Production of triploid Kuruma shrimp, Marsupenaeus (Penaeus) japonicus (Bate) nauplii through inhibition of polar body I, or polar body I and II extrusion using 6-dimethylaminopurine

This study investigated the chromosome ploidy level of Marsupenaeus (Penaeus) japonicus (Bate) non-viable (unhatched) embryos and nauplii after exposure to 6-dimethylaminopurine (6-DMAP), timed to stop either polar body (PB) I, or PBI and II extrusion. Embryos from eight separate families or spawnings were exposed to 150 or 200 mu M 6-DMAP from 1- to 3-min post-spawning detection (psd) for a 4- to 5-min duration (timed to stop PBI extrusion). Separate aliquots of embryos from five of the same spawnings were also exposed to 200 mu M of 6-DMAP from 1- to 3-min psd for a 16-min duration (timed to stop both PBI and II extrusion). For one spawning, a third aliquot of embryos was exposed to 400 p M of 6-DMAP from 1- to 3-min psd for a 16-min duration (timed to stop both PBI and II extrusion). At 18-h psd, non-viable embryo and nauplii samples were taken separately for fluorescent activated cell sorting (FACS). FACS revealed that there were diploids and triploids among all treated non-viable embryos and nauplii. All control non-viable embryos and nauplii were diploid. Percentages of triploid induction for the 4- to 5-min and 16-min durations were not significantly different (P > 0.05). Additionally, no difference was found in the triploidy level of nonviable embryos compared to nauplii in these treatments. The percentage of triploid embryos and nauplii when exposed to 6-DMAP for a 4- to 5-min duration ranged from 29.57% to 99.23% (average 55.28 +/- 5.45%) and from 5.60% to 98.85% (average 46.70 +/- 7.20%), respectively. The percentage of triploid embryos and nauplii when exposed to 6-DMAP for a 16-min duration ranged from 11.71% to 98.96% (average 52.49 +/- 11.00%) and from 47.5% to 99.24% (average 79.38 +/- 5.24%), respectively. To our knowledge, this is the first documentation of successful PBI or PBI and II inhibition in shrimp. This study conclusively shows that treatment of M. japonicus embryos with 6-DMAP at 1- to 3-min pscl for either a 4- to 5-min duration (timed to stop PBl extrusion) or 16-min duration (timed to stop both PBI and II extrusion) results in viable triploid nauplii. (c) 2006 Elsevier B.V. All rights reserved.

Bilan 2015 du dispositif national de surveillance de la santé des mollusques marins

Depuis 1992, la surveillance de la santé des mollusques marins du littoral français est assurée par le réseau de Pathologie des Mollusques (Repamo). Ses activités s’inscrivent dans le cadre de la Directive Européenne 2006/88/CE. Depuis son évaluation par la plateforme nationale d’épidémiosurveillance en santé animale en 2012, l’objectif de surveillance est la détection précoce des infections dues à des organismes pathogènes exotiques et émergents affectant les mollusques marins sauvages et d’élevage. L’année 2015 est la première année de transition pour laquelle un début d’évolution des modalités de surveillance de la santé des mollusques marins animées par l’Ifremer a été amorcé. Un dispositif hybride de surveillance a été mis en place, s’appuyant sur l’existant et intégrant des débuts d’évolution. La surveillance événementielle a constitué l’activité principale du dispositif en 2015 et s’est appuyée sur des réseaux existants : (1) la surveillance des mortalités observées sur des animaux sentinelles déployés sur les sites ateliers des réseaux Ifremer RESCO 2 (12 sites) pour l’huître creuse Crassostrea gigas et MYTILOBS 2 (8 sites) pour la moule bleue Mytilus edulis. Pour l’huître creuse Crassostrea gigas, la mortalité cumulée moyenne était de 50,3% (écart-type 10,9%) pour le naissain standardisé Ifremer (NSI), de 11,0% (écart-type 9,1%) pour les huîtres de 18 mois et de 7,3% (écart-type 5,6%) pour les huîtres de 30 mois. Les mortalités ont été observées principalement entre le début du mois de mai et la mi-juillet. Lors de ces épisodes de mortalité, des prélèvements d’animaux ont été réalisés en vue d’analyses diagnostiques : 7 prélèvements pour le NSI, 2 pour les huîtres de 18 mois et 1 pour les huîtres de 30 mois. Aucun agent réglementé n’a été détecté dans les échantillons d’huîtres creuses prélevés et analysés. Le virus OsHV-1 a été détecté dans les 7 échantillons analysés de NSI, dans 2 échantillons analysés d’huîtres de 18 mois et dans 1 échantillon analysé d’huîtres de 30 mois. La bactérie Vibrio aestuarianus a été détectée dans 5 échantillons analysés de NSI, dans 1 échantillon d’huîtres de 18 mois et dans 1’échantillon d’huîtres de 30 mois. Pour la moule bleue Mytilus edulis, des mortalités cumulées variant de 9% sur le site du Vivier à 51% sur le site des filières du Pertuis Breton ont été estimées. Les mortalités ont été observées au printemps sur des moules âgées d’une année et en automne sur des moules plus jeunes. Lors de ces épisodes de mortalités, des prélèvements d’animaux ont été réalisés en vue d’analyses diagnostiques : 2 prélèvements pour les moules d’une année et 1 pour les jeunes moules. Ces prélèvements ont eu lieu dans le Pertuis Breton. Aucun agent réglementé n’a été détecté dans les échantillons de moules prélevés et analysés. Des bactéries du groupe Splendidus ont été détectées dans les 3 échantillons de moules analysés. (2) la surveillance s’appuyant sur les déclarations de mortalités de mollusques par les conchyliculteurs et pêcheurs à pied professionnels auprès des Directions départementales des territoires et de la mer (DDTM). Cette modalité s’applique aux huîtres creuses et aux moules bleues lorsqu’il n’existe pas de site atelier RESCO 2 ou MYTILOBS 2 dans la zone où des mortalités sont déclarées par les conchyliculteurs ou pêcheurs à pied. Le réseau REPAMO 2 a réalisé 22 interventions, dont 15 pour les moules Mytilus edulis, 4 pour les coques Cerastoderma edule, 2 pour les palourdes Ruditapes sp. et 1pour les coquilles saint Jacques Pecten maximus. La recherche d’agents infectieux dans ces espèces de mollusques prélevés lors de hausse de mortalité a permis de mettre en évidence les parasites réglementés Perkinsus olseni dans 1 lot de palourdes, et Marteilia refringens dans 4 lots de moules, ainsi que le virus OsHV-1 dans 1 lot de palourdes et 1 lot de coques, la bactérie Vibrio aestuarianus dans 3 lots de coques, et des bactéries du groupe Splendidus dans 3 lots de coques et dans 13 lots de moules. L’année 2015 a également permis la démonstration sur un site atelier d’un exercice de surveillance programmée, ciblée et fondée sur les risques d’introduction et d’installation d’un organisme pathogène exotique. Elle a concerné le parasite Mikrocytos mackini de l’huître creuse Crassostrea gigas, sur un site atelier de la Charente-Maritime, suivi par le réseau RESCO 2. Le parasite Mikrocytos mackini n’a pas été détecté. En revanche, le parasite Marteilia refringens a été détecté dans ¾ des prélèvements d’huîtres réalisés. Dans le cadre du soutien scientifique et technique de l’évolution de la surveillance événementielle, l’année 2015 a également permis de poursuivre la démarche relative aux développements méthodologiques en lien avec la surveillance événementielle des mortalités de mollusques marins. Une étude de faisabilité de la recherche prospective de regroupements spatio-temporels d’événements de mortalités d’huîtres creuses a été préparée en collaboration avec tous les acteurs de la santé des mollusques marins en Normandie. Un outil de collecte et d’analyse des données de signalements des mortalités, automatisé, simple d’utilisation et flexible, a été élaboré.

L'amélioration de la qualité des huîtres à échelle professionnelle: intégration de l'eau salée souterraine traitée comme milieu d'élevage

Les travaux menés au LCPL depuis plusieurs année sur l'engraissement contrôlé de l'huître creuse Crassostrea gigas ont permis, à l'échelle expérimentale de définir les différents paramètres d'élevage: Température: 14°C ration alimentaire: 2.109 cell/ind/j de la diatomée Skeletonema costatum indice AFNOR initial sans incidence débit d'eau: 3 lIind/h eau de mer ou eau salée souterraine Dans ces conditions et après 30 jours d'engraissement, le poids total des huîtres a progressé de 10%, le poids sec de 230%, l'indice AFNOR de 80% et la concentration en glycogène de 450%. Le travail réalisé ici a pour but de transposer de l'échelle expérimentale à une échelle significative pour les professionnels, la technique d'engraissement contrôlé en utilisant l'eau salée souterraine pour la production de phytoplancton d'une part et comme vecteur de régulation thermique d'autre part. En effet, un des intérêts de l'utilisation de l'eau salée souterraine réside dans sa température constante de 14°C permettant un engraissement des huîtres en toute saison sans risque de déclenchement de la gamétogenèse. Afin de vérifier cette possibilité, l'étude a été réalisée au cours de deux saisons: printemps et automne. Pour l'eassai automnal, trois bassins de 8 m2 ont été utilisés, alimentés respectivement en eau de mer naturelle, eau de mer régulé en température par échange thermique avec de l'eau salée souterraine et eau salée souterraine traitée. Dans chaque bassin, 330 kg d'huîtres ont été disposées en 7 ruches de 4 clayettes. Pour l'essai printanier, seuls deux bassins ont été utilisés, alimentés en eau de mer thermorégulée à deux débits différents: 0.61/h et 3 lIh. L'étude a été conduite pour chaque saison sur une période de 35 jours. En automne 2000, l'indice de qualité de chair de 8.2 initialement, a atteint 12.5 en eau salée souterraine, 13.8 en eau de mer thermorégulée et 14.2 en eau de mer naturelle. Le poids de chair sèche passe de 0.9 g initialement à respectivement 2.32g, 2.39g et 2.37g. Les dosages de Pb, Cd, Hg, Mn et As ne montent pas d'évolution entre le début et la fin de l'élevage, par contre le Fe augmente sensiblement dans les huîtres sur eau salée souterraine traitée. En fin d'élevage, une période de stockage de 4 semaines des huîtres sans nourriture, n'a pas montré de perte de qualité. Au printemps 2001, un deuxième essai sera conduit pour vérifier les résultats obtenus à l'automne 2000, confirmer le choix du milieu d'élevage et proposer les éléments permettant d'approcher les coûts de production.

Aeromonas spp. isolated from oysters (Crassostrea rhizophorea) from a natural oyster bed, Ceará, Brazil

Between April and October 2002, thirty fortnightly collections of oysters (Crassostrea rhizophorea) from a natural oyster bed at the Cocó River estuary in the Sabiaguaba region (Fortaleza, Ceará, Brazil) were carried out, aiming to isolate Aeromonas spp. strains. Oyster samples were submitted to the direct plating (DP) and the presence/absence (P/A) methods. Aeromonas were identified in 15 (50%) samples analyzed by the DP method and in 13 (43%) analyzed by the P/A method. A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii and Aeromonas sp. were isolated. The predominant species was A. veronii (both biovars), which was identified in 13 (43%) samples, followed by A. media in 11 (37%) and A. caviae in seven (23%). From the 59 strains identified, 28 (48%) presented resistance to at least one of the eight antibiotics tested.

Integrated treatment of shrimp effluent by sedimentation, oyster filtration and macroalgal absorption: a laboratory scale study

Effluent water from shrimp ponds typically contains elevated concentrations of dissolved nutrients and suspended particulates compared to influent water. Attempts to improve effluent water quality using filter feeding bivalves and macroalgae to reduce nutrients have previously been hampered by the high concentration of clay particles typically found in untreated pond effluent. These particles inhibit feeding in bivalves and reduce photosynthesis in macroalgae by increasing effluent turbidity. In a small-scale laboratory study, the effectiveness of a three-stage effluent treatment system was investigated. In the first stage, reduction in particle concentration occurred through natural sedimentation. In the second stage, filtration by the Sydney rock oyster, Saccostrea commercialis (Iredale and Roughley), further reduced the concentration of suspended particulates, including inorganic particles, phytoplankton, bacteria, and their associated nutrients. In the final stage, the macroalga, Gracilaria edulis (Gmelin) Silva, absorbed dissolved nutrients. Pond effluent was collected from a commercial shrimp farm, taken to an indoor culture facility and was left to settle for 24 h. Subsamples of water were then transferred into laboratory tanks stocked with oysters and maintained for 24 h, and then transferred to tanks containing macroalgae for another 24 h. Total suspended solid (TSS), chlorophyll a, total nitrogen (N), total phosphorus (P), NH4+, NO3-, and PO43-, and bacterial numbers were compared before and after each treatment at: 0 h (initial); 24 h (after sedimentation); 48 h (after oyster filtration); 72 h (after macroalgal absorption). The combined effect of the sequential treatments resulted in significant reductions in the concentrations of all parameters measured. High rates of nutrient regeneration were observed in the control tanks, which did not contain oysters or macroalgae. Conversely, significant reductions in nutrients and suspended particulates after sedimentation and biological treatment were observed. Overall, improvements in water quality (final percentage of the initial concentration) were as follows: TSS (12%); total N (28%); total P (14%); NH4+ (76%); NO3- (30%); PO43-(35%); bacteria (30%); and chlorophyll a (0.7%). Despite the probability of considerable differences in sedimentation, filtration and nutrient uptake rates when scaled to farm size, these results demonstrate that integrated treatment has the potential to significantly improve water quality of shrimp farm effluent. (C) 2001 Elsevier Science B.V. All rights reserved.

Coliform density in oyster culture waters and its relationship with environmental factors

The objective of this work was to evaluate the total and thermotolerant coliform densities in the oyster culture water of Cananeia, SP, Brazil, correlating these densities with environmental variables and tidal variations. Superficial water samples were collected in two tide conditions (spring and neap) from three areas of Cananéia municipality (Mandira, Itapitangui and Cooperostra). The three studied areas showed good conditions for the culture regarding coliform densities. The two tidal conditions differed significantly as to total coliform concentration; however, the same procedure was not performed for thermotolerant coliforms. No correlation was observed between water temperature, pH, and concentrations of total and thermotolerant coliforms. Coliform density was positively correlated with rainfall and negatively correlated with salinity. Spring and neap tides differed significantly as to coliform number. Simple diagnosis of environmental conditions of the crop fields is insufficient to assess water quality of shellfish cultivation. A continuous monitoring program of planted areas is necessary both for the assessment of water quality potential for marine culture and for ensuring safe consumption of seafood, besides constituting an important tool to understand the relationships between contamination and the involved environmental variables.

Crassostrea patagonica (d'Orbigny, 1842) shell concentrations from the late Miocene Rio Negro province, NE Patagonia, Argentina

The Río Negro Formation (late Miocene-early Pliocene) mainly consists of continental deposits, but it contains a middle member of marine origin. It represents a transgressive-regressive sequence that can be seen at several outcrops along the N Patagonian coast. The taphonomical approach to the El Espigón marine deposits permits the identification of four main layers containing different kinds of skeletal accumulation, which mainly consist of oyster shells [Crassostrea patagonica (D'Orbigny, 1842)]. These concentrations display three different morphologies (pouches, pavements and bouquets) with a different taphonomic signature. These deposits were formed in shallow marine environments influenced by wave activity that produced valve concentrations of different entities. They contain several shell beds that represent event, composite, hiatal to lag skeletal concentrations. Traces of bioturbation in the sediment (Thalassinoides, Teichichnus) and bioerosion on the shells (Entobia, Gastrochaeonolites, Caulostrepsis), and encrusters (cirripeds, bryozoans), are also abundant in the outcrop and consititue common components of these Miocene materials. Layers 1 and 2 of the sequence were deposited in shoreface/foreshore environments at the beginning of a highstand systems tract, while layers 3 and 4 were deposited at the end, or at the beginning of a forced regression, in foreshore environments. A final erosional episode cut the top of the layer 4, which truncated the abundant bioturbaation developed there.

Studies on the Back Water Oyster, grassostrea madra— sensis (Preston) of Cochin Harbour

A detailed study of the hydrography of the Cochin Backwaters, the habitat off crassostrea madrasensis has been carried out. Data pertaining to air temperature, water temperature, salinity, dissolved oxygen, turbidity and rainfall have been collected and presented. The temperature fluctuation was in the range of 5°C only and that of salinity between 1.1%o and 32.9%o. Fairly steady salinity has been recorded during the pre-monsoon period (February to May) and drastic declension during the monsoon period (June-September).Dissolved oxygen varied between 2.5 ml/l and 6.5 ml/l. Turbidity was highest in June (27.9 p.p.m.) and minimum (10.2 p.p.m.) in February. A detailed study on marine biofouling in the Cochin Backwaters has been made with special reference to primary film, settlement and growth of the fouling organisms such as hydroids, bryozoans, tube-dwelling polychaetes, barnacles and modiolus

Coliform density in oyster culture waters and its relationship with environmental factors

The objective of this work was to evaluate the total and thermotolerant coliform densities in the oyster culture water of Cananeia, SP, Brazil, correlating these densities with environmental variables and tidal variations. Superficial water samples were collected in two tide conditions (spring and neap) from three areas of Cananéia municipality (Mandira, Itapitangui and Cooperostra). The three studied areas showed good conditions for the culture regarding coliform densities. The two tidal conditions differed significantly as to total coliform concentration; however, the same procedure was not performed for thermotolerant coliforms. No correlation was observed between water temperature, pH, and concentrations of total and thermotolerant coliforms. Coliform density was positively correlated with rainfall and negatively correlated with salinity. Spring and neap tides differed significantly as to coliform number. Simple diagnosis of environmental conditions of the crop fields is insufficient to assess water quality of shellfish cultivation. A continuous monitoring program of planted areas is necessary both for the assessment of water quality potential for marine culture and for ensuring safe consumption of seafood, besides constituting an important tool to understand the relationships between contamination and the involved environmental variables.

Seawater carbonate chemistry and biological processes of oysters Crassostrea virginica during experiments, 2010

Estuarine organisms are exposed to periodic strong fluctuations in seawater pH driven by biological carbon dioxide (CO2) production, which may in the future be further exacerbated by the ocean acidification associated with the global rise in CO2. Calcium carbonate-producing marine species such as mollusks are expected to be vulnerable to acidification of estuarine waters, since elevated CO2 concentration and lower pH lead to a decrease in the degree of saturation of water with respect to calcium carbonate, potentially affecting biomineralization. Our study demonstrates that the increase in CO2 partial pressure (pCO2) in seawater and associated decrease in pH within the environmentally relevant range for estuaries have negative effects on physiology, rates of shell deposition and mechanical properties of the shells of eastern oysters Crassostrea virginica (Gmelin). High CO2 levels (pH ~7.5, pCO2 ~3500 µatm) caused significant increases in juvenile mortality rates and inhibited both shell and soft-body growth compared to the control conditions (pH ~8.2, pCO2 ~380 µatm). Furthermore, elevated CO2 concentrations resulted in higher standard metabolic rates in oyster juveniles, likely due to the higher energy cost of homeostasis. The high CO2 conditions also led to changes in the ultrastructure and mechanical properties of shells, including increased thickness of the calcite laths within the hypostracum and reduced hardness and fracture toughness of the shells, indicating that elevated CO2 levels have negative effects on the biomineralization process. These data strongly suggest that the rise in CO2 can impact physiology and biomineralization in marine calcifiers such as eastern oysters, threatening their survival and potentially leading to profound ecological and economic impacts in estuarine ecosystems.

Mark-recapture may reveal more about ecology than about population trends: Demography of a threatened ghost bat (Macroderma gigas) population

Since European settlement in Australia, the geographical range of ghost bats (Macroderma gigas) has contracted northwards. Ghost bats are thought to occur in disjunct populations with little interpopulation migration, raising concerns over the current status and future viability of the southernmost colony, which has also been threatened by mining activity. To address these concerns, demographic parameters of the southernmost colony were estimated from a mark-recapture study conducted during 1975-1981. Female bats gave birth to a single young in late spring, but only 40% (22-70%, 95% CI) of females bred in their second year, increasing to 93% (87-97%, 95% CI) for females greater than or equal to 2 years old. Sixty-five percent of juveniles caught were female. Annual adult survival ranged between 0.57-0.77 for females and 0.43-0.66 for males, and was lowest over winter-spring and greatest in autumn-winter. Juvenile survival for the first year ranged between 0.35-0.46 for females and 0.29-0.42 for males. Adult survival varied among seasons, was negatively associated with rainfall, but was not associated with temperature beyond being lower in late winter. Poor survival may result from the inferior daytime roosts that bats must use if water seepage forces them to leave their normal roosts. Although these age-specific rates of fecundity and survival suggested a declining population, mark-recapture estimates of the population trend indicated stability over the study period. Counts at daytime roosts also suggested a population decline, but were considered unreliable because of an increasing tendency of bats to avoid detection. It is therefore likely that some assumptions in estimating survival were violated. These results provide a caution against the uncritical use of population projections derived from mark-recapture estimates of demographic parameters, and the use of untested indices as the basis for conservation decisions.

Structural stability of adenylate kinase from the sulfate-reducing bacteria Desulfovibrio gigas

Biophysical Chemistry 110 (2004) 83–92

Zinc-substituted desulfovibrio gigas desulforedoxins

Protein Science (2002), 11:2464–2470

Direct electrochemistry of the Desulfovibrio gigas aldehyde oxidoreductase

Eur. J. Biochem. 271, 1329–1338 (2004)