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.

Intake rates and the functional response in shorebirds (Charadriiformes) eating macro-invertebrates

As field determinations take much effort, it would be useful to be able to predict easily the coefficients describing the functional response of free-living predators, the function relating food intake rate to the abundance of food organisms in the environment. As a means easily to parameterise an individual-based model of shorebird Charadriiformes populations, we attempted this for shorebirds eating macro-invertebrates. Intake rate is measured as the ash-free dry mass (AFDM) per second of active foraging; i.e. excluding time spent on digestive pauses and other activities, such as preening. The present and previous studies show that the general shape of the functional response in shorebirds eating approximately the same size of prey across the full range of prey density is a decelerating rise to a plateau, thus approximating the Holling type 11 ('disc equation') formulation. But field studies confirmed that the asymptote was not set by handling time, as assumed by the disc equation, because only about half the foraging time was spent in successfully or unsuccessfully attacking and handling prey, the rest being devoted to searching. A review of 30 functional responses showed that intake rate in free-living shorebirds varied independently of prey density over a wide range, with the asymptote being reached at very low prey densities (< 150/m(-2)). Accordingly, most of the many studies of shorebird intake rate have probably been conducted at or near the asymptote of the functional response, suggesting that equations that predict intake rate should also predict the asymptote. A multivariate analysis of 468 'spot' estimates of intake rates from 26 shorebirds identified ten variables, representing prey and shorebird characteristics, that accounted for 81 % of the variance in logarithm-transformed intake rate. But four-variables accounted for almost as much (77.3 %), these being bird size, prey size, whether the bird was an oystercatcher Haematopus ostralegus eating mussels Mytilus edulis, or breeding. The four variable equation under-predicted, on average, the observed 30 estimates of the asymptote by 11.6%, but this discrepancy was reduced to 0.2% when two suspect estimates from one early study in the 1960s were removed. The equation therefore predicted the observed asymptote very successfully in 93 % of cases. We conclude that the asymptote can be reliably predicted from just four easily measured variables. Indeed, if the birds are not breeding and are not oystercatchers eating mussels, reliable predictions can be obtained using just two variables, bird and prey sizes. A multivariate analysis of 23 estimates of the half-asymptote constant suggested they were smaller when prey were small but greater when the birds were large, especially in oystercatchers. The resulting equation could be used to predict the half-asymptote constant, but its predictive power has yet to be tested. As well as predicting the asymptote of the functional response, the equations will enable research workers engaged in many areas of shorebird ecology and behaviour to estimate intake rate without the need for conventional time-consuming field studies, including species for which it has not yet proved possible to measure intake rate in the field.

Data compilation on the biological response to ocean acidification: environmental and experimental context of data sets and related literature

The exponential growth of studies on the biological response to ocean acidification over the last few decades has generated a large amount of data. To facilitate data comparison, a data compilation hosted at the data publisher PANGAEA was initiated in 2008 and is updated on a regular basis (doi:10.1594/PANGAEA.149999). By January 2015, a total of 581 data sets (over 4 000 000 data points) from 539 papers had been archived. Here we present the developments of this data compilation five years since its first description by Nisumaa et al. (2010). Most of study sites from which data archived are still in the Northern Hemisphere and the number of archived data from studies from the Southern Hemisphere and polar oceans are still relatively low. Data from 60 studies that investigated the response of a mix of organisms or natural communities were all added after 2010, indicating a welcomed shift from the study of individual organisms to communities and ecosystems. The initial imbalance of considerably more data archived on calcification and primary production than on other processes has improved. There is also a clear tendency towards more data archived from multifactorial studies after 2010. For easier and more effective access to ocean acidification data, the ocean acidification community is strongly encouraged to contribute to the data archiving effort, and help develop standard vocabularies describing the variables and define best practices for archiving ocean acidification data.

Immunostimulatory effects of different aspects of aquaculture on the host response in the edible sea urchin, paracentrotus lividus

Aquaculture is a fast-growing industry contributing to global food security and sustainable aquaculture, which may reduce pressures on capture fisheries. The overall objective of this thesis was to look at the immunostimulatory effects of different aspects of aquaculture on the host response of the edible sea urchin, Paracentrotus lividus, which are a prized delicacy (roe) in many Asian and Mediterranean countries. In Chapter 1, the importance of understanding the biology, ecology, and physiology of P. lividus, as well as the current status in the culture of this organism for mass production and introducing the thesis objectives for following chapters is discussed. As the research commenced, the difficulties of identifying individuals for repeat sampling became clear; therefore, Chapter 2 was a tagging experiment that indicated PIT tagging was a successful way of identifying individual sea urchins over time with a high tag retention rate. However, it was also found that repeat sampling via syringe to measure host response of an individual caused stress which masked results and thus animals would be sampled and sacrificed going forward. Additionally, from personal observations and discussion with peers, it was suggested to look at the effect that diet has on sea urchin immune function and the parameters I measured which led to Chapter 3. In this chapter, both Laminaria digitata and Mytilus edulis were shown to influence measured immune parameters of differential cell counts, nitric oxide production, and lysozyme activity. Therefore, trials commencing after Trial 5 in Chapter 4, were modified to include starvation in order to remove any effect of diet. Another important aspect of culturing any organism is the study of their immune function and its response to several immunostimulatory agents (Chapter 4). Zymosan A was shown to be an effective immunostimulatory agent in P. lividus. Further work on handled/stored animals (Chapter 5) showed Zymosan A reduced the measured levels of some immune parameters measured relative to the control, which may reduce the amount of stress in the animals. In Chapter 6, animals were infected with Vibrio anguillarum and, although V. anguillarum, impacted immune parameters of P. lividus, it did not cause mortality as predicted. Lastly, throughout this thesis work, it was noted that the immune parameters measured produced different values at different times of the year (Chapter 7); therefore, using collated baseline (control) data, results were compiled to observe seasonal effects. It was determined that both seasonality and sourcing sites influenced immune parameter measurements taken at different times throughout the year. In conclusion, this thesis work fits into the framework of development of aquaculture practices that affect immune function of the host and future research focusing on the edible sea urchin, P. lividus.

Impacts of seawater acidification on mantle gene expression patterns of the Baltic Sea blue mussel: implications for shell formation and energy metabolism, link to supplementary material

Marine organisms have to cope with increasing CO2 partial pressures and decreasing pH in the oceans. We elucidated the impacts of an 8-week acclimation period to four seawater pCO2 treatments (39, 113, 243 and 405 Pa/385, 1,120, 2,400 and 4,000 µatm) on mantle gene expression patterns in the blue mussel Mytilus edulis from the Baltic Sea. Based on the M. edulis mantle tissue transcriptome, the expression of several genes involved in metabolism, calcification and stress responses was assessed in the outer (marginal and pallial zone) and the inner mantle tissues (central zone) using quantitative real-time PCR. The expression of genes involved in energy and protein metabolism (F-ATPase, hexokinase and elongation factor alpha) was strongly affected by acclimation to moderately elevated CO2 partial pressures. Expression of a chitinase, potentially important for the calcification process, was strongly depressed (maximum ninefold), correlating with a linear decrease in shell growth observed in the experimental animals. Interestingly, shell matrix protein candidate genes were less affected by CO2 in both tissues. A compensatory process toward enhanced shell protection is indicated by a massive increase in the expression of tyrosinase, a gene involved in periostracum formation (maximum 220-fold). Using correlation matrices and a force-directed layout network graph, we were able to uncover possible underlying regulatory networks and the connections between different pathways, thereby providing a molecular basis of observed changes in animal physiology in response to ocean acidification.

(Table 3, Page 417) Table 3. The concentrations of Zn, Cu, Pb, Cd, Ni, Co, Ag, Mn and Fe in a soluble in 1M HCl fraction of ferromanganese nodules of Slupsk Furrow in the southern Baltic sea

The concentration of Zn, Cu, Pb, Cd, Ni, Co, Ag, Mn, Fe, Ca, Mg, K and Na in molluscs Macoma balthica, Mya arenaria, Cardium glaucum, Mytilus edulis and Astarte borealis from the southern Baltic was determined. The surface sediments and ferromanganese concretions associated with the molluscs were also analysed for concentration of these metals. Species- and region-dependent differences in the metal levels of the organisms were observed. The properties of molluscs analysed which have a tendency toward elevated biological tolerance of selected trace metals were specified. The interelement relationship between metal concentrations in the soft tissue and the shell was estimated and was discussed.

The influence of mussel-modified habitat on Fucus serratus L. a rocky intertidal canopy-forming macroalga

The influence of habitat modification by Mytilus edulis L. on the settlement and development of Fucus serratus populations was investigated on rocky shores of the Isle of Anglesey, North Wales. Settlement of fucoids was higher inside mussel habitat than outside on one of two shores studied. The effect of microhabitat on survival of fucoid germlings was examined by transplanting the germlings into and outside mussel habitats, each with and without the exclusion of grazers. Observation showed that periwinkles and top shells were abundant in mussel habitat, while limpets dominated bare rock. Exclusion of grazers greatly enhanced the survival of fucoid germlings in both habitats, indicating that while mussel habitat supports a different grazer assemblage to bare rock, both assemblages are important in limiting fucoid recruitment. The risk of dislodgement was assessed and compared between fucoids growing on mussel shells and bare rock. In situ pull-tests showed that less force was required to detach large fertile thalli growing on mussel shells than those growing on the rock. Adhesion was generally broken between the mussel and the rock rather than between the holdfast and the mussel. These observations indicate that mussels provide an unstable substrate for mature fucoids. Overall results suggest that a negative effect of mussel-modified habitat on fucoids is profound in adults; but the effect is context-dependent in juveniles and can be positive at settlement. Results from a survey on population structure of fucoids across two shores showed that there were greater numbers of large fertile fucoids growing directly attached to rock than on mussel shells, while there was no difference for juvenile fucoids confirming the experimental results. Moreover thalli larger than 60 cm were found only on the rock but not on shells. This finding suggests that a mussel dominated habitat may have a significant impact on reproductive output in fucoid populations.

The influence of mussel-modified habitat on Fucus serratus L. a rocky intertidal canopy-forming macroalga

The influence of habitat modification by Mytilus edulis L. on the settlement and development of Fucus serratus populations was investigated on rocky shores of the Isle of Anglesey, North Wales. Settlement of fucoids was higher inside mussel habitat than outside on one of two shores studied. The effect of microhabitat on survival of fucoid germlings was examined by transplanting the germlings into and outside mussel habitats, each with and without the exclusion of grazers. Observation showed that periwinkles and top shells were abundant in mussel habitat, while limpets dominated bare rock. Exclusion of grazers greatly enhanced the survival of fucoid germlings in both habitats, indicating that while mussel habitat supports a different grazer assemblage to bare rock, both assemblages are important in limiting fucoid recruitment. The risk of dislodgement was assessed and compared between fucoids growing on mussel shells and bare rock. In situ pull-tests showed that less force was required to detach large fertile thalli growing on mussel shells than those growing on the rock. Adhesion was generally broken between the mussel and the rock rather than between the holdfast and the mussel. These observations indicate that mussels provide an unstable substrate for mature fucoids. Overall results suggest that a negative effect of mussel-modified habitat on fucoids is profound in adults; but the effect is context-dependent in juveniles and can be positive at settlement. Results from a survey on population structure of fucoids across two shores showed that there were greater numbers of large fertile fucoids growing directly attached to rock than on mussel shells, while there was no difference for juvenile fucoids confirming the experimental results. Moreover thalli larger than 60 cm were found only on the rock but not on shells. This finding suggests that a mussel dominated habitat may have a significant impact on reproductive output in fucoid populations.

Benthic assemblages associated with native and non-native oysters are similar

Invasive species can impact native species and alter assemblage structure, which affects associated ecosystem functioning. The pervasive Pacific oyster, Crassostrea gigas, has been shown to affect the diversity and composition of many host ecosystems. We tested for effects of the presence of the invasive C. gigas on native assemblages by comparing them directly to assemblages associated with the declining native European oyster, Ostrea edulis. The presence of both oyster species was manipulated in intertidal and subtidal habitats and reefs were constructed at horizontal and vertical orientation to the substratum. After 12 months, species diversity and benthic assemblage structure between assemblages with C. gigas and O. edulis were similar, but differed between habitats and orientation, suggesting that both oyster species were functionally similar in terms of biodiversity facilitation. These findings support evidence, that non-native species could play an important role in maintaining biodiversity in systems with declining populations of native species.

Apport d'un programme de génétique à une filière de production aquacole : l'exemple de l'ostréiculture

Two oyster species are currently present along the French coasts : the indigenous European flat oyster (Ostrea edulis), and the Pacific cupped oyster (Crassostrea gigas), that has been introduced from Japan since the beginning of the 70ies. The flat oyster successively suffered from two protozoan diseases during the 60ies and its production decreased from 20 000 tons/year by that time to 1 500 tons/year nowadays. Consequently, the oyster production is principally (99%) based upon the Pacific oyster species with approximately 150 000 tons/year among which 90% are grown from the natural spat. However, the hatchery production of this species is developing and was estimated to 400 to 800 millions spat in 2002. Moreover, strengthened relationships between IFREMER and the 5 commercial hatcheries, that all joined the SYSAAF (Union of the French poultry, shellfish and fish farming selectors), allow to plan for new genetic breeding programs. At the end of the 80ies, IFREMER initiated a genetic breeding program for the resistance of the European flat oyster to the bonamiosis, and obtained strains more tolerant to this disease. After two generations of massal selection, molecular markers had identified a reduced genetic basis in this program. It was then reoriented to an intra-familial selection. However, we were confronted to a zootechnic problem to manage such a scheme and we compromised by an intra-cohorts of families selection scheme managed using molecular markers. The program has now reached the transfer level with experimentation at a professional scale. Concerning the Pacific cupped oyster, and in parallel with the obtaining and the study of polyploids, performance of different Asian cupped oyster strains were compared to the one introduced in France thirty years ago and currently suffering from summer mortalities. The local strain exhibited better performance, certainly based upon a good local adaptation. In other respects, although early growth is a relevant criteria for selection for growth to commercial stage, it is not to be privileged in the context of an oyster producing region with a limited food availability. Contrary, the spat summer mortality became a priority for numerous teams (genetic, physiology, pathology, ecology,...) joined in the MOREST program. The first results showed important survival differences between fullsib and halsib families. They indicate a genetic determinism to this character "survival" and promote for its selection.

Can survival of European flat oysters following experimental infection with Bonamia ostreae be predicted using QTLs?

The present study identifies quantitative trait loci (QTLs) in response to an experimental infection with the parasite responsible for bonamiosis, Bonamia ostreae, in two segregating families of the European flat oyster, Ostrea edulis. We first constructed a genetic-linkage map for each studied family and improved the existing genetic-linkage map for the European flat oyster with a set of SNP markers. This latter map now combines the best accuracy and the best estimate of the genome coverage available for an oyster species. Secondly, by comparing the QTLs detected in this study with those previously published for O. edulis in similar experimental conditions, we identified several potential QTLs that were identical between the different families, and also new specific QTLs. We also detected, within the confidence interval of several QTL regions, some previously predicted candidate genes differentially expressed during an infection with B. ostreae, providing new candidate genome regions which should now be studied more specifically.

Climate Change Influences Carrying Capacity in a Coastal Embayment Dedicated to Shellfish Aquaculture

A spatially explicit coupled hydrodynamic-biogeochemical model was developed to study a coastal ecosystem under the combined effects of mussel aquaculture, nutrient loading and climate change. The model was applied to St Peter's Bay (SPB), Prince Edward Island, Eastern Canada. Approximately 40 % of the SPB area is dedicated to mussel (Mytilus edulis) longline culture. Results indicate that the two main food sources for mussels, phytoplankton and organic detritus, are most depleted in the central part of the embayment. Results also suggest that the system is near its ultimate capacity, a state where the energy cycle is restricted to nitrogen-phytoplankton-detritus-mussels with few resources left to be transferred to higher trophic levels. Annually, mussel meat harvesting extracts nitrogen (N) resources equivalent to 42 % of river inputs or 46.5 % of the net phytoplankton primary production. Under such extractive pressure, the phytoplankton biomass is being curtailed to 1980's levels when aquaculture was not yet developed and N loading was half the present level. Current mussel stocks also decrease bay-scale sedimentation rates by 14 %. Finally, a climate change scenario (year 2050) predicted a 30 % increase in mussel production, largely driven by more efficient utilization of the phytoplankton spring bloom. However, the predicted elevated summer temperatures (> 25 A degrees C) may also have deleterious physiological effects on mussels and possibly increase summer mortality levels. In conclusion, cultivated bivalves may play an important role in remediating the negative impacts of land-derived nutrient loading. Climate change may lead to increases in production and ecological carrying capacity as long as the cultivated species can tolerate warmer summer conditions.

Site n°5. The Pertuis Breton (Bay of Biscay) France

1.This report presents the results of a field study conducted in the ECASA test site nOS in the Pertuis Breton, France. The site is located on the Atlantic West coasts. It is open to the bay of Biscay, but is slightly protected against westerly winds. The bay has been exploited by intertidal mussels culture for centuries. 2. Within the bay, mussels (Mytilus edulis) are cultivated either by the traditional pole technique, around the bay or on longlines in the centre of the bay. The area occupied by these longline s represents 250 ha, and the resulting annual production is 1 000 tonnes of mussels. The average depth at mid tide is of 13.8 m. The sediment is sandy, with a small fraction of mud. 3. The site is subject to several regular monitoring through the local implementation of national networks aiming at protecting the environment and marine resources, on pollutants (RNO), microbiological quality of the waters (REMI), phytoplanktonic toxic species (REPHY) and growth and mortality of molluscs (REMORA). Benthic macrofauna was studied in 1976. 4. Five sampling sations were chosen along a line, starting under the longlines, and at distances of 50, 100, 200, and 400 metres from the area cultivated. A reference station was chosen in a different direction at 2300 metres of the cultivated area. Sampling methods are described in the text. _Sediments were sampled for different analyses: grain size, content in organic matter, total organic carbon and nitrogen, and phytic pigments (chlorophyll a and phaeopigments). Redox were measured in cores. The macrofauna living into the sediment was also sampled. The water column was sampled for physical (temperature, transparency) and chemical parametres, including oxygen content, salinity, organic matter, dissolved nitrogen forms, phosphates and silicates. Results from benthic macrofauna surveys indicate that there were no significant differences between the different stations and the reference station, all being classified as slightly disturbed. The bay is submitted to freshwater runoffs from two adjacent rivers. 7. The sediment is slightly modified by the culture of bivalves. Total organic carbon, total nitrogen, Eh values and pheopigments were significantly higher under the trestles than in any other stations. Other stations often did not differ from the reference station. 8. The effects of shellfish culture on the water column were. However, it was observed a small decrease of the food available to the molluscs near the rearing 9. The DEB model was able to describe and predict adequately the growth of oysters, both in the Baie des Veys and in the Loch Creran. The parametres for its use in other environment are given, but a tuning of one parametre should be performed with the help of authors. 10. Among the indicators and models for use in are as of intertidal bivalve culture, it is recommended to use the sediment quality index, TOC (Total Organic Carbon), redox and pheopigments, in surficial sediment, AMBI for the macrofauna, chlorophyll a contents and nitrogen forms in the water column, and models describing the carrying capacity, filtration rate of molluscs, and a DEB model to predict the growth of molluscs.