species and can have a tremendous impact on aquatic ecosystems. Here we have investigated exotic molluscs in three ports with different intensities of maritime traffic in the Cantabrian Sea. DNA barcodes were employed to identify the species using BLASTn and BOLD IDS assignment. Deep morphological analysis using diagnostic criteria confirmed BLAST species assignation based on COI and 16S rRNA genes. Results confirmed the usefulness of DNA barcoding for detecting exotic species that are visually similar to native species. Three exotic bivalves were identified: Ostrea stentina (dwarf oyster), the highly invasive Crassostrea gigas (Pacific oyster) and Xenostrobus securis (pygmy mussel). This is the first record of O. stentina in the Bay of Biscay and the second of X. securis in the Cantabrian Sea. Furthermore, we report on the presence of the cryptogenic mussel Mytilaster minimus in the central Cantabrian Sea. These exotic species might have been overlooked due to their phenotypic similarity with co-occurring oyster and mussel species. This study illustrates how combining morphological and DNA taxonomic analysis can help in port and marina biosecurity surveys.     

Fine-scale spatial partitioning of genetic variation and evolutionary contestability in the invasive estuarine mussel Xenostrobus securis

Initial studies of Australian populations of the widely invasive mussel Xenostrobus securis raised the possibility of non-random spatial partitioning within estuaries of mitochondrial DNA (mtDNA) clades that have dispersed intercontinentally and those that have not. A fine-scale phylogeographic investigation was made to examine this possibility using cytochrome c oxidase DNA sequences and data from five microsatellite loci developed here. X. securis in the study region was found to comprise multiple robustly supported mtDNA clades, many of which were genetically very distinct. Frequently, the clades comprised numerous haplotypes that have narrow ranges and may have evolved in situ within drainages or nearby. Microsatellite data reveal significant intra- and inter-drainage differentiation but at lower levels than mtDNA. Variation in mtDNA appears to reflect not only recolonization after infrequent local extinction within drainages but also the low probability of migratory haplotypes successfully establishing during evolutionary contests, with resident haplotypes having selective or demographic advantages derived from local occupation. The patterns of mtDNA haplotype distribution suggest that central New South Wales is the source of the internationally invasive lineages of X. securis.     

Byssus attachment strength of two mytilids in mono-specific and mixed-species mussel beds

The mussel Xenostrobus securis is endemic to the brackish waters of New Zealand and Australia, but has successfully invaded the inner Galician Rías of NW Spain, where it coexists with the indigenous mussel Mytilus galloprovincialis. In this laboratory study, the plasticity of the byssus attachment strength of two mytilids was compared by manipulating substratum, salinity, and bed assembly. M. galloprovincialis showed stronger byssus detachment strength than X. securis, despite lower byssus coverage. Both species responded similarly to the substratum, with substantially lower byssus strength on methacrylate, which offered the lowest surface free energy. Byssus detachment values for M. galloprovincialis were lower at lower salinity. In mixed beds, a number of mussels moved upwards, eventually colonising the upper layers of the assemblage. This behaviour increased byssus strength but only for X. securis. X. securis is adapted to a wide spectrum of abiotic conditions, a trait that may promote its dissemination within estuarine environments.     

Accuracy in the estimation of quantitative minimal area from the diversity/area curve

The problem of representativity is fundamental in ecological studies. A qualitative minimal area that gives a good representation of species pool [C.M. Bouderesque, Methodes d'etude qualitative et quantitative du benthos (en particulier du phytobenthos), Tethys 3(1) (1971) 79] can be discerned from a quantitative minimal area which reflects the structural complexity of community [F.X. Niell, Sobre la biologia de Ascophyllum nosodum (L.) Le Jolis en Galicia, Invest. Pesq. 43 (1979) 501]. This suggests that the populational diversity can be considered as the value of the horizontal asymptote corresponding to the curve sample diversity/biomass [F.X. Niell, Les applications de l'index de Shannon a l'etude de la vegetation interdidale, Soc. Phycol. Fr. Bull. 19 (1974) 238]. In this study we develop a expression to determine minimal areas and use it to obtain certain information about the community structure based on diversity/area curve graphs. This expression is based on the functional relationship between the expected value of the diversity and the sample size used to estimate it. In order to establish the quality of the estimation process, we obtained the confidence intervals as a particularization of the functional (h-phi)-entropies proposed in [M. Salicru, M.L. Menendez, D. Morales, L. Pardo, Asymptotic distribution of (h,phi)-entropies, Commun. Stat. (Theory Methods) 22 (7) (1993) 2015]. As an example used to demonstrate the possibilities of this method, and only for illustrative purposes, data about a study on the rocky intertidal seawed populations in the Ria of Vigo (N.W. Spain) are analyzed [F.X. Niell, Estudios sobre la estructura, dinamica y produccion del Fitobentos intermareal (Facies rocosa) de la Ria de Vigo. Ph.D. Mem. University of Barcelona, Barcelona, 1979].     

Molecular ecology of zebra mussel invasions

The invasion of the zebra mussel, Dreissena polymorpha, into North American waters has resulted in profound ecological disturbances and large monetary losses. This study examined the invasion history and patterns of genetic diversity among endemic and invading populations of zebra mussels using DNA sequences from the mitochondrial cytochrome oxidase I (COI) gene. Patterns of haplotype frequency indicate that all invasive populations of zebra mussels from North America and Europe originated from the Ponto-Caspian Sea region. The distribution of haplotypes was consistent with invasive populations arising from the Black Sea drainage, but could not exclude the possibility of an origin from the Caspian Sea drainage. Similar haplotype frequencies among North American populations of D. polymorpha suggest colonization by a single founding population. There was no evidence of invasive populations arising from tectonic lakes in Turkey, while lakes in Greece and Macedonia contained only Dreissena stankovici. Populations in Turkey might be members of a sibling species complex of D. polymorpha. Ponto-Caspian derived populations of D. polymorpha (theta = 0.0011) and Dreissena bugensis (one haplotype) exhibited low levels of genetic diversity at the COI gene, perhaps as a result of repeated population bottlenecks. In contrast, geographically isolated tectonic lake populations exhibited relatively high levels of genetic diversity (theta = 0.0032 to 0.0134). It is possible that the fluctuating environment of the Ponto-Caspian basin facilitated the colonizing habit of invasive populations of D. polymorpha and D. bugensis. Our findings were concordant with the general trend of destructive freshwater invaders in the Great Lakes arising from the Ponto-Caspian Sea basin.     

DNA barcodes for globally threatened marine turtles: a registry approach to documenting biodiversity

DNA barcoding is a global initiative that provides a standardized and efficient tool to catalogue and inventory biodiversity, with significant conservation applications. Despite progress across taxonomic realms, globally threatened marine turtles remain underrepresented in this effort. To obtain DNA barcodes of marine turtles, we sequenced a segment of the cytochrome c oxidase subunit I (COI) gene from all seven species in the Atlantic and Pacific Ocean basins (815 bp; n = 80). To further investigate intraspecific variation, we sequenced green turtles (Chelonia mydas) from nine additional Atlantic/Mediterranean nesting areas (n = 164) and from the Eastern Pacific (n = 5). We established character-based DNA barcodes for each species using unique combinations of character states at 76 nucleotide positions. We found that no haplotypes were shared among species and the mean of interspecific variation ranged from 1.68% to 13.0%, and the mean of intraspecific variability was relatively low (0–0.90%). The Eastern Pacific green turtle sequence was identical to an Australian haplotype, suggesting that this marker is not appropriate for identifying these phenotypically distinguishable populations. Analysis of COI revealed a north–south gradient in green turtles of Western Atlantic/Mediterranean nesting areas, supporting a hypothesis of recent dispersal from near equatorial glacial refugia. DNA barcoding of marine turtles is a powerful tool for species identification and wildlife forensics, which also provides complementary data for conservation genetic research.     

Zebra mussels (Dreissena polymorpha) in Ireland, AFLP-fingerprinting and boat traffic both indicate an origin from Britain

1. The zebra mussel (Dreissena polymorpha) is an aquatic nuisance species that invaded Ireland around 1994. We studied the invasion of the zebra mussel combining field surveys and genetic studies, to determine the origin of invasion and the vector of introduction. 2. Field surveys showed that live zebra mussels, attached to the hulls of pleasure boats, were transported from Britain to Ireland. These boats were lifted from British waters onto trailers, transported to Ireland by ferry and lifted into Irish waters within a day. Length‐frequency distributions of dead and living mussels on one vessel imported 3 months earlier revealed a traumatic occurrence caused by the overland, air‐exposed transportation. Results show that a large number of individuals survived after re‐immersion in Irish waters and continued to grow. 3. Zebra mussels from populations in Ireland, Great Britain, the Netherlands, France and North America, were analysed using amplified fragment length polymorphisms (AFLP)‐fingerprinting to determine the origin of the Irish invasion. Phylogenetic analysis revealed that Irish and British mussels clustered closely together, suggesting an introduction from Britain. 4. Ireland remained un‐invaded by the zebra mussel for more than 150 year. The introduction of the zebra mussel to Ireland occurred following the abolition of value added tax in January 1993 on imported second‐hand boats from the European Union (UK and continental Europe). This, together with a favourable monetary exchange rate at that time, may have increased the risk of invasion of the zebra mussel.     

Distribution of common octopus and common squid paralarvae in a wind-driven upwelling area (Ria of Vigo, northwestern Spain)

The variation of paralarval abundance in a region subjectedto wind-driven upwelling (Ria of Vigo, northwestern Spain) wasstudied. Research cruises were undertaken during the favourableupwelling season (May to October) in 2000 and 2001. Each cruiseincluded biological and hydrographic sampling and consistedof five stations in waters located to the east and west of theCies Islands. A total of 221 paralarvae of Octopodidae and Loliginidaewere collected over the 12-month study period. During 2000,higher abundance of paralarvae was observed in July, Septemberand October for Loligo vulgaris and Octopus vulgaris. In 2001,abundance of both species was higher in May and also in Septemberfor O. vulgaris. The mantle length of the paralarvae variedfrom 1.25 to 2.25 mm and from 1.00 to 4.90 mm within the O.vulgaris and L. vulgaris individuals, respectively. The presence/absenceof upwelling modulates the abundance and spatial distributionof loliginid and octopod paralarvae. The relationship betweenthe distribution and movement of these paralarvae in the Riaof Vigo seems to follow the circulation system defined for thisarea; when the upwelling extends its influence inside the Ria,the paralarvae are transported to the inner part in a west—eastdirection.     

Temperature requirements of Alnus pollen in Spain and Italy (1994-1998)

Woody plants in temperate regions, in order to prevent the water in their cells from freezing, interrupt their growth entering into a physiological state called "dormancy". Trees also have a heat requirement that must be fullfilled before actual growth is resumed and pollination occurs. The objective of this study is to ascertain the influence that a climatic parameter such as temperature exerts on Alnus, in order to identify the start of the dormancy period, its duration and the consequent heat requirement that triggers flowering in three European regions located at the same latitude. In this regard, we chose two areas in Spain (Vigo and Santiago de Compostela) and one in Italy (Perugia), since they have different temperature patterns, ranging from mild in the case of Vigo to cold in that of Perugia, including an intermediate temperature pattern (Santiago). Monitoring in the three stations was carried out by means of a 7-day Lanzoni VPPS 2000 pollen trap. Alnus began its pollen season between the second and third week of January in Spain, while in Italy it began during the first week of February. During the four years under study, in order to overcome the dormancy period, Perugia needed an average of 962 chilling hours (CH), Santiago 622 CH, and Vigo had the smallest chilling accumulation with only 460 CH. In the case of heat accumulation, we found that Perugia had the smallest requirement and Vigo the largest, thus Perugia needed an average of 42 (growth degree days) GDD, Santiago around 50 GDD and Vigo 62 GDD.     

After the fire: the end of a house life-cycle at the Iron Age site of Nabás (North-western Iberia)

Vegetation History and Archaeobotany - The existence of a fire event at the Iron Age hillfort of Nabás, which is located on the southern bank of the Ría de Vigo (Galicia, NW of the...     

If the Asian green mussel,Perna viridis (Linnaeus, 1758), poses the greatest invasive marine species threat to Australia,why has it not invaded?

A national approach has been developed to the problem of invasive marine species (IMS) in the Australian marine environment. Fifty-five species were listed as posing significant threats to Australia. A 2005 analysis of the scientific literature concluded that the Asian green mussel Perna viridis (Linnaeus, 1758) poses the greatest threat to Australia. The mussel has in fact successfully invaded many areas of the world’s oceans. Despite the numerous and varied opportunities for P. viridis to be distributed to northern Australia it has not established a known population on the continent, perhaps suggesting there are biological factors inhibiting its establishment. The invasion success of P. viridis in many parts of the world and its failure so far to establish in Australia make the species ideal for testing theories of the factors determining invasion success. Such research will allow a reconsideration of the invasion threat the species poses to the Australian marine environment.     

Differences in atmospheric trees pollen seasons in winter, spring and summer in two European geographic areas, Spain and Italy

The global climate change reported over recent years may prompt changes in the atmospheric pollen season (APS). The aim of this study is to evaluate the possible impact provoked by meteorological conditions variations at different seasons of the year or different geographical areas on APS. Alnus, Betula and Castanea atmospheric pollen seasons and trends during the last 17 years at Ourense and Vigo (Galicia—NW Spain) and Perugia (Italy) were analysed. Possible incidence of the meteorological trends observed in the different cities on the atmospheric pollen seasons and the chill and heat requirements were evaluated. Pollen data from Ourense, Vigo and Perugia (1995–2011) were used. Pollen sampling was performed using LANZONI VPPS 2000 volumetric traps (Hirst in Ann Appl Biol 36:257–265, 1952), placed on top of different buildings at a similar height from the ground. Several methods, dates and threshold temperatures for determining the chill and heat requirements needed to trigger flowering were tested. Different temporary order in the pollination sequence was observed between the three pollen types studied in the three sites. Alnus flowers few days in advance in Ourense respecting to Vigo and 1 month earlier than Perugia. The Betula flowering start date in Ourense and Vigo is almost simultaneous, taking place only 5 days in advance with respect to Perugia. Finally, scarce differences in the APS onset of Castanea were detected between the three cities. The variations observed among the two areas (Umbria, Italy and Galicia, Spain) in the onset of pollen season in the winter or spring flowering trees could be explained by differences in the thermal requirements needed for flowering as consequence of the climatic conditions recorded during the previous period to flowering. The length of the chilling and heat period as well as the thermal requirements obtained showed differences between geographical areas. The chill requirements accumulated were higher in Perugia than Ourense and Vigo. By contrary, the lowest heat accumulation was achieved in Perugia. The observed trends in the APS characteristics and the weather-related parameters were not homogeneous both in the pollen types and sites. The pollen index of Betula and Castanea pollen in Ourense shows a significant trend to increase.     

Tracing the late Holocene evolution of the NW Iberian upwelling system

This work analyses data from the OMEX core KSGX 40 (164-cm long) collected in the Galicia Mud Deposit, from the NW Iberian outer continental shelf, off the Ria de Vigo (North Spain). Sediment grain-size and benthic foraminifera distribution patterns indicate a number of substantial changes in sedimentation and also food/oxygen availability to the benthic ecosystem during the last ∼4.8 ka cal BP. Cluster analysis (Q-mode and R-mode) based on the most abundant taxa was used to recognize similarities within the data set. Three main clusters were established by Q-mode cluster analysis. The species distribution pattern of two of these three clusters is clearly related with changes in texture of the sediments. Textural data and benthic foraminifera proxies suggest that between ∼4.8 and 2.2 ka cal BP there was a period of higher hydrodynamism, with more energetic oceanic currents associated with shelf mixed waters. The period since ∼2.2 ka cal BP until present was characterized by generally lower bottom energetic conditions and by a supply of finer sediment, which was richer in organic matter leading to more eutrophic conditions in the benthic environment, mainly between ∼2.2 and 1.2 ka cal BP and since ∼0.5 ka cal BP until the present. Eutrophication was probably determined by strong water column stratified conditions and by coastal upwelling-dominated depositional intervals and/or by higher lateral flux of organic carbon. Since the upwelling increasing periods are in general due the intensification of northerly winds, both most productive upwelling-dominated periods may be attributed to climatic/oceanographic induced changes during the Sub-Atlantic climate. Small sea levels oscillations also should contribute to these results. Eutrophication since the end of Little Ice Age may be related to human influence in coastal areas.     

Genetic variation among populations of the endangered fan mussel <Emphasis Type="Italic">Pinna nobilis</Emphasis> (Mollusca: Bivalvia) along the Tunisian coastline

The Mediterranean fan mussel Pinna nobilis is an endangered invertebrate due to overexploitation and habitat deterioration. In this species, two distinctive morphs, the “combed” and “straight and wide” forms, have been recorded and it is not known whether this morphometric variability is attributed to genetic or environmental factors. In this study, we used mitochondrial COI sequences to describe the genetic variability of five Pinna nobilis populations sampled along the northern, eastern, and southern Tunisian coastline, and to examine whether there is a genetic differentiation between the two forms of this species. DNA sequences of 675 bp from the mitochondrial COI gene have revealed 10 different haplotypes among 49 examined specimens. Haplotype diversity was high, ranging between 0.40441 and 0.80952, and showed a decreasing North-East gradient, which seems to be explained by the hydrogeography of the study area. The mitochondrial COI marker did not suggest a genetic distinctiveness between the two Pinna nobilis shell forms, “combed” and “straight and wide”. Although the morphometric plasticity of the fan mussel could be due to the influence of environmental factors, further genetic studies using nuclear markers are envisaged to investigate whether this differentiation is associated to gene flow restrictions.     

Nuclear and mitochondrial genetic variability of an old invader, Dreissena polymorpha (Bivalvia), in French river basins

Analyzing non-native species after their establishment can shed light on factors responsible for their colonization success. The zebra mussel (Dreissena polymorpha), a species of Ponto-Caspian origin, is a worldwide invader, and was first reported in northern France in the nineteenth century. The genetic diversity and structure of D. polymorpha from five river basins (nine locations) in France were estimated using nuclear markers (amplified fragment length polymorphisms, AFLPs; 154 loci) and sequences from a 514 bp mitochondrial DNA region (cytochrome c oxidase subunit I; COI). Results from the two types of markers were generally congruent, although COI sequences showed limited genetic polymorphism, with one predominant and cosmopolitan haplotype (84 % of the individuals). Both COI and AFLPs revealed that the most divergent and the least polymorphic sample was the one from southern France. This population was the most distant geographically from all the others and among the most recent introductions reported in France. In addition, in northern France, AFLP loci revealed a significant river basin effect, despite a common pathway of introduction. This result suggests a shift in local population dynamics, perhaps due to stochastic recruitment, and underlines the importance of local dispersal (e.g. via planktonic larvae) compared to overland transport in shaping the population structure of this naturalized freshwater mollusk.     

Connectivity of populations within and between major biogeographic regions of the tropical Pacific in <Emphasis Type="Italic">Conus ebraeus</Emphasis>, a widespread marine gastropod

Information on genetic connectivity and structure of populations in the tropical Pacific is critical for making inferences about the origins and maintenance of diversity in this region. Sequences of the mitochondrial COI gene from 92 individuals of the trans-Pacific gastropod Conus ebraeus from eight localities spanning the tropical Pacific were analyzed to determine whether populations in the western, central, and eastern Pacific exhibit genetic structure, to examine the demographic histories of populations, and to infer patterns of gene flow. A total of 43 unique haplotypes were recovered, including a common haplotype that occurred in six of the eight populations examined. AMOVA and pairwise F-statistics showed that populations in the western and central Pacific were significantly differentiated from populations in the eastern Pacific, but no other evidence of structure. Bayesian isolation–migration (IM) analysis suggested that populations in the western and central Pacific separated from those in eastern Pacific during the Pleistocene. Examination of mismatch distributions and results from IM revealed that populations in the western and central Pacific expanded during the Pleistocene. Gene flow across the East Pacific Barrier appears to occur predominantly westward.