Nathalie Caill-Milly

Shell Shape Analysis and Spatial Allometry Patterns of Manila Clam (Ruditapes philippinarum) in a Mesotidal Coastal Lagoon

While gradual allometric changes of shells are intrinsically driven by genotype, morphometrical shifts can also be modulated by local environmental conditions. Consequently the common use of a unique dimension (usually length) to assess bivalves’ growth may mask phenotypic differences in valve shape among populations. A morphometric exhaustive study was conducted on Manila clam, Ruditapes philippinarum, by acquiring data in the French Arcachon Bay (intrasite phenotypic variability) and by comparing with other sites in the literature (intersite phenotypic variability). 2070 shells were subsampled, weighted, and automatically measured using TNPC software. Some ratios’ values indicate a relatively round and globular shape shell in comparison with other sites confirming poor conditions for some individuals. Among adult clams, three main morphological groups were identified and discussed according to spatial considerations. Allometric relations for pairs of shell descriptors were determined by testing classical linear and piecewise regression models on log-transformed relation of Huxley. A significant shape change correlated to size was observed; it corresponds to the second year of life of the clam. Relationships between density, disease, and shell shape are demonstrated and discussed related to other potential factors affecting shell shape. Finally, consequences on population regulation are addressed.

Brown muscle disease (BMD), an emergent pathology affecting Manila clam Ruditapes philippinarum in Arcachon Bay (SW France)

We describe an emerging pathology, brown muscle disease (BMD), which specifically affects the Manila clam Ruditapes philippinarum in Arcachon Bay (France). BMD induces a transformation of the posterior adductor muscle, which becomes infused by conchiolin and calcified, reducing the ability of clams to bury. The disease affects both types of muscular tissue, with striated muscle becoming affected to a higher degree than smooth muscle. Two indices were created to quantify the symptoms: the Muscle Print Index, used for empty and live shells, and the Final Disease Index, utilized for live clams only. Histological sections were made and observed under light microscopy to examine the muscular damage and to investigate a causal agent. Sections revealed an important inflammatory response with a large invasion of hemocytes into tissues and a heavy necrosis of muscular fibers. Additionally, molecular biology analyses were carried out to search for bacteria and protozoan agents using generic primers. In both histological and molecular assays, bacteria and protozoans were discounted. We monitored 4 sites scattered around the bay over 2 yr. The mean prevalence was <12% without seasonal variation in 3 sites against 30% and a winter peak in 1 site. The latter site was accurately surveyed and revealed that clams at the sediment surface (abnormal position) were affected 3 times more frequently than buried clams (normal position).

Virus-like particles associated with brown muscle disease in Manila clam, Ruditapes philippinarum, in Arcachon Bay (France)

Recently, Manila clam, Ruditapes philippinarum, populations have suffered mortalities in Arcachon Bay (SW France). Mortality was associated with extensive lesions of the posterior adductor muscle, which become progressively brown and calcified. Ultrastructural observations by transmission electron microscopy revealed tissue degradation with necrotized muscle fibres and granulocytomas. Unenveloped virus-like particles (VLPs) were detected in muscle, granulocytic, epithelial and rectal cells. VLPs were abundant in the extracellular space, in the cytoplasm (free or enclosed in vesicles) and in the nucleoplasm of granulocytes. Nuclei and mitochondria of granulocytes displayed changes which suggested reactive oxygen species production and apoptosis induction. VLPs exhibited an icosahedral structure with a diameter of 25 to 35 nm. These observations suggest that the VLPs could belong to the family Picornaviridae or the Parvoviridae.

Heavy metals and organochlorinated compounds in the European eel (Anguilla anguilla) from the Adour estuary and associated wetlands (France)

Heavy metals and organic pollutants were investigated in the Adour estuary (South West France) and associated wetlands using the European eel (Anguilla anguilla) as a bioindicator. Heavy metals (Cu, Cd, Zn, Pb, and Ag) were measured in soft tissue of yellow eels. Mercury (total Hg and MeHg) and organochlorinated compounds (7 PCBs, 11 OCPs) were analysed in muscle. Concentrations in muscle were in agreement with moderately contaminated environments in Europe and were below the norms fixed for eel consumption for heavy metals and OCPs. Analyses of liver showed a higher pressure of Ag and Zn in the downstream estuary than in the freshwater sites whereas Cd was lower in the estuary probably because of the salinity influence. According to quality classes 100% of eels from freshwater sites indicated clean or slightly polluted environments. However, total mercury concentrations were close to the thresholds fixed by the European Community in the downstream estuary, whereas the sum of PCBs was found to be greatly above the fixed value. 100% of the individuals from the estuary were classified in quality classes corresponding to polluted or highly polluted sites. These first results highlight the need of further investigations focused on mercury and PCBs in this area taking the seasonal temperature influence into account for a better understanding of the pollution distribution and the possible threat on the eel population from the Adour basin.

Correlation between perkinsosis and growth in clams Ruditapes spp.

Perkinsosis is one of the most widespread diseases affecting commercially important species of molluscs globally. We examined the impact of Perkinsus spp. on shell growth at the individual scale in 2 clam species: Ruditapes decussatus from Mundaka Estuary (Spain) and R. philippinarum from Arcachon Bay (France). At Arcachon, 2 contrasting sites in terms of environment and Perkinsus olseni presence were chosen: Arguin (disease-free) and Ile aux Oiseaux (infected site). We monitored the dynamics of perkinsosis over the course of the experiment at Mundaka and Ile aux Oiseaux. Prevalences were high (>70%), and intensities were around 105 cells g-1 wet gills at Ile aux Oiseaux, and 106 cells g-1 at Mundaka. No significant differences in prevalence or intensity were observed over time. A 2 yr field growth experiment of tagged-recaptured clams was performed to determine individual clam growth rate, condition index (CI), and Perkinsus spp. infection intensity. Clams were collected at Ile aux Oiseaux and transplanted to Arguin. The growth rate was always significantly and negatively correlated with Perkinsus spp. infection, and positively correlated with CI. CI and Perkinsus spp. infection explained 19% and 7% of the variability of the growth rate at Mundaka and Ile aux Oiseaux, respectively. In experimental clams at Arguin, P. olseni infection explained 26% of the variability of the growth rate at the lower tidal level. Our results suggest that at a concentration of between 105 and 106 cells g-1, perkinsosis affects the physiological functions of the clams, highlighted by its impact on the growth rate.

Tidal level influence on the spawning process of the sea urchin Paracentrotus lividus (Lamarck, 1816) on a rocky shore (Bay of Biscay)

The sea urchin Paracentrotus lividus is an important exploited resource on the Mediterranean coast and has been the subject of much research. In the Bay of Biscay, the situation is different, as some studies have been conducted on the Spanish Basque coast but few on the French part of that coast. However, the Basque coast offers favourable conditions for the development of this species, and its exploitation could be a source of potential diversification for fishermen, especially in the context of difficulties with some other resources. At the request of the managers of this coastal resource, a study was undertaken on sea urchins to improve knowledge of this stock, particularly about its biological characteristics. In this work, assessment consisted of determining the spawning process of the stock throughout the year by collecting data on sea urchin size and wet weights of the test, dry gonads and viscera. From September 2013 to September 2014, sea urchins were collected monthly at this site, from intertidal and subtidal areas, and then analysed in the laboratory. This study takes into account the tidal level in the sample design, which allows a novel highlighting of significant differences in biological characteristics between intertidal and subtidal individuals. Spawning occurs mainly during May and June, which is important knowledge for defining relevant management measures for fisheries.

Absence of spatial genetic structure in common dentex (Dentex dentex Linnaeus, 1758) in the Mediterranean Sea as evidenced by nuclear and mitochondrial molecular markers

The common dentex, Dentex dentex, is a fish species which inhabits marine environments in the Mediterranean and Northeast Atlantic regions. This is an important species from an ecological, economic and conservation perspective, however critical information on its population genetic structure is lacking. Most samples were obtained from the Mediterranean Sea (17 sites) with an emphasis around Corsica (5 sites), plus one Atlantic Ocean site. This provided an opportunity to examine genetic structuring at local and broader scales to provide science based data for the management of fishing stocks in the region. Two mitochondrial regions were examined (D-loop and COI) along with eight microsatellite loci. The COI data was combined with publicly available sequences and demonstrated past misidentification of common dentex. All markers indicated the absence of population genetic structure from the Bay of Biscay to the eastern Mediterranean Sea. Bayesian approaches, as well as the statistical tests performed on the allelic frequencies from microsatellite loci, indicated low differentiation between samples; there was only a slight (p = 0.05) indication of isolation by distance. Common dentex is a marine fish species with a unique panmictic population in the Mediterranean and likely in the Atlantic Ocean as well.