Influence of sea-ice dynamics on coastal Antarctic benthos: A case study on lantern clams (Laternula elliptica) in Adélie Land.

Polar regions are warming faster than the world average and are profoundly affected by changes in the spatio-temporal dynamics of sea ice, with largely unknown repercussions on the functioning of marine ecosystems. Here, we investigated the impacts of interannual sea-ice variability on coastal benthic communities in Antarctica, focusing on a close-to-pristine area (Adélie Land). We investigated shell growth of the circum-Antarctic bivalve Laternula elliptica, considered a key species in these soft bottom benthic communities. Chondrophores of live-collected clams were prepared using standard sclerochronological methods to study the interannual variability of shell growth from 1996 to 2015. Our results show that the master chronology varied with sea-ice dynamics. When sea ice breaks up too early, sympagic algae do not have time to accumulate sufficiently high biomass, thus strongly limiting the energy input to the benthos. This negatively affects the physiological performance of L. elliptica, thereby altering their population dynamics and hence the functioning of these soft-bottom ecosystems.

Marine Pollutant Tributyltin Affects DNA Methylation and Fitness of Banded Murex (Hexaplex trunculus) Populations.

Banded murex, Hexaplex trunculus, is a marine gastropod whose reproductive fitness can be severely affected by very low concentrations of antifouling compound tributyltin (TBT). TBT has strong xenoandrogen impacts on snails, causing the development of imposex (e.g., the superimposition of male sexual characteristic in females), thereby affecting the fitness of entire populations. TBT is also known as a DNA-demethylating agent and an obesogenic factor. The aim of this study was to unravel the interactions between TBT bioaccumulation, phenotypic responses, and epigenetic and genetic endpoints in native populations of H. trunculus. Seven populations inhabiting environments along the pollution gradient were sampled in the coastal eastern Adriatic. These included sites of intense marine traffic and boat maintenance activity and sites with low anthropogenic impact. Populations inhabiting intermediately and highly polluted sites exhibited higher TBT burdens, higher incidences of imposex, and higher wet masses of snails than populations in lowly polluted sites. Other morphometric traits and cellular biomarker responses did not show clear differentiation among populations in relation to marine traffic/pollution intensity. An analysis of methylation sensitive amplification polymorphism (MSAP) revealed environmentally driven population differentiation and higher epigenetics than genetic within-population diversity. Moreover, decreases in genome-wide DNA methylation coincided with the imposex level and snail mass, suggesting an epigenetic background of the animal phenotypic response.

Otolith fingerprints reveals potential pollution exposure of newly settled juvenile Sparus aurata.

Coastal ecosystems are increasingly threatened by a wide range of human activities. Fish otolith chemistry, by creating a unique specific signature, can be used as a natural tag for determining life stage dispersal, spatial connectivity and population structure. In this study, we tested whether differences in otolith composition among juveniles of gilthead sea bream, Sparus aurata, could enable their proper allocation to polluted areas based on higher concentrations of elements related to contaminants. Otoliths were embedded, sectioned and analysed by LA-ICP-MS in line scan mode. Multivariate analysis confirmed clear separation between sites and elements. Samples from the site under the strongest anthropogenic impact from industrial and agricultural river input were characterized by higher values of Pb/Ca and Zn/Ca. However, these relatively low values likely do not have a negative effect on S. aurata recruitment, though they could serve for identifying the contribution of polluted nurseries to stock dynamics.

Competitive feeding interactions between native Ostrea edulis and non-native Crassostrea gigas with implications of introducing C. gigas into commercial aquaculture in the eastern Adriatic Sea.

In order to detect the possible regulatory effect of non-native C. gigas on the native O. edulis, under aquaculture conditions, feeding interactions between them were investigated in a highly productive environment of Lim Bay (Adriatic Sea). The present study uses a multi-methodological approach, including stomach content, DNA barcoding and stable isotope analysis to elucidate the feeding ecology of two oyster species. The research confirmed a high overlap throughout the year in the feeding traits among native and non-native oyster species. Competition for food was not the only relationship that exists between the investigated species as the presence of O. edulis larvae in C. gigas stomach content was confirmed by DNA analysis. Findings are not in favour of introducing C. gigas to commercial aquaculture in any new areas in the Adriatic Sea and support the need to improve the existing O. edulis aquaculture and conserve its wild stocks.

A network of bivalve chronologies from semi-enclosed seas.

Four chronologies of the bivalve species Glycymeris pilosa have been constructed along a 300 km gradient of the eastern coastal Adriatic Sea, all of which span the common period of 1982-2015. The chronologies are compared to local and remote environmental drivers suspected to influence the biology of the system, including air and seawater temperature, precipitation and freshwater discharge. The Adriatic-Ionian Bimodal Oscillating System (BiOS), a key oceanographic feature quantified by satellite-derived absolute dynamic topography, is also compared to the chronologies. The chronologies at the two southern sites are more strongly influenced by local river discharge, while the two northern chronologies are more strongly influenced by BiOS. These results highlight the broadscale importance of BiOS to the Adriatic system as well as the heterogeneity of nearshore environmental and drivers of growth. These G. pilosa chronologies provide unique multidecadal, continuous, biological time series to better understand the ecology and fine-scale variability of the Adriatic with potential for other shallow, semi-enclosed seas.

Glycymeris pilosa (Bivalvia) - A high-potential geochemical archive of the environmental variability in the Adriatic Sea.

Due to its outstanding longevity (decades), the shallow-water bivalve Glycmeris pilosa represents a prime target for sclerochronological research in the Mediterranean Sea. In the present study, we analyzed the microgrowth patterns and the stable carbon (δ13Cshell) and oxygen (δ18Oshell) isotopes of the outer shell layer of live-collected G. pilosa specimens from four different sites along the Croatian coast, middle Adriatic Sea. Combined analysis of shell growth patterns and temporally aligned δ18Oshell data indicated that the main growing season lasts from April to December, with fastest growth rates occurring during July and August when seawater temperatures exceeded 22 °C. Slow growth in the cold season (<12 °C) coincided with the formation of winter growth lines on the outer shell surface. The growth cessation occurred in winter, but on the outer shell surface the brown summer bands are more pronounced than the winter lines. Mutvei-staining of cross-sections facilitated the recognition of the growth lines. δ13Cshell values reflect ontogenetic changes in physiology as well as seasonal changes in primary production and salinity.

Bivalve trophic ecology in the Mediterranean: Spatio-temporal variations and feeding behavior.

The trophic ecology of two bivalves, the clam Callista chione and the cockle Glycymeris bimaculata was studied using environmental and biochemical variables of the suspended particulate matter and the sediment. Samples were collected from two shallow sites, Pag and Cetina, in the coastal oligotrophic Mediterranean Sea, during a 17 month period. The temporal variation of the particulate matter reflected a mixture between marine and terrestrial sources throughout the year, with a clear marine influence during summer and fall, and predominance of terrestrial inputs during spring and winter. The digestive gland was a useful rapid turnover tissue, where the carbon isotope signal was species-specific and the nitrogen isotope one was site-specific. FA markers in the digestive gland revealed a mixed diet where Callista chione fed more upon fresh material than G. bimaculata which relied largely on bacteria-derived detritus. Overall, little feeding niche overlap was observed between the two species during the year, indicating resource partitioning, expected for a food-limited system. The present trophic ecology study in co-occurring species allowed identifying species-specific feeding adaptations to environmental variability.

Historical baselines in marine bioinvasions: Implications for policy and management.

The human-mediated introduction of marine non-indigenous species is a centuries- if not millennia-old phenomenon, but was only recently acknowledged as a potent driver of change in the sea. We provide a synopsis of key historical milestones for marine bioinvasions, including timelines of (a) discovery and understanding of the invasion process, focusing on transfer mechanisms and outcomes, (b) methodologies used for detection and monitoring, (c) approaches to ecological impacts research, and (d) management and policy responses. Early (until the mid-1900s) marine bioinvasions were given little attention, and in a number of cases actively and routinely facilitated. Beginning in the second half of the 20th century, several conspicuous non-indigenous species outbreaks with strong environmental, economic, and public health impacts raised widespread concerns and initiated shifts in public and scientific perceptions. These high-profile invasions led to policy documents and strategies to reduce the introduction and spread of non-indigenous species, although with significant time lags and limited success and focused on only a subset of transfer mechanisms. Integrated, multi-vector management within an ecosystem-based marine management context is urgently needed to address the complex interactions of natural and human pressures that drive invasions in marine ecosystems.

Changes in the tissue concentrations of trace elements during the reproductive cycle of Noah's Ark shells (Arca noae Linnaeus, 1758).

Concentrations of 23 trace elements (TEs; essential: Co, Cu, Fe, Mn, Mo, Se, V, Zn; non-essential: Ag, Al, As, Ba, Cd, Cr, Cs, Li, Ni, Pb, Rb, Sr, Ti, Tl, U) in whole soft tissues of Noah's Ark shell (Arca noae) were determined monthly during one year (March 2013-February 2014) at two sampling sites in the central part of the Eastern Adriatic Sea. Our aim was to detect the influence of reproductive cycle and changes in the environmental factors on the variabilities of TEs' contents. Higher concentrations of Pb, Ba, V, Mo, Mn and Fe were found at potentially contaminated site in Pasman channel, whereas higher concentrations of Tl, Ni, Li, Cr, Cd, Ti and Se were found at reference site in Nature Park Telascica. Since several bioaccumulated TEs were associated to mean gonadal index, in TEs monitoring in A. noae, animal gonadal status has to be considered.

Drivers of shell growth of the bivalve, Callista chione (L. 1758) - Combined environmental and biological factors.

Seasonal shell growth patterns were analyzed using the stable oxygen and carbon isotope values of live-collected specimens of the bivalve Callista chione from two sites in the Adriatic Sea (Pag and Cetina, Croatia). Micromilling was performed on the shell surface of three shells per site and shell oxygen isotopes of the powder samples were measured. The timing and rate of seasonal shell growth was determined by aligning the δ18Oshell-derived temperatures so that the best fit was achieved with the instrumental temperature curve. According to the data, shells grew only at very low rates or not at all during the winter months, i.e., between January and March. Shell growth slowdown/shutdown temperatures varied among sites, i.e., 13.6 °C at Pag and 16.6 °C at Cetina, indicating that temperature was not the only driver of shell growth. Likely, seasonal differences in seawater temperature and food supply were the major component explaining contrasting growth rates of C. chione at two study sites. Decreasing shell growth rates were also associated with the onset of gametogenesis suggesting a major energy reallocation toward reproduction rather than growth. These results highlight the need to combine sclerochronological analyses with ecological studies to understand life history traits of bivalves as archives of environmental variables.

Influence of riverine input on the growth of Glycymeris glycymeris in the Bay of Brest, North-West France.

A crossdated, replicated, chronology of 114 years (1901-2014) was developed from internal growth increments in the shells of Glycymeris glycymeris samples collected monthly from the Bay of Brest, France. Bivalve sampling was undertaken between 2014 and 2015 using a dredge. In total 401 live specimens and 243 articulated paired valves from dead specimens were collected, of which 38 individuals were used to build the chronology. Chronology strength, assessed as the Expressed Population Signal, was above 0.7 throughout, falling below the generally accepted threshold of 0.85 before 1975 because of reduced sample depth. Significant positive correlations were identified between the shell growth and the annual averages of rainfall (1975-2008; r = 0.34) and inflow from the river Elorn (1989-2009; r = 0.60). A significant negative correlation was identified between shell growth and the annual average salinity (1998-2014; r = -0.62). Analysis of the monthly averages indicates that these correlations are associated with the winter months (November-February) preceding the G. glycymeris growth season suggesting that winter conditions predispose the benthic environment for later shell growth. Concentration of suspended particulate matter within the river in February is also positively correlated with shell growth, leading to the conclusion that food availability is also important to the growth of G. glycymeris in the Bay of Brest. With the addition of principle components analysis, we were able to determine that inflow from the River Elorn, nitrite levels and salinity were the fundamental drivers of G. glycymeris growth and that these environmental parameters were all linked.

Combined Use of Morphological and Molecular Tools to Resolve Species Mis-Identifications in the Bivalvia The Case of Glycymeris glycymeris and G. pilosa.

Morphological and molecular tools were combined to resolve the misidentification between Glycymeris glycymeris and Glycymeris pilosa from Atlantic and Mediterranean populations. The ambiguous literature on the taxonomic status of these species requires this confirmation as a baseline to studies on their ecology and sclerochronology. We used classical and landmark-based morphometric approaches and performed bivariate and multivariate analyses to test for shell character interactions at the individual and population level. Both approaches generated complementary information. The former showed the shell width to length ratio and the valve asymmetry to be the main discriminant characters between Atlantic and Mediterranean populations. Additionally, the external microsculpture of additional and finer secondary ribs in G. glycymeris discriminates it from G. pilosa. Likewise, landmark-based geometric morphometrics revealed a stronger opisthogyrate beak and prosodetic ligament in G. pilosa than G. glycymeris. Our Bayesian and maximum likelihood phylogenetic analyses based on COI and ITS2 genes identified that G. glycymeris and G. pilosa form two separate monophyletic clades with mean interspecific divergence of 11% and 0.9% for COI and ITS2, respectively. The congruent patterns of morphometric analysis together with mitochondrial and nuclear phylogenetic reconstructions indicated the separation of the two coexisting species. The intraspecific divergence occurred during the Eocene and accelerated during the late Pliocene and Pleistocene. Glycymeris pilosa showed a high level of genetic diversity, appearing as a more robust species whose tolerance of environmental conditions allowed its expansion throughout the Mediterranean.

The bivalve Glycymeris pilosa as a multidecadal environmental archive for the Adriatic and Mediterranean Seas.

We evaluated the potential of Glycymeris pilosa as an environmental indicator for the Mediterranean region by applying sclerochronological techniques on a sample set collected from Pasman Channel in the middle Adriatic Sea. Maximal longevity of analyzed shells was 69 years. Growth increments in acetate peels of the hinge region had clear boundaries, and there was a strongly synchronous signal in growth-increment width among individuals. The final, replicated chronology spanned 1969 to 2013. Shell growth negatively correlated with local summer sea temperatures and positively with November precipitation. High correlation between shell growth and circulation patterns in the northern Ionian was also observed, with slower growth occurring during cyclonic regimes. Given its broad distribution in the region and the ability to crossdate, generate annually-resolved chronologies, and of a length that substantially overlaps with observational records, G. pilosa has considerable potential to test hypotheses relating to environmental variability and biological response in the Mediterranean.

Endoliths in Lithophaga lithophaga shells--Variation in intensity of infestation and species occurrence.

Pronounced differences with respect to the extent of infestation and the degree of Lithophaga lithophaga shell damage inflicted by euendolithic taxa at two sites in the Adriatic Sea representing different productivity conditions, are described. Shells collected from the eastern part of Kastela Bay, which is characterized by higher primary productivity, have significantly more shell damage then the shell collected from a site on the outer coast of the island of Ciovo exposed to the oligotrophic Adriatic Sea. The presence of endoliths and their perforations were detected in different layers of the shell, including solidly mineralized parts of the skeleton and within the organic lamellae incorporated into the shell. Phototrophic endoliths were not observed in the specimens. The most serious damage to L. lithophaga shells was the boring clionaid sponge Pione vastifica, which was more common in shells collected from Kastela.

Reaction of the mussel Mytilus galloprovincialis (Bivalvia) to Eugymnanthea inquilina (Cnidaria) and Urastoma cyprinae (Turbellaria) concurrent infestation.

In total 480 individuals of Mytilus galloprovincialis were sampled monthly from October 2009 to September 2010, at the shellfish farm in the Mali Ston Bay, south Adriatic Sea (Croatia) in order to assess the extent of pathology imposed by two parasites, Eugymnanthea inquilina (Cnidaria) and Urastoma cyprinae (Turbellaria). Although a deteriorating impact on host reproduction or condition index was lacking, we evidenced ultrastructural and functional alteration in host cells at the attachment site. Ultrastructural changes included hemocytic encapsulation of the turbellarian and cell desquamation in medusoid infestation. Caspase positive reaction inferred by immunohistochemistry (IHC) was triggered in cases of turbellarian infestation, in contrast with hydroids, suggesting that the former exhibits more complex host-parasite interaction, reflected in the persistent attempts of the parasite to survive bivalve reaction. We have evidenced that both organisms trigger specific host reaction that although not costly in terms of host reproductive cycle or growth, results in mild tissue destruction and hemocyte activation. A lower degree of tissue reaction was observed in cases of hydroid infestation, compared to turbellarian.