Epibiont assemblages on limpet shells: Biodiversity drivers in intertidal rocky shores.

Limpet shells can harbour a high diversity of species. Individuals of four limpet species (Patella depressa, Patella ulyssiponensis, Patella vulgata and Siphonaria pectinata) were collected monthly during one year in southern Portugal. Epibiont organisms were identified, counted and the percentage cover of facilitator taxa was also recorded. A total of 86 taxa were identified with abundance reaching 674 epibionts on a single basibiont shell. P. ulyssiponensis showed the highest epibiont species diversity and richness while P. depressa and P. vulgata showed similar diversity and richness. P. depressa had a more even epibiotic community mainly due to higher densities of Chthamalus sp. Overall, basibiont species was the key factor determining the epibiotic community, followed by month/season and erect algae. The presence of erect algae potentiated the epibionts diversity on limpet shells, whereas the occurrence of barnacles tended to decrease it and the presence of crustose algae had no significant effect on epibionts diversity. These findings shed further light on the biological and ecological complex relationships among keystone species inhabiting intertidal rocky shores.

Identification of Ensis siliqua samples and establishment of the catch area using a species-specific microsatellite marker.

European Council Regulation 104/2000 states that fishery products must be labeled to indicate commercial designation of species, the production method, and the catch area. Therefore, traceability of seafood implies knowledge of the species offered to retail and their origin. Ensis siliqua is a bivalve intensively fished in Europe and sold in fresh and canned forms. Although several published methods clearly differentiate Ensis genus species, none of those assess the origin of the commercial samples. In the present study, a microsatellite marker (Esi-UDC3055F) was developed to establish the catch area of E. siliqua samples. Amplification yielded a fragment of 275 or 302 base pairs, depending on whether they were Iberian or Irish populations. The usefulness of this method was also assessed in commercial samples. The results of this study provide a reliable methodology for the identification of catch area in European E. siliqua commercial samples. The coupling of this methodology with existing techniques for razor clam species identification provides a powerful tool for traceability and labeling enforcement.

Isolation of microsatellite markers and analysis of genetic diversity among east Atlantic populations of the sword razor shell Ensis siliqua: a tool for population management.

The sword razor shell Ensis siliqua (Linnaeus, 1758) is found mainly from Norway to the Atlantic coast of the Iberian Peninsula. It is intensively caught in Europe, being highly appreciated as seafood. To help in its conservation and management, five microsatellite markers were isolated and genetic variation was analyzed in samples from Ireland, Spain, and Portugal. The highly significant differentiation (θ = 0.287, P < 0.001) observed was mainly due to differences between samples from Irish and Iberian Peninsula localities, except Aveiro (its sample resembled the Irish samples, and it may be predominantly self-recruiting). These groups of samples showed significant differences in allelic richness that could be related to harvesting intensity. Moreover, microsatellites detected low but significant differentiation between Iberian localities (Celeiro and Olhão), and Aveiro differed significantly from Strangford Lough. Overall, results suggest that two independently evolving regions exist and that management strategies should be designed for each region.

Environmental quality assessment combining sediment metal levels, biomarkers and macrobenthic communities: application to the Óbidos coastal lagoon (Portugal).

Macroinvertebrate benthic communities are one of the key biological components considered for the assessment of benthic integrity in the context of the Water Framework Directive (WFD). However, under moderate contamination scenarios, the assessment of macrobenthic alterations at community level alone could be insufficient to discriminate the environmental quality of coastal and transitional waters. Keeping this in view, sediment quality of moderately contaminated sites in a coastal lagoon (Óbidos lagoon, Portugal) was assessed by the combination of sediment metal levels, Carcinus maenas biomarkers (accumulated metals and oxidative stress responses) and macrobenthic communities. Two sites were selected in confined inner branches (BS and BB) and a third one in the middle lagoon (ML). The site BB presented slightly higher levels of metals in sediment but biological variables calculated for macrobenthic data were not significantly different between sites. The biotic index M-AMBI that is being applied to assess environmental quality of transitional waters in the scope of the WFD pointed either to high (site ML) or good quality status (BS and BB) in the selected sites. However, crabs from BB site presented significantly higher levels of Ni in hepatopancreas than those from ML and macrobenthic community structure was significantly different between BB and ML. Additionally, spatial differences were obtained for oxidative stress parameters suggesting that BB site presented stressors for crabs (higher GST and lower GSH(t) at BB site). Factor analysis (PCA) integrating sediment contamination, biomarkers in crabs and macrobenthic data also distinguished BB site as the most environmentally disturbed. On the other hand, at ML site, some macrobenthic variables (equitability and polychaetes' diversity) were found to be enhanced by current environmental conditions, suggesting the existence of a better sediment quality. Current results pointed to the usefulness of integrating macrobenthic community alterations with responses at organism level (bioaccumulation and biochemical endpoints) in order to increase the accuracy of environmental quality assessment in lagoon systems. Moreover, the application of different statistical methods was also found to be recommendable.

Combined use of Nassarius reticulatus imposex and statolith age determination for tracking temporal evolution of TBT pollution in the NW Portuguese continental shelf.

Spatial and temporal trends of tributyltin (TBT) pollution on the northern continental shelf of Portugal (5-34 m depth) were assessed using Nassarius reticulatus as a bioindicator. Imposex levels were determined in 2006 and 2010, and integrated with age readings from statoliths for a better chronological analysis of the data. Females affected with imposex were detected in 84% of the sites surveyed in 2006, with levels higher than the Ecological Quality Objective (EcoQO) set by the OSPAR Commission for N. reticulatus (VDSI < 0.3) in approximately half of those sites. In 2010, imposex was only detected in 20% of the surveyed sites and, when detected, the VDSI were lower than the EcoQO, except for one location. The comparison between the current results and the levels previously reported for 2004 and 2005 in the same offshore area evidences a clear decline of imposex in 2010 that confirms the efficacy of Regulation (EC) No. 782/2003 and the IMO AFS-Convention (entered into force in 2003 and 2008, respectively). Moreover, the age of the whelks used in the monitoring is estimated in 3-5 years, indicating that actual TBT water contamination in 2010 may in fact be lower than that extrapolated from the imposex levels. We conclude that the legislation was indeed very effective in reducing TBT pollution in the NW Portuguese continental shelf, not only in terms of the magnitude but also in terms of fast field repercussion.

Factors structuring temporal and spatial dynamics of macrobenthic communities in a eutrophic coastal lagoon (Óbidos lagoon, Portugal).

The present work aimed to identify the main environmental drivers shaping temporal and spatial dynamics of macrobenthic communities within a eutrophic coastal lagoon. Sediments in the Óbidos lagoon showed a gradient of increasing metal contamination from the inlet area to inner branches. The mid-lower lagoon area exhibited an intermediate contaminated condition between the inlet and upstream areas, suggesting that the effects of the organic loadings into the lagoon may be reflected until this area. This transitional feature was corroborated by biological data, with macrobenthic assemblages displaying characteristics of down- and upstream areas. Macrobenthic abundance peaked in winter, which was associated with a higher nutrient availability (mainly ammonium) and the proliferation of green macroalgae in mid-lower and inner lagoon areas. However, massive macroalgae growth resulted in a sharp decrease of macrobenthic diversity and abundance in spring, particularly where the higher amounts of decaying algae were detected. Higher dissimilarities between assemblages were detected during winter (and spring, for trophic composition), while in summer, differences were highly attenuated. The least contaminated area (close to the sea inlet) experienced smaller temporal variations for environmental variables, as well as the lowest temporal biological variability. This area was dominated by carnivores, which were related with increased salinity. Deposit-feeders were numerically dominant in the lagoon, being generally spread within organically enriched sandy and muddy areas. The high concentration of chlorophyll a and suspended particulate matter in water was reflected in the abundance of deposit-feeders/suspension-feeders, taking benefit of the high primary productivity. On the other hand, deposit-feeders/herbivores responded to the decay of macroalgae mats in the sediment. Biological associations varied with the biological data used (taxonomic versus trophic group composition; abundance versus biomass), highlighting the relevance of the combination of different data analysis' approaches. In general, BIOENV analysis indicated total phosphorus, biomass of Ulva, metals and organic carbon and nitrogen as being significantly influencing benthic patterns. On the other hand, discrepancies in ecological behaviours of some taxa were also detected in the present study stressing the need for additional studies on the relationships between macrobenthic communities and environmental variables. Implications of the present results for monitoring studies are discussed.

Inshore/offshore gradients of imposex and organotin contamination in Nassarius reticulatus (L.) along the Portuguese coast.

Imposex and organotin (OT) tissue contamination of the netted whelk Nassarius reticulatus (L.) were assessed in the continental shelves around the main estuaries of the central coast of Portugal (Lisbon: Tagus estuary; Setúbal: Sado estuary) and the main coastal lagoon in the Southern of Portugal (Faro: Ria Formosa). Pollution levels were higher in areas of more intense boat traffic and shipyard activities and imposex showed a clear decreasing gradient from the estuaries to the offshore, in relation to a similar gradient of tissue contamination by tributyltin. Remarkably, imposex was extensively spread over the adjacent continental shelves of Tagus and Sado estuaries. The current work shows that TBT pollution is undoubtedly a matter of concern not only for the above estuaries where harbours are implanted but also for the adjacent continental shelves, regardless the massive dilution of contaminants that may occur in these deeper areas.

Doubly uniparental inheritance (DUI) of mitochondrial DNA in Donax trunculus (Bivalvia: Donacidae) and the problem of its sporadic detection in Bivalvia.

Mitochondrial DNA is transmitted maternally in metazoan species. This rule does not hold in several species of bivalves that have two mtDNA types, one that is transmitted maternally and the other paternally. This system of mitochondrial DNA transmission is known as doubly uniparental inheritance (DUI). Here we present evidence of DUI in the clam Donax trunculus making Donacidae the sixth bivalve family in which the phenomenon has been found. In addition, we present the taxonomic affiliation of all species in which DUI is currently known to occur and construct a phylogeny of the maternal and paternal genomes of these species. We use this information to address the question of a single or multiple origins of DUI and to discuss whether failed attempts to demonstrate the presence of DUI in several bivalve species might be due to problems of detection or to genuine absence of the phenomenon.

Use of shape to distinguish Chamelea gallina and Chamelea striatula (Bivalvia: Veneridae): Linear and geometric morphometric methods.

The commercially fished striped venus clams Chamelea gallina and C. striatula (Bivalvia: Veneridae) are difficult to distinguish by inexperienced observers and the taxonomy of these species is still an issue of discussion. The differences in shape between C. gallina and C. striatula from Portuguese coastal waters were studied through conventional linear and geometric morphometric analysis, using both contour (elliptic Fourier analysis) and landmark-based methods. The relationships shell length vs. height, width, and total weight were significantly different between species. However, because there was a considerable overlap in the data sets, the species could not be distinguished using any combination of those linear measurements. Geometric morphometric methods provided shape variables that led to 0-6% misclassification rates between species; linear morphometric measures led to 16.8% error. Contour analysis revealed differences primarily in the shell umbo and lunular area. The umbo was more "sharp" and the lunula less pronounced in C. striatula than in C. gallina. Generalized procrustes superimposition (landmark analysis) showed that the main differences between species reside in the length of the pallial sinus. Thus, an index was developed (PI: Pallial Index = pallial sinus length/shell length), which was successfully used to separate the species (with 100% correct classification), i.e., specimens with PI lower than 0.119 belonged to C. gallina, whereas greater PI values were attributed to C. striatula. The use of these geometric morphometric methods allowed the detection of differences in shape between these two species and to develop an easy-to-use identification index. We encourage the development of analogous indices that apply the methods of geometric morphometrics to distinguish between other species whose identification is complicated.

Distribution patterns of macrobenthic species in relation to organic enrichment within aquaculture earthen ponds.

The relationship between organic enrichment and macrobenthic colonization patterns was investigated during an 8-month period in Diplodus sargus (white seabream) production ponds. A stratified sampling design was applied and each pond was divided into three zones: water entrance (WE); central (C); and automatic feeder zones (AF). Generally, the number of species and Shannon-Wiener diversity increased from the WE to the AF zone. Abundance did not present a clear trend. The recently developed marine biotic index (AMBI) was applied and showed to be sufficiently robust to discriminate, within a relatively small area, differences in macrobenthic communities due to organic enrichment. Nevertheless, caution is advised when applying this index or others based on ecological group's assignment, as the classification of a certain area may differ when allocating a certain species to an unsuitable group. This is particularly evident when common species are involved.

The use of the marine biotic index AMBI in the assessment of the ecological status of the Obidos lagoon (Portugal).

This study reports the longitudinal distribution of sediment properties, including inorganic and organic contaminants, and the structure of the benthic community in Obidos lagoon, a coastal system permanently connected to the sea and with negligible freshwater sources. Sediments from the upper to central lagoon consist of fine particles (91%) and from the lower lagoon of sands (94%). Chemical composition is strongly correlated to the percentage of fine particles. Contamination is relatively low in those sediments suggesting the effect of diffuse sources. The increase in organic matter content from down- to upstream areas was associated with the dominance of opportunistic species, while sensitive and indifferent species to organic enrichment were mainly associated to the clean sandy downstream area. The marine biotic index (AMBI) was suitable for the discrimination of the biological and environmental gradients in the Obidos lagoon and was highly related with the gradient of organic matter content in this system.