Éditeur(s) : Actes du 6e colloque national BRG, La Rochelle, 2-3-4 octobre 2006
Résumé : Previous studies have shown heritable variation in larval developmental traits in the Pacific oyster Crassostrea gigas. In order to study the genetic consequences of production of oyster larvae in hatcheries, two factors, specific to hatcheries, were examined: the effect of discarding the smallest larvae (i.e. culling) and the effect of temperature (20°C versus 26°C). A mixed-family approach was used in order to infer the genetic composition of larval populations and family assignment, limiting possible environmental bias and allowing the study of a relatively large number of families using a limited number of larval tanks. Our results show that three multiplexed highly polymorphic microsatellite markers are a powerful tool for family assignment and, consequently, for the study of bivalve larvae genetics. Culling, by selective sieving of the smallest larvae is an advantageous practice at a phenotypic scale as it reduced variance in larval size, variance of developmental rate and time to settlement. Culling of 50% of the larval population only led to 15% less spat, showing a positive phenotypic correlation between larval growth and settlement success. However, culling represents a substantial risk for diversity loss, because it increases the variance of reproductive success among parental oysters. The effective population sizes of early settling cohorts of settlement were lower than those of later ones. Our results show that the settlement of slow growing larvae significantly contributes to minimizing the variability of reproductive success and therefore to maximizing genetic diversity. These results corroborate the low estimations of variability of broodstocks sampled in several French commercial hatcheries, relative to natural populations. The genetic composition of the larval population and the resulting spat was significantly different between the two tested temperatures, revealing genotype x environment interaction for survival. Similarly, genotype x environment interaction was also observed for larval growth as a higher temperature exerted a positive influence on the expression of genetic variability for this trait. Consequently, we can conclude that a temperature of 26°C coupled with culling, to common practice in oyster hatcheries, is likely to amplify the selection pressure for fast growing larvae. To test for this hypothesis, we compared larval developmental traits in the progeny of a hatchery broodstock closed for 7 generations, with the progeny of wild oysters and the two possible hybrids. Our results show that selection of fast growing larvae can counteract presumed inbreeding depression, due to higher mean relatedness among hatchery broodstock than in the wild. Genetic effects of intensive rearing conditions at larval stage are significant and should be taken into account in hatchery practices, especially in terms of genetic diversity management.
Afin d'étudier les conséquences génétiques des pratiques de production en écloserie d'huître creuse, deux facteurs ont été examinés : l'élimination des petites larves et la température. Nos résultats montrent que l'assignation de parenté par marqueurs microsatellites est un outil performant pour les études génétiques en phase larvaire de familles élevées en mélange. Bien qu'avantageux d'un point de vue phénotypique, le tamisage sélectif représente un risque de perte de diversité. La fixation des larves à croissance lente permet en effet de minimiser la variabilité du succès reproducteur et de fait, de maximiser la variabilité génétique. Ces résultats corroborent les estimations de variabilité sur les stocks d'écloseries françaises où l'on constate une diversité allélique inférieure à celle de populations issues du milieu naturel. La température exerce également une influence sur la précocité de l'expression de la variabilité génétique pour la croissance larvaire. Ainsi une température élevée associée à une procédure de tamisage peut amplifier l'effet sélectif. Enfin, la sélection de larves à croissance rapide semble démontrée, s'opposant à la dépression de consanguinité présumée. Les conditions d'élevage peuvent donc avoir un effet génétique significatif qui devrait être pris en considération dans les pratiques d'écloserie, notamment dans la gestion de la diversité génétique.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2006/acte-1505.pdf
http://archimer.ifremer.fr/doc/00000/1505/

Résumé : Genetic consequences of production of Pacific oyster larval in hatchery: drift and selective pressures related to rearing practices. In order to study the genetic consequences of production of oyster larvae in hatcheries, two factors were examined: the effects of discarding the smallest larvae (i.e. culling) and temperature effects. A mixed-family approach was used in order to infer the genetic composition of the larval population. The results show that high polymorphic microsatellite-based family assignment is a powerful tool for the study of bivalve larvae genetics. Culling by selective sieving is an advantageous practice at a phenotypic scale, but also represents a substantial risk for diversity loss if parentage assignment is not introduced as a breeding practice. Settlement of slow growing larvae contributes to minimizing the variability of reproductive success and therefore to maximizing genetic diversity. These results corroborate the lower estimations of variability made on broodstocks from French commercial hatcheries relative to natural populations. Temperature exerts an influence on the expression of genetic variability for larval growth. A temperature of 26°C, coupled with culling could amplify the selective effect. Furthermore, selection of fast growing larvae has proven to counteract inbreeding depression at this stage. Genetic effects of intensive rearing conditions are significant and should be taken into account in hatchery practices, especially in terms of genetic diversity management.
Afin d'étudier les conséquences génétiques des pratiques de production de larves en écloserie d'huître creuse, deux facteurs ont été examinés : l'effet de l'élimination des plus petites larves et l'effet de la température. Une approche de familles élevées en mélange a été utilisée afin d'avoir accès à l'information génétique au stade larvaire. Les résultats obtenus montrent que l'assignation de parenté basée sur des marqueurs microsatellites hautement discriminants est un outil performant pour les études génétiques en phase larvaire. Bien qu'avantageuse d'un point de vue phénotypique, la pratique de tamisage sélectif représente un risque substantiel de perte de diversité si cette pratique n'est pas associée à une assignation de parentée par empreintes génétiques. La fixation des larves à croissance lente permet de minimiser la variabilité du succès reproducteur et de fait, de maximiser la variabilité génétique. Ces résultats corroborent les estimations de variabilité sur les stocks d'écloseries commerciales françaises où l'on constate une diversité allélique inférieure à celle de populations issues du milieu naturel. La température exerce également une influence sur la précocité de l'expression de la variabilité génétique pour la croissance larvaire. Ainsi une température élevée (26°C) associée à une procédure de tamisage peut amplifier l'effet sélectif. Enfin, la sélection de larves à croissance rapide semble démontrée, s'opposant à la dépression de consanguinité présumée en phase larvaire. Les conditions d'élevage peuvent donc avoir un effet génétique significatif qui devrait être pris en considération dans les pratiques d'écloserie, notamment dans la gestion de la diversité génétique.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2006/rapport-1459.pdf
http://archimer.ifremer.fr/doc/00000/1459/

Éditeur(s) : Pergamon-elsevier Science Ltd
Résumé : Metazooplankton abundance, biomass (<80 mu m, 200-500 mu m and >500 mu m) and community structure in the Ahe atoll were studied together with their relationships with environmental factors (temperature, salinity, wind) and trophic factors (phytoplankton, bacteria, heterotrophic nanoflagellates (HNF) and ciliates) during three periods in 2008-2009. Meroplankton, mainly bivalve and gastropod larvae, was dominant. Holoplankton was dominated by copepods, the main species being Oithona spp., Paracalanus parvus, Clausocalanus spp., Corycaeus spp., Acartia fossae and Undinula vulgaris. The results suggest a clear wind influence on the structure and horizontal distribution of the zooplankton communities. The metazooplankton appeared to be controlled mainly by food resources, suggesting a bottom-up control. The low nanophytoplankton biomass in contrast to the high abundance of picophytoplankton, HNF and nano-particle grazers (mainly Oithona spp., Paracalanus and bivalve larvae) highlighted the importance of the microbial loop in the food web. (C) 2012 Elsevier Ltd. All rights reserved.
Marine Pollution Bulletin (0025-326X) (Pergamon-elsevier Science Ltd), 2012 , Vol. 65 , N. 10-12 , P. 538-548

Droits : 2012 Elsevier Ltd. All rights reserved.
http://archimer.ifremer.fr/doc/00101/21207/19419.pdf
DOI:10.1016/j.marpolbul.2012.01.025
http://archimer.ifremer.fr/doc/00101/21207/

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
The embryonic development in Chione cancellata, from fertilization to straighthinge D-stage veliger larva, occurs in 24 h at 251C. Transmission (TEM) and scanning (SEM) electron microscopy show that morphogenetic processes result in a gastrula with two depressions, 4 h after fertilization (T014 h). Two hours later, one depression, located at the animal pole, develops into an open cave, the floor of which becomes the shell field, characterized by scarce microvilli, located below the lower face of the prototrochal pad. The invagination located at the vegetal pole is characterized by regular microvilli and constitutes the digestive tract. At T018 h, the late gastrula differentiates into a typical motile trochophore, possessing a well-developed prototroch delimiting anterior and posterior embryonic regions. At T019h, the shell field, located between the prototroch and the telotroch, appears as a saddle-shaped region with a wrinkled surface extending on both sides of the embryo, establishing bilateral symmetry. From TEM observations, the apical surface of the underlying cells possesses a modified glycocalyx featuring a dense pellicle overlapping the cells of the shell field. At T0112 h, the prototroch is displaced toward the anterior region by outgrowth of the shell material. TEM views show that the wrinkled surface observed with SEM corresponds to the periostracum, attached dorsally to the hinge and ventrally to the cells of the future mantle edge. At T0115 h, an alveolar matrix of proteinaceous material, which could contain calcite crystals, appears between the periostracum and the mantle. At T0124 h, prodissoconch I formation is completed and the D-stage larvae possess a calcified shell. At this stage of development, only one mantle fold is present, representing the outer fold. The periostracumsecreting cells are in contact with the marginal cells of the velum. This functional velum is composed of four bands of cilia. Our TEMobservations confirm previous descriptions of the velum ciliary bands obtained only from SEM analyses, such as described in other bivalves. The apical sense organ and the digestive tract are poorly developed. From observations at the ultrastructural level of shell differentiation in C. cancellata, we propose a new interpretation of shell differentiation, including hinge and ligament for bivalves as opposed to that described for gastropods.
ISSN: 1077-8306

hal-00755283
https://hal.univ-antilles.fr/hal-00755283
DOI : 10.1111/j.1744-7410.2006.00036.x

Éditeur(s) : HAL CCSD Station Biologique
Résumé : International audience
The embryonic development of Tivela mactroides, from fertilization to straight-hinge veliger D-stage larva occurs in 18 hours at 25°C. Scanning electronic observations show that morphogenetic processes result in a gastrula with two depressions 4 hours after fertilization (T0 + 4h). Two hours later, one depression, located at the animal pole, develops into an open cave, the floor of which becomes the shell field located below the lower face of the prototrochal pad. The invagination located at the vegetal pole features the blastopore. At T0 + 6h, the late gastrula has differentiated into a typical motile trochophore with a shell field synthetizing the organic part of the shell. At T0 + 8h, the shell field, located between the prototroch and the telotroch, appears as a saddle-shaped region with a wrinkled surface extending on both sides of the embryo, establishing bilateral symmetry. At T0 + 12h, the prototroch slides toward the anterior region by outgrowth of the shell material. At T0 + 18h, the prodissoconch I formation is completed and the D-stage larvae possess a calcified shell. At this stage of development, the functional velum is composed of four bands of cilia. Our observations seem to confirm the interpretation of shell differentiation proposed recently by Mouëza et al. (2006) for bivalves even if Transmission Electronic observations will be necessary to validate definitively such a model in the Meretricinae sub-family.
ISSN: 0007-9723

hal-00753022
https://hal.univ-antilles.fr/hal-00753022

Résumé : Le but de ce programme pathologie est de rechercher et d'étudier les agents pathogènes connus et inconnus chez les mollusques bivalves marins, plus particulièrement chez l'huître creuse, Crassostrea gigas, afin d'identifier d'éventuels agents causant des mortalités, de proposer des mesures prophylactiques et de définir des conditions zootechniques permettant aux professionnels de limiter l'impact des maladies sur leur cheptel. La présence de maladies, chez les mollusques bivalves marins dont l'impact socio économique peut être catastrophique pour les productions conchylicoles, et l'accroissement des échanges de produits d'aquaculture entre les pays de la CEE et avec les pays tiers nécessitent le renforcement des recherches dans les domaines de la pathologie. En particulier, du fait du mono élevage de l'huître japonaise, Crassostrea gigas, en France, il semble aujourd'hui indispensable d'effectuer des contrôles zoosanitaires réguliers chez cette espèce. Ces travaux ont d'autant plus d'importance qu'aujourd'hui, devant les pathologies rencontrées, les professionnels restent extrêmement démunis (traitements inapplicables ou inexistants). Dans ce cadre, la promptitude du diagnostic ainsi que l'acquisition de données épidémiologiques et biologiques concernant les agents pathogènes impliqués sont les meilleurs moyens d'éviter leur dissémination chez les mollusques bivalves marins. Les travaux entrepris dans ce cadre par l'URPIG, semblent d'autant plus indispensables qu'il n'existait pas jusqu'à ces dernières années de pathologies identifiées chez l'huître creuse, Crassostrea gigas, sur le littoral français. Cependant depuis 1991, sont apparus dans certaines écloseries françaises des épisodes de mortalité sur des larves de cette espèce. Il a été possible de détecter un virus apparenté à la famille des Herpesviridae par ses caractères morphologiques (NICOLAS et al., 1992) et de démontrer sa pathogénicité pour les larves (LE DEUFF et al., 1994). Par ailleurs, en 1993, 1994 et 1995, un virus de même type, a été observé en association à de fortes mortalités sur différents lots de naissain de la même espèce, originaires d'écloseries et de captage naturel. Par ailleurs, en 1993, suite à l'observation d'anomalies au niveau des branchies chez des huîtres creuses provenant de Charente Maritime et du bassin d'Arcachon associées à de médiocres performances de croissance, il a été entrepris des analyses de divers échantillons, en histologie classique et en microscopie électronique à transmission. Ces examens ont permis de révéler la présence au niveau des branchies, chez un nombre non négligeable d'animaux, d'un microorganisme de type chlamydien (RENAULT and COCHENNEC, 1995). Il a également été possible, chez cette même espèce, de détecter la présence d'un organisme de type rickettsien au niveau des branchies (RENAULT, COCHENNEC and CHOLLET, 1995).

Droits : 1995 Ifremer
http://archimer.ifremer.fr/doc/00043/15453/12828.pdf
http://archimer.ifremer.fr/doc/00043/15453/

Éditeur(s) : Pergamon-elsevier Science Ltd
Résumé : Bivalve larvae and hydrographic parameters were sampled over a range of spatio-temporal scales in a deep atoll lagoon. Bivalve larvae abundances were very high throughout the year: 18,550 m(-3) in average. Larvae were (i) concentrated at mid-depth with nocturnal ascent and diurnal descent, (ii) heterogeneously dispersed at the lagoon scale, (iii) subject to day-to-day variation in abundance and (iv) transferred between different parts of the lagoon providing evidence of intra-lagoonal connectivity. The primacy of physical factors was seen on large spatial scale with the diluting effect of water renewal and transfers by hydrodynamics. On smaller spatial scale, the primacy of biological processes was recognised, with larval swimming activity leading to dial vertical migration correlated with food concentration. Variations in larval abundance were driven by bivalve reproductive activity correlated with meteorological conditions (i.e. windy periods). Finally, relationship between bivalve larvae patterns and pearl oyster (Pinctada margaritifera) settlement structuring is discussed. (C) 2012 Elsevier Ltd. All rights reserved.
Marine Pollution Bulletin (0025-326X) (Pergamon-elsevier Science Ltd), 2012 , Vol. 65 , N. 10-12 , P. 453-462

Droits : 2012 Elsevier Ltd. All rights reserved.
http://archimer.ifremer.fr/doc/00101/21200/20690.pdf
DOI:10.1016/j.marpolbul.2011.12.028
http://archimer.ifremer.fr/doc/00101/21200/

Éditeur(s) : Springer
Résumé : The French Atlantic coast contains large highly productive intertidal mudflats that are colonised by juveniles of numerous flatfish species, including the common sole (Solea solea, L.). These ecosystems are also heavily exploited by the shellfish farming industry. Intensive bivalve culture is associated with substantial biodeposition (1-6 t-dw ha(-1) day(-1)), which directly or indirectly contributes to increase exopolysaccharide (EPS) concentrations at the interface between water column and seabed. EPS are long-chain molecules organised into colloids, which influence rheological properties of water, particularly viscosity. Increased water viscosity had consequences for ventilatory activity of juvenile flatfish, whereby the minimal pressure required to ventilate the medium increases directly with EPS concentration. Moreover, the critical EPS concentration ([EPS](crit)) at which water was no longer able to flow through the branchial basket ranged from almost nil to over 30 mg l(-1), depending on species and size. [EPS](crit) was lower in small individuals and individuals from species with high metabolic rates (turbot and plaice). These differences may depend upon gill and bucco-branchial cavity morphometrics. The ventilatory workload of sole increased with viscosity to a maximum at 2 mg EPS l(-1). Viscosity might, therefore, be a limiting factor for flatfish post larvae, which colonise the intertidal mudflats, depending upon their size and species. EPS concentrations in the field can reach 15 mg l(-1). A selective effect is conceivable but remains to be estimated in the field.
Marine Biology (0025-3162) (Springer), 2007-09 , Vol. 152 , N. 4 , P. 803-814

Droits : 2007 Springer
http://archimer.ifremer.fr/doc/2007/publication-2801.pdf
DOI:10.1007/s00227-007-0731-z
http://archimer.ifremer.fr/doc/00000/2801/

Éditeur(s) : Elsevier
Résumé : Vibrio tapetis is the marine bacterium responsible for the brown ring disease (BRD) affecting die manila clam, Ruditapes philippinarum. Identification of V tapelis has been previously performed using biochemical criteria and serological procedures. All of these methods are time consuming and ill-adapted to individual screening. This study describes an oligonucleotidic probe (Vt446) and two PCR primers, deduced from the 16S rDNA sequence, allowing a fast and specific V tapetis identification using dot blot hybridisation and species-specific primed PCR (SSP-PCR). The probe and primers have been tested on 60 strains, including referenced Vibrio sp., Gram negative and positive bacteria, marine bacteria samples and isolated clam bacteria. For all the 19 V tapetis strains, the results of PCR assays consistently corroborated those of the agglutination tests. The detection limit was estimated to be 10(2) CFU ml(-1). The SSP-PCR method has resulted in V tapetis detection in larvae, m diseased clams, and in asymptomatic broodstock clams that later developed BRD. In conclusion, the two SSP-PCR primers were useful for direct and fast identification of V tapetis strains isolated in clams, and are well suited for the screening of individual R. philippilnarum broodstock clams and larvae from the hatchery. (C) 2005 Elsevier B.V. All rights reserved.
Aquaculture (0044-8486) (Elsevier), 2006-03 , Vol. 253 , N. 1-4 , P. 25-38

Droits : 2005 Elsevier B.V. All rights reserved
http://archimer.ifremer.fr/doc/2006/publication-1692.pdf
DOI:10.1016/j.aquaculture.2005.03.047
http://archimer.ifremer.fr/doc/00000/1692/

Éditeur(s) : Inter-Research
Résumé : The marine environment is of special interest for studying hybridization between closely related taxa because of the high dispersal potential of planktonic larvae, such as those of most bivalve species. The oysters Crassostrea angulata and C. gigas are known to be very close genetically and entirely inter-fertile under controlled conditions. However, hybridization in the wild had not been investigated, mainly due to the lack of nuclear diagnostic markers. In the present paper, we first estimated genetic differentiation between these 2 closely related taxa using 8 microsatellite markers. Interestingly, 5 markers displayed significant differences of allele size between taxa. The subsequent sequencing of alleles of one of these microsatellites showed several mutational events, which suggested null alleles and homoplasy. The presence of 1 insertion/deletion event in its 5' flanking sequence enabled us to design a new bi-allelic ('C' and 'NC') nuclear PCR-restriction fragment length polymorphism (-RFLP) marker (CG44R). This, together with a mitochondrial DNA marker, was used to analyze populations of C. angulata and C. gigas. The CG44R allele frequencies were very different between C. angulata (f[C] = 0.91) and C. gigas (f[NC] = 0.92) populations. This analysis also provided evidence for hybridization between C. angulata and C. gigas in a wild Portuguese population where the 2 taxa are in contact due to recent transportation of C. gigas stocks for aquacultural production. Our results represent the first indication of hybridization between these 2 taxa in the natural environment, and contribute to knowledge of the evolutionary history of the Crassostrea genus.
Marine Ecology Progress Series (0171-8630) (Inter-Research), 2004 , Vol. 272 , P. 141-152

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2004/publication-3355.pdf
http://archimer.ifremer.fr/doc/00000/3355/

Éditeur(s) : Elsevier
Résumé : Deep-sea bivalves of the subfamily Bathymodiolinae (family Mytilidae) are very widespread and form dense beds in reduced environments such as hydrothermal vents and cold seeps. Bathymodiolus mussels recently discovered on African cold seeps strangely resemble Gulf of Mexico and Barbados seep species. This raises intriguing questions regarding their taxonomic relationships and their dispersal capabilities across the Atlantic equatorial belt. The morphological study of the shell and soft parts of mussels from either sites of the Atlantic shows that they form two distinct groups: the Bathymodiolus boomerang group (also including Bathymodiolus heckerae and a species from Africa), and the Bathymodiolus childressi group (also including Bathymodiolus mauritanicus and one species from Barbados). Phylogenetic relationships inferred from the nucleotide sequences of the ribosomal DNA internal transcribed spacer (ITS-2) and mitochondrial cytochrome c oxidase subunit I (COI) confirmed morphological analyses and the existence of two amphi-Atlantic complexes of species. Both ITS2 and COI phylogenies indicate almost no difference between the two eastern Atlantic seep mussels (Bathymodiolus sp. A and B. mauritanicus) and their western Atlantic counterparts (B. boomerang and Bathymodiolus sp. B; Barbados Prism cold seeps). In the B. boomerang complex, B. heckerae seems to differ from the Barbados and the African species, whereas these latter two are not distinguishable. In the B. childressi complex, relationships are less clear and do not support the description of new species from the Barbados. Past and present-day connections across the Atlantic are discussed in the light of both larval dispersal capabilities of the mussels and the equatorial Atlantic circulation to appreciate whether these species could represent true amphi-Atlantic species.
Deep Sea Research Part I: Oceanographic Research Papers (0967-0637) (Elsevier), 2007-11 , Vol. 54 , N. 11 , P. 1890-1911

Droits : 2007 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/2007/publication-4064.pdf
DOI:10.1016/j.dsr.2007.07.004
http://archimer.ifremer.fr/doc/00000/4064/

Résumé : Le but de ce programme pathologie est de rechercher et d'étudier les agents pathogènes connus et inconnus chez les mollusques bivalves marins, plus particulièrement chez l'huître creuse, Crassostrea gigas, afin d'identifier d'éventuels agents causant des mortalités, de proposer des mesures prophylactiques et de définir des conditions zootechniques permettant aux professionnels de limiter l'impact des maladies sur leur cheptel. La présence de maladies, chez les mollusques bivalves marins dont l'impact socio économique peut être catastrophique pour les productions conchylicoles, et l'accroissement des échanges de produits d'aquaculture entre les pays de la CEE et avec les pays tiers nécessitent le renforcement des recherches dans les domaines de la pathologie. En particulier, du fait du mono élevage de l'huître japonaise, Crassostrea gigas, en France, il est aujourd'hui indispensable d'effectuer des contrôles zoosanitaires réguliers chez cette espèce. Ces travaux ont d'autant plus d'importance qu'aujourd'hui, devant les pathologies rencontrées, les professionnels restent extrêmement démunis (traitements inapplicables ou inexistants). Dans ce cadre, la promptitude du diagnostic ainsi que l'acquisition de données épidémiologiques et biologiques concernant les agents pathogènes impliqués sont les meilleurs moyens d'éviter leur dissémination chez les mollusques bivalves marins. Les travaux entrepris dans ce cadre par le Laboratoire de Génétique, Aquaculture et Pathologie de La Tremblade, semblent d'autant plus indispensables qu'il n'existait pas jusqu'à ces dernières années de pathologies identifiées chez l'huître creuse, Crassostrea gigas, sur le littoral français. Cependant depuis 1991, sont apparus dans certaines écloseries françaises des épisodes de mortalité sur des larves de cette espèce. Il a été possible de détecter un virus apparenté à la famille des Herpesviridae par ses caractères morphologiques (NICOLAS et al., 1992) et de démontrer sa pathogénicité pour les larves (LE DEUFF et al., 1994). Par ailleurs, chaque année depuis 1993, un virus de même type, a été observé en association à de fortes mortalités sur certains lots de naissain de la même espèce, originaires d'écloseries et de captage naturel.

Droits : 1996 Ifremer
http://archimer.ifremer.fr/doc/00043/15452/12827.pdf
http://archimer.ifremer.fr/doc/00043/15452/