Annual/Termination Reports:

[05/01/2007] [06/20/2008] [06/26/2009] [04/20/2010]

Report Information

Participants

see meeting minutes for list of participants

Brief Summary of Minutes

Next year's meeting will take place in conjunction with the NSA meeting in Providence, RI. Marta Gomez-Chiarri will be chair for the meeting next year.

Station Reports

Kim Reece VIMS: Bivalve Genetic Studies
Genetics of the disease organisms and genetics of bivalves
" Even if you see genetic structure, it is a huge jump (especially given the history of stocking in CB) to say that populations are locally adapted.
" Developing molecular genetic markers focusing on three bivalve species

Ryan Carnegie VIMS Shellfish Pathology Laboratory
Health management in wild and cultured shellfish populations and oyster conservation with a focus on Haplosporiduim nelsoni (MSX disease) and Perkinsus marinus (dermo) in C. virginica

Ximing Guo: Oyster Genetics and Breeding at Rutgers
Selective Breeding, e.g., Cross-breeding of selected strains (Rawson)
Cytogenetics (evaluation of interstrain triploids, flurescence in situ hybridization)
Molecular Genetics and Genomics
Interstrain triploids versus diploid hybrids

University of Maine / Rawson
Concerns regarding inbreeding led to interline hybrids and timing of deployment
Controversy around who gets extra seed&one hatchery to another or the hatchery to growers.

Marta Gomez-Chiarri University of Rhode Island : Immune defenses in oysters
Goal: find mechanisms of disease resistance in oysters and host-pathogen in oysters

John Scarpa: Overview of the HBOI Molluscan Research Program (Fort Pierce, Fl)
" Shellfish retraining programs:
" Enhancing Production of Cultured Hard Clams (Mercenaria mercenaria) in Florida
" Sunray venus clams: new species to diversify the Florida hard clam aquaculture industry
" Conch Heritage Network: The Queen Conch is a species of concern
" Aquatic animal health laboratory: diagnostic and analytical services
" Harbor Branch Clams, Inc: Produce approximately 30% of clam seed for Florida and also supplied Georgia, Carolinas and Virginia


Molluscan Broodstock Program/ OSU (Ford Evans and Chris Langdon)
In 2004, implemented a rotational breeding program to control the rate of inbreeding within lines. But we may be setting ourselves up for outbreeding depression, or breaking up some of the beneficial gene clusters of these very successful lines.

Also investigating post-harvest oyster quality in Alaska and using a cooperative effort among OSU, Alaska Sea Grant, Fishery Industrial Technology Center to look at an industry based on the half-shell market

Aquaculture Genetics and Breeding Technology Center, VIMS Stan Allen
SOS production in Virginia has increased from under 10 million in 2005 to 100 million projected in 2007.
Looked at two basic approaches for environment by genotype interactions
Need to analyze the contribution of diseases to mortality line by line---as the Lousisana stocks cant handle MSX, die quickly and skew the results.

Joth Davis: Taylor Resources Hatchery
Bivalve Culture in Puget Sound, mostly introduced: Pacific oyster, Kumamoto oysters, European oyster, Olympia oyster, Manilla clam, Mussel, geoduck
Original WRAC project Objectives:
Research Component: Five objectives for improving the efficiency of testing the crossbreeding potential of inbred lines
Many goals: but will focus today on production and testing the performance of hybrid oysters (TRI)
Major Products
§ Evidence for growth heterosis
§ Physiological and genomic predictors of hybrid yield
§ A two tiered hybrid testing program
§ Methods and protocols for testing yield

Jason Curole: Update from Hedgecock lab
Larval heterosis: consistently observe heterosis at the larval stage and at the adult stage
Larval size distribution
ODRP
Looking at summer mortality
Kumamoto
GenEn
USDA (finished)
Fellowship (Jason Curole)
Seeing a maternal effect on mitochondrial expression

JGI: Oyster Genomics Work
Jason Curole (Hedgecock)
JGI Oyster Projecct
Paired-end sequencing of 150,000 cDNAs
Preliminary results from OX looked good so JGI went for a full production run.
Alas, those did not look as good.

Conversation about Summer Mortality
But Dan and Pierre are still out due to weather, so Joth presenting
Key findings:
19-20 degree sustained temperatures trigger mortality in France and along the West Coast
Time and time again, mortality comes back to hitting this 20 degree threshold
In some cases, Lower mortalities observed in MBP lines but it is not consistent
51x35 and 35x51 got wiped out in 2005

Other parameters
" Dissolved Oxygen: DO can be quite low later in the month of September that may be associated with mortality events. DO related directly to tidal exchange
" Much higher mortality when oysters are cultured on-bottom
" Much higher mortality at 30cm off bottom than at 70 cm off bottom
" No Ploidy influence
" Phytoplaknton:
o In Willapa bay, diatoms abundant with low dinoflagellate numbers. Ceratium and Pseudonitzschia were most abundant toxic samples
o In Totten Inlet: low levels of diatoms with high dinoflagellates. Selected taxa dominated by Ceratium, but toxic species were uncommon
" Systemic Bacterial Infections
" Vibrio and herpes clearly indicated in larval and juvenile mortalities in CA
" Asymetrical shell growth
o Triploid oysters show overgrowth of flat shell by cupped shell
o About 30% of samples with multifocal necrosis of gill tissues
o Pathology possible due to overheating and exposure during low tide

Current Research
" Test the hypothesis that small number of genes control resistance to summer mortality through genomic mapping of variation in survival, growth, and reproductive allocation
" Breeding program focused on selecting for resistance to summer mortality, to determine how variation in reproductive allocation may interact with stress conditions associated with summer mortality
" To evaluate selected environmental parameters and observations which appear to predict increased summer mortality risk and can be used to assess the performance of outplanted treatment groups
" To ensure that there is a timely response to oyster mortality events on commercial oyster beds, and harvest areas and provide feedback on those events to producers.


Addendum WERA99
Tuesday, February 27, 2007, 5:00-6:00pm

This mini meeting was held to continue the discussion started on Sunday with members of the group that were unable to make it over the weekend due to travel problems.


Addendum WERA99
Tuesday, February 27, 2007, 5:00-6:00pm

This mini meeting was held to continue the discussion started on Sunday with members of the group that were unable to make it over the weekend due to travel problems.

At the conference in Korea, Jean-Francois Samain had a powerpoint presentation that he used to review the summer mortality project (MOREST) in France. There was a question to Pierre Boudry about being able to get a copy of this presentation for the group. Pierre responded that he will ask Jean-Francois for a copy.

Pierre Boudry Oyster Genome Update from France
Following the application to JGI, Pierre applied for a BAC sequencing project, with the idea that it would be complementary. The problem was that it wasnt the same materialbut maybe it would help in some ways and not overlap between France and US.

This project was accepted conditional to the JGI projectso nothing could be done.

Then US did the EST the project and Genoscope (France) accepted sequencing of ESTs too. They did roughly the same thing but most of the cDNAs are different than what was done in the USso some overlap but not much.

One of the main differences is that this is based on normalized libraries
1) Immosite from colleagues from Montpeliar. This is done and in Paris ready to be sequenced
2) One on larvae which has not been completed
3) One on gonadhas been completed and ready to be sequenced. Not publicly available yet but will be at some point (6 months or so)

Big question: whether we go for the whole genome sequencing, what will be the best strategy with science and technical aspects AND the political side of the story.

Looks like China is ready to do the workthey want 3 million dollars to do the work in six months. But the idea of just providing money to have the work done is not very attractive to Francesame for US?

Following the step of sequencing, this is just the beginning of the story, a lot to be done afterwardsmaybe US, France and so on could offer to do the annotation or share the work among different sequencing platforms or do the work based on BAC libraries as it was done in many species until now. Best solution in terms of efficiency? In terms of politics?

Sequencing is really just the beginning. How would we even get it assembled? I dont know that any of us have the technical expertise or the computing power to do this. The idea of the workshop is to bring people in who have the political and the technical experience to do this: sequencing, assembly, annotation.

In 1-2 years time we can hope that we will have enough EST sequences because there are 2 projects in Europe plus the Hedgecock project.
Start with 30,000 hope to get 100,000 and depending on redundancy of sequences, will continue or not, plus the 150,000 from USA&.we get a massive amount of data and maybe a launching pad to get the genome sequenced.

Listserve specifically dedicated to this objective so that everyone can be up to date on what is going on? Very small number of emails that you would recieved but everyone could be kept abreast of what is going on. Could build from the WERA list.

Clear that whole story was driven by US but it looks like the idea is now to move to a more international communitydont know if we can accomplish everything in Europe, but genoscope considers oyster an important species. Wherever the work is done at the end is not importantbetter to work together internationally and get this done.

OGC: steering committee of the oyster genome consortium is talking about this international effort. International workshop is the place to start.
Volkswagen is paying to sequence the horse genome. BAC based.

Tom Coaker has convinced NIH to sequence the Tilapia genome. Very realistic, can come from anywhere, leave our options open.

List of all the genomes that have been done and a list of all the applied projects (like ours) that are on the way&.how they are organized, how theyve been done, what are the options to join forces between countries, etc.

Shrimp genome is an international effortkind of an impetus for OGC. Salmon and trout communities are very well organized and bigbut very useful to see how these groups have organized and advocated to have it done.

We should have some people from the other aquaculture groups whove gotten their genomes sequenced so that we can follow their lead.

Pierre will see if he can get some money from France for this too. The sooner the better.

Broad and quick update from France from Pierre
The moras project is finished we are currently working on a book that will summarize everything that was done. John Francoise is working very hard on itand book will be both in French and English

French research groups working on oysters:
Immunology and defense mechanisms (Montpellier)
Physiology (Brest and Caen)
Genetics (La Tremblade, Ifremer)

SNP identification in C. gigas (C. Sauvage)
Analysis of codon usage (8800 ESTs) (C. Sauvage)
QTL mapping of resistance to summer mortality in 5 F2 families
Functional genomics of R and S selected lines
Development oyster cDNA microarrays
Gene Expression Profiling
Analysis and Comparison at the levels

In Montpelier Ana Tirape worked with Viviane Boulo on immune related genes during ontogenesis

Antimicrobial defense mechanisms (montelier)

Genome evolution and speciation in closely related bivalve taxa

Have been working to implement a breeding program with some of the oyster farmers in Francenot easy because it is not in their habit to develop these kind of things but is one of the results of Morest. They use the example of MBP that farmers in France should get together to do breeding. Pierre thinks he should leave genetics and go to sociology nowpolitically, it has been very difficult.

Accomplishments

WERA-099 provided a forum for US and international molluscan geneticists, physiologists and pathologists to exchange ideas and information on genetics, reproduction, diseases, chromosome and genetic manipulation techniques, broodstock management and breeding programs. <br /> <br /> Multi-disciplinary and inter-regional exchanges among participants revealed new directions of scientific enquiry and facilitated formulation of new research questions. The committee helped overcome problems associated with geographical isolation facing many researchers located at remote marine field stations.<br /> <br /> Presentations by Dr. Pierre Boudry, IFREMER, France, provided US researchers with information on a large-scale oyster genetics program in France and discussion of international efforts to sequence the genome of the Pacific oyster. <br />

Publications

2006 Publications<br /> <br /> Matson, S., Evans, F. and C. Langdon. 2006 Specific pathogen-free culture of the Pacific oyster (Crassostrea gigas) in a breeding research program: effect of water treatment on growth and survival. Aquaculture. 253: 475-484.<br /> Evans, S. and C. Langdon. Effect of dietary restriction during juvenile development on adult performance of Pacific oysters (Crassostrea gigas). Aquaculture, accepted<br /> <br /> Evans, S. and C. Langdon. Effects of genotype x environment interactions on the selection of broadly adapted Pacific oysters (Crassostrea gigas). Aquaculture, 261: 522-534.<br /> <br /> Evans, S. and C. Langdon. 2006. Direct and correlated responses to selection for individual body weight in the Pacific oyster (Crassostrea gigas). Aquaculture, 261: 546-555.<br /> <br /> Delaporte, M, Chu F.L, Langdon, C, Moal , J, Lambert, C, Samain, J-F and Soudant, P. 2007. Changes in biochemical and hemocyte parameters of the Pacific oysters Crassostrea gigas fed T-Iso supplemented with lipid emulsions rich in 20:5n-3. J. Experimental Marine Biology and Ecology, 343:261-275.<br /> <br />

Impact Statements

1. Restoration of the Chesapeake Bay with native or introduced oyster species or both is an important national goal. Presentations by several east coast researchers highlighted some of the major problems and possible solutions in meeting this objective. Conversations between researchers at VIMS and MBP should result in the development of a more efficient breeding program to produce disease-resistant strains.
2. US and French efforts to map the genome of the Pacific and Eastern oyster and to sequence the genome of the Pacific oyster are moving forward. Exchange of information on the development of new maps and mapping techniques will lead to a better understanding of the genetic basis of disease resistance in Eastern oysters as well as growth and survival in farmed Pacific oysters.
3. see minutes for additional points

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Report Information

Participants

Allen, Stan, ska@vims.edu,Virginia Institute of Marine Sciences; Auyong, Jan,jan.auyong@oregonstate.edu,Oregon State University,OR; Camara, Mark,mark.camara@oregonstate.edu,USDA ARS - Oregon,OR; Cordes, Jan,jfcordes@vims.edu,Virginia Institute of Marine Sciences,VA; Evans, Judd,juddevans@activ8.net.au,South Australian Oyster Growers Association,AUSTRALIA; Frank-Lawale, Anu,afl@vims.edu,Virginia Institute of Marine Sciences,VA; Gaffney, Patrick,pgaffney@udel.edu,University of Delaware,DE; Gomez-Chiarri, Marta,gomezchi@uri.edu,University of Rhode Island,RI; Guo, Ximing,xguo@hsrl.rutgers.edu,Rutgers University,NJ; Hedgecock, Dennis,dhedge@usc.edu,University Southern California,CA; Jensen, Gary,gjensen@csvees.usda.gov,USDA CSREES,DC; Langdon, Chris,chris.langdon@oregonstate.edu,Oregon State University,OR; Leavitt,Dale,dleavitt@rwu.edu,Roger Williams University,RI; Lindell, Scott,slindell@mbl.edu,Marine Biological Laboratories,MA; Matson, Sean,matsonse@onid.orst.edu,Oregon State University,OR; Milbury, Corey,cmilbury@hsrl.rutgers.edu,Rutgers University,NJ; Plough, Louis,lplough@usc.edu,University Southern California,CA; Rawson, Paul,prawson@maine.edu,University of Maine,ME; Reece, Kim,kreece@vims.edu,Virginia Institute of Marine Sciences,VA; Roberts, Steven,sr320@u.washington.edu,University of Washington,WA; Silkes, Bill,bsilkes@americanmussel.com,American Mussel,RI; Straus, Kristi,kmstraus@u.washington.edu,University of Washington,WA; Troup, Tony,troup@tsn.cc,Camden Haven Oyster Supply,AUSTRALIA; Varney, Robin,rvarney@udel.edu,University of Delaware,DE; Wang, Yan,yan@hsrl.rutgers.edu,Rutgers University,NJ; Wheaton, Fred,fwheaton@umd.edu,Northeastern Regional Aquaculture Center,MD; Zhang, Liusuo ,lzhang@hsrl.rutgers.edu,Rutgers University,NJ.

Brief Summary of Minutes

The 2009 annual meeting will take place in conjunction with the NSA meeting in Savannah, Georgia. Dr. Frank-Lawale will be chair for the 2009 meeting.
Station Reports


University of Maine and Marine Biological Laboratories: Paul Rawson reported on a collaborative project between institutions in the Northeast and oyster farmers, on the crossbreeding and field trials for disease resistant oysters. Two oyster lines were crossed, the MSX and Dermo-resistant NEH and the University of Maine/Industry Cooperative Program UM Flowers select (UMFS), with putative resistance to Juvenile Oyster Disease. Parental lines, a UMFS x NEH F1 hybrid, as well as two backcrosses (NEH F1 x UMFS and NEH F1 x NEH) were deployed in locations in the Northeast and tested for performance. The hybrid line (UMFS x NEH) showed fastest growth at Rhode Island, Connecticut, and Massachusetts sites through the first season. The backcross line F1 X NEH has relatively high survival at sites where Dermo disease (New Jersey), MSX disease (Connecticut and Massachusetts) and Juvenile oyster disease (Rhode Island) occur with no apparent difference in growth performance. Although results are only preliminary and disease testing needs to be performed, the results from the first season indicate the promise of using hybrid or backcross lines for enhanced resistance to multiple diseases.


University of Rhode Island. The Gomez-Chiarri laboratory has been working on the evaluation of the performance of three oyster lines/stocks (Rutgers NEHY, Frank M. Flowers from Long Island, and a local stock from Green Hill Pond in Rhode Island) in Rhode Island farms. The FMF stock showed the highest mortality in both field and experimental challenges. No statistical significances were seen in performance between the stocks, due to variations in performance at the different farms. Mortality happened throughout the year and dermo disease prevalence and intensity were low in both farms. A hybrid between the NEHY line and the local stock from Green Hill Pond will be produced in 2008 and performance tested in several farms. A second project, related to the search of markers for disease resistance, involves the investigation of the role of a candidate molecule, a matrix metalloproteinase (MMP) from the Eastern oyster, in immune responses. Tissue patterns of production of MMP in oysters showed that the protein is induced in mucosal surfaces and hemocytes in response to infectious challenge, feeding, and shell damage, suggesting potential roles in immune defenses, shell formation, and digestion.


Rutgers University. Ximing Guo reported on the selective breeding efforts at Rutgers University. The line NEHY, selected for 48 years for disease resistance, shows strong resistance to MSX disease and some resistance to dermo disease. A hybrid between NEHY and the Flowers line (resistant to juvenile oyster disease - JOD) shows improved yield for the hybrid compared to parental lines and improved resistance to JOD in field trials. Triploids of NEHY and the NEHY x Flowers hybrid tested in field conditions in NJ and MA showed improved resistance to JOD and dermo disease and higher yield. Triploids outperformed diploids, and hybrid outperformed pure stock, showing that the selective advantage in diploid line gets transferred to the triploid line. Triploids may have performed better against JOD because got to refuge size (more than 25 mm in shell height) faster. Dr. Guo also reported on the progress of the projects on the mapping of genes involved in disease resistance. Markers developed by the Rutgers laboratory is up to 239 microsatellite loci (93 genomic, 146 from ESTs) and 102 SNPs from ESTs, most corresponding to genes involved in host defense (over 300 co-dominant markers). The current resolution of the linkage maps is 2-6 cM. Oyster families have been developed to map genes involved in disease resistance by family-based association. Several QTLs that may be involved in resistance to dermo disease have been identified. The laboratory is now developing 81 families (9 x 9 crosses) for marker-assisted breeding.



University of Delaware. Pat Gaffney reported on the global population genetic structure in Crassostrea virginica. Single nucleotide polymorphism (72 loci) allele frequency variation was detected across the species range, showing large scale structure (Gulf, S Atlantic, N. Atlantic) and localized heterogeneity/ structure. Null alleles are common. Primers for the amplification of a few SNP markers have been developed that work in different oyster species. A bacterial artificial chromosome (BAC) library (from an individual, highly inbred C. gigas oyster) has been constructed and has been fingerprinted. The resulting physical map will be combined with genetic maps, allowing for targeted analysis of selected genes and selection of evenly spaced high-density markers for QTL mapping. The library can also be used to facilitate a whole genome sequencing project.


Virginia Institute of Marine Sciences (VIMS) I. Kim Reece reported in the development or genetic markers for Crassotrea virginica, Crassostrea ariakensis, and Mercenaria mercenaria, and on the application of the markers in different project. Genetic markers for C. virginica, which allow distinguishing between the DEBY line (used in this restoration project) and local wild oyster stocks have been used to track the success of restoration projects in Chesapeake Bay. Data suggested that DEBY contribution has been low, maybe due to predation, poor survival and/or poor reproduction. Large deployments of DEBY seed (12 million) were performed in 2005-07 with efforts at protection of the seed with netting. However, genetic signal from DEBY line at the restoration site is still not above background. Models indicate that the lack of genetic signal may be due to physical characteristics of the site leading to low larval retention. In another project, the utility of 8 microsatellite loci for evaluating hatchery-selected lines of Crassostrea virginica was tested. The markers were able to discriminate among 4 hatchery lines and retrospectively quantify the success of the various crosses involving 1, 2 or more of the lines. Self assignment tests showed greater than 95% assignment for all parental lines. Markers for C. ariakensis were used to inventory germplasm resources. Genetic variation and differentiation among natural populations of C. ariakensis from China, Korea, and Japan was examined, and compared to hatchery stocks in the US. Studies showed temporal stability and population structure (isolation by distance) in natural Asian populations. A reduction in expected heterozygosity and allelic richness was observed in older hatchery stocks of C. ariakensis in the US, while in new stocks (a single generation in hatchery) no significant differences in heterozygosity but a reduction in allelic richness were observed. Eight molecular markers for hard clams, Mercenaria mercenaria, have been developed and tested, and show overall significant difference between Virginia and South Carolina wild populations.


VIMS (II). Stan Allen reported in the Aquaculture and Breeding Technology Center. The goal of the center is to provide shellfish broodstock to industry. A total of 11 lines have been developed, including DEBYs and Crosbreed (XB), as well as several Louisiana-derived lines and crosses between the three. Lines are tested in several locations and best performers are saved for broodstock (mass selection). Results indicate a strong interaction between genetics and environment, warranting the development of regional varieties. A line (WTS) derived from natural pops of Chesapeake Bay oysters that might have developed resistance to dermo disease has been developed.


University of Washington (I). Kristi Straus reported for the Friedman laboratory. Mitochondrial and nuclear markers indicate that a population of abalone from the San Juan Islands (WA) that visually look like pinto abalone is more genetically similar to flat abalone. The biological relevance of these observations is being evaluated in fertilization studies. Another project plans to evaluate the effect of geoduck aquaculture on wild populations, based on the observation that aquaculture geoducks show high rates of maturation and spawning. The Friedman laboratory is also developing tools for the diagnosis of oyster herpes virus and abalone withering syndrome using quantitative real time PCR. These tools are being used in projects related to the development and evaluation of disease resistance strains. A novel Ricketssia-like bacterium has been detected in abalone, as determined by morphology, susceptibility to antibiotics, and genetic analysis.


University of Washington (II). Steven Roberts reported on the development of a real time PCR method for the quantification of Quahog Parasite Unknown in hard clams. He also reported on the identification of molecules involved in disease resistance/ tolerance in different species of shellfish. Genes potentially involved in immunity in oysters have been identified from a EST library from oyster granulocytes. One of the genes is a homolog from the cytokine interleukin 17. A stock of oysters derived from survivors of a dermo disease outbreak in Marthas Vineyard, MA, has been developed and tested in the field in MA. The stock showed improved performance. Differences in patterns of expression of constitutive genes were observed between the resistant and a susceptible stock. The Roberts laboratory, in collaboration with the Friedman laboratory, is starting to mine ESTs for abalone that may be involved in resistance to Withering syndrome. The Roberts lab is also looking at differences in gene expression in lines of hard clams showing differences in resistance to QPX disease.


Molluscan Broodstock Program (MBP). Chris Langdon reported on the search for Kumomoto oyster broodstock from Japan that is not inbred and has not been hybridized with the Pacific oyster, C. gigas. The search reveled that there is no species-specific shell morphology for Kumamoto oysters that would allow them to be differentiated from C. gigas, and points to the need of genetic markers that allow to differentiate between different oyster species. Currently available genetic markers do not distinguish between C. angulata and C. gigas. Dr. Langdon also reported on the breeding program. As of 2006, the MBP have produced about 1200 families from 7 original broodstock lines. The MBP relies now on rotational breeding to avoid inbreeding. The response to selection after two generations indicates that, on average, the top 5% are 70% better than progeny from unselected broodstock. Improvements in yield in the Pipestem line are related to improved survival, and this line is being considered for commercial production. MBP is now generating inbred parental lines using an Adam and Eve strategy of crosses for family amplification which show good resistance to summer mortality. Another line of research relates to selection for color pattern in Pacific oysters. The program surveyed 20+ people at two locations and found that people generally prefer mottled or stripey oysters instead of pure black or white. Overall color of the shell is highly heritable and could be manipulated through breeding
In most familes there is a continous optical density distribution. But in some families you see a bimodal distribution with about 3:1 ratio or a 1: 1 ratioindicating a single major gene controlling shell color. In collaboration with Camara and Lang, the MBP is searching for genetic markers for marker assisted selection using microarrays to identify differences in ESTs between sensitive and tolerant MBP families to stress (acute heat-shock). The ESTs are verified using qPCR, and differences in expression are correlated with field performance traits. About 20 ESTs are correlated with either oyster growth in field or yield (i.e. galectin).


USDA ARS Shellfish Genetics Program (Oregon). Mark Camara reported on several ongoing projects. On the first project, regarding the importation and evaluation of new Kumamoto oyster germplasm to address problems with inbreeding and hybridization, two sets of markers allowed identifying pure Kumomoto oysters and hybrids with other oyster species in the Pacific, indicating that while there is no evidence for extensive hybridization in the Ariake Sea, hybridization barriers appear to be lower in US cultured oysters. Furthermore, morphology cannot be reliable used to distinguish between Pacific and Kumomoto oysters. The ARS lab is also involved in the genetic characterization of existing Pacific oyster germplasm resources using AFLP markers. On a third project, the ARS lab has developed software tools to optimize pedigree reconstruction using molecular markers & mixed-family selection. The lab has also identified candidate genes for stress resistance in Pacific oysters using DNA microarrays, cDNA-AFLP differential display, and RT-qPCR, is mapping quantitative trait loci for economically important traits in Pacific oysters; and is characterizing molecular genetic variation within and among remnant populations of native Olympia oysters.



University of Southern California. Dennis Hedgecock reported on the centromere mapping for the Pacific oyster, Crassostrea gigas. Expectations from cytology indicate that the Pacific oyster has 10 linkage groups and ~600 cM map units, but linkage studies suggest that the genome is twice as big. Gene centromere mapping was performed based on segregation in chemically-induced triploids (c3N). That most markers (87/93) show 1:1 homozygote ratios in c3N is consistent with the likelihood that paternal allele in these triploid offspring mask the effects of "homozygosity" for any recessive mutations coming from the female. There are also indications of chromosomal rearrangements. Six out of 10 centromeres were marked, facilitating mapping of new markers. Interference is much less than previously believed; explaining why genome size estimated in linkage studies is larger than cytological estimations.


International Participation: Shellfish Aquaculture in Australia. Judd Evans and Tony Troup, two farmers sponsored by Australian Seafood Cooperative Research Centre reported on oyster aquaculture industry and research in Australia. There are three separate oyster industries in Australia (Tasmania, New South Wales and Southern Australia), hoping to combine into one cohesive industry. The Australian oyster industry has US 1 billion, 1300 direct jobs, and 3000 indirect jobs invested in the production of Sidney rock oysters, flat oyster, and C. gigas. Research projects include a C. gigas breeding program, which began in 1997 (CSIRO) and was handed to industry in 2001. The selectively bred oysters showed improved growth rate (5% increased growth per generation) at the expense of changes in shape, so the program now selects for increased survivability, shape, and growth rates. The SRO breeding program has developed lines resistant to the disease QX (caused by Marteilia sydneyi) that show improvements in growth and survival. Further breeding development by the Sidney Oyster Company (SOCo) is geared to increased growth, condition, shape, and survival to QX and winter mortality. Efforts also include the development of hatchery production of selectively bred stock, creation of a nursery network, and environmental awareness, best management practices, and quality controls. Current research on QX disease (Marteilia sydneyi) has led to the development of markers for QX resistance, diagnostic probes, and identification of 2 polychaete species as the primary host for the parasite.


Gary Jensen USDA CSREES, and Fred Wheaton, Northeast Regional Aquaculture Center, reported on grant opportunities, Dr. Jensen encouraged researchers to consider applying to integrative programs.


Oyster Genomics Update (Hedgecock, Gaffney). JGI is moving forward on the EST sequencing project of cDNA libraries that were developed from C. gigas inbred lines 35 and 51. The sequencing of BACs that had some genes of physiological interest is close to being complete. The first BAC that was done had 9-10 genes including cytochrome P450. An annotation workshop will be scheduled sometime in the Fall. Next steps should include: 1) placing markers onto the map; 2) find funding for BAC end sequencing to construct a high quality map


Broodstock for restoration projects. The use of selectively bred stocks for restoration projects was briefly discussed, projects should strive to maintain local adaptation and avoid bottlenecks. Issues to consider include number of broodstock, effective population size, etc. The idea of so called genetic restoration has been reconsidered in the favor of trying to persist with wild stocks that may show evidence of development of disease resistance, combined with good hatchery techniques.


C. ariakensis update. Environmental impact statement is still being crafted due in May. Nothing is really happening except an occasional deployment of a million or so triploids.

Accomplishments

WERA-099 provided a forum for US and international molluscan geneticists, physiologists and pathologists to exchange ideas and information on genetics, reproduction, diseases, chromosome and genetic manipulation techniques, broodstock management and breeding programs. Highlights of the 2008 meeting included: 1) increased knowledge about the relative performance of selectively bred strains, hybrids, and triploids in farms in the Northeast US; 2) intense efforts towards marker development; 3) development of molecular diagnostic tools for diseases affecting the bivalve industry; 4) efforts towards the identification of molecules involved in bivalve immune responses and resistance to stress; and 5) progress towards mapping and genome sequencing. <br /> <br /> <br /> Multi-disciplinary and inter-regional exchanges among participants revealed new directions of scientific enquiry and facilitated formulation of new research questions. The meeting also provided a forum to stimulate collaborations between US and Australia. <br />

Publications

Batista FM, Arzul I, Pepin JF, Ruano F, Friedman CS, Boudry P, Renault T (2007) Detection of ostreid herpesvirus 1 DNA by PCR in bivalve molluscs: a critical review. J Virol.Methods 139:1-11<br /> <p><br /> Burge CA, Judah LR, Conquest LL, Griffin FJ, Cheney DP, Suhrbier A, Vadopalas B, Olev PG, Renault T, Friedman CS (2007) Summer seed mortality of the Pacific oyster, Crassostrea gigas thunberg grown in Tomales Bay, California, USA: The influence of oyster stock, planting time, pathogens, and environmental stressors. J.Shellfish Res 26(1):163-172<br /> <p><br /> Camara MD, Evans S, Langdon C (2007) Inbreeding effects on growth and survival in a naturalized population of the Pacific oyster (Crassostrea gigas) revealed using molecular marker-based estimates of parental relatedness. Aquaculture 272:S247<br /> <p><br /> Curole JP, Hedgecock D (2007) Bivalve Genomics: Complications, Challenges, and Future Perspectives. Aquaculture Genome Technologies. Jul 2007, pp 525-543.<br /> <p><br /> Gomez-Leon J, Villamill L, Salger SA, Sallum RH, Remacha-Trivino A, Leavitt DF, Gomez-Chiarri M (2008) Survival of eastern oysters Crassostrea virginica from three lines following experimental challenge with bacterial pathogens. Dis Aquat Organ 79:95-105<br /> <p><br /> Hedgecock D, Davis JP (2007) Heterosis for yield and crossbreeding of the Pacific oyster Crassostrea gigas. Aquaculture 272:S17-S29<br /> <p><br /> Hedgecock D, Launey S, Pudovkin AI, Naciri Y, Lapegue S, Bonhomme F (2007) Small effective number of parents (Ndb) inferred for a naturally spawned cohort of juvenile European flat oysters Ostrea edulis. Mar.Biol. 150(6):1173-1182<br /> <p><br /> Hedgecock D, Lin JZ, DeCola S, Haudenschild CD, Meyer E, Manahan DT, Bowen B (2007) Transcriptomic analysis of growth heterosis in larval Pacific oysters (Crassostrea gigas). Proc.Natl Acad.Sci U S.A 104:2313-2318<br /> <p><br /> Hedgecock D, Perry GML, Voigt ML (2007) Mapping heterosis QTL in the Pacific oyster Crassostrea gigas. Aquaculture 272:S267-S268<br /> <p><br /> Jenny MJ, Chapman RW, Mancia A, Chen YA, McKillen DJ, Trent H, Lang P, Escoubas JM, Bachere E, Boulo V, Liu ZJ, Gross PS, Cunningham C, Cupit PM, Tanguy A, Guo X, Moraga D, Boutet I, Huvet A, De GS, Almeida JS, Warr GW (2007) A cDNA microarray for Crassostrea virginica and C. gigas. <br /> Mar Biotechnol.(NY) 9:577-591<br /> <p><br /> Moss JA, Burreson EM, Cordes JF, Dungan CF, Brown GD, Wang A, Wu X, Reece KS (2007) Pathogens in Crassostrea ariakensis and other Asian oyster species: implications for non-native oyster introduction to Chesapeake Bay. Dis Aquat Organ 77:207-223<br /> <p><br /> Moss JA, Xiao J, Dungan CF, Reece KS (2008) Description of Perkinsus beihaiensis n. sp., a new Perkinsus sp. parasite in oysters of Southern China. J Eukaryot.Microbiol 55:117-130<br /> <p><br /> Quilang J, Wang S, Li P, Abernathy J, Peatman E, Wang Y, Wang L, Shi Y, Wallace R, Guo X, Liu Z (2007) Generation and analysis of ESTs from the eastern oyster, Crassostrea virginica Gmelin and identification of microsatellite and SNP markers. BMC Genomics 8:157<br /> <p><br /> Roberts S, Gueguen Y, de LJ, Goetz F (2008) Rapid accumulation of an interleukin 17 homolog transcript in Crassostrea gigas hemocytes following bacterial exposure. Dev Comp Immunol 32:1099-1104<br /> <p><br /> Taris N, Lang RP, Camara MD (2008) Sequence polymorphism can produce serious artefacts in real-time PCR assays: hard lessons from Pacific oysters. BMC Genomics 9:234<br /> <p><br /> Wang Y, Guo X (2007) Development and characterization of EST-SSR markers in the eastern oyster Crassostrea virginica. Mar Biotechnol.(NY) 9:500-511<br />

Impact Statements

1. Conversations between researchers should result in the development of markers for marker-assisted selection that could be used in more efficient breeding programs to produce disease-resistant strains.
2. Research efforts are geared to provide industry with stocks, families, and lines that show improved performance.
3. International efforts to map the genome of the Pacific and Eastern oyster and to sequence the genome of the Pacific oyster are moving forward. Exchange of information on the development of new maps and mapping techniques will lead to a better understanding of the genetic basis of disease resistance in Eastern oysters as well as growth and survival in farmed Pacific oysters.

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Report Information

Participants

Allen, Stan, ska@vims.edu, Virginia Institute of Marine Sciences, VA; Auyong, Jan, jan.auyong@oregonstate.edu, Oregon State University, OR; Camara, Mark, mark.camara@oregonstate.edu, USDA ARS - Oregon, OR; Cordes, Jan, jfcordes@vims.edu, Virginia Institute of Marine Sciences, VA; Davis, Joth, jdavis@wolfenet.com, Taylor Shellfish Farm, WA; Frank-Lawale, Anu, afl@vims.edu, Virginia Institute of Marine Sciences, VA; Gomez-Chiarri, Marta, gomezchi@uri.edu, University of Rhode Island, RI; Guo, Ximing, xguo@hsrl.rutgers.edu, Rutgers University, NJ; Hedgecock, Dennis, dhedge@usc.edu, University Southern California, CA; Jensen, Gary, gjensen@csvees.usda.gov, USDA CSREES, DC; Kube, Peter, Peter.Kube@csiro.au, CSIRO, AUSTRALIA; Langdon, Chris, chris.langdon@oregonstate.edu, Oregon State University, OR; LaPeyre, Jerome, jlapeyre@agcenter.lsu.edu, Louisiana State University, LA; Proestou, Dina, proestou@uri.edu, University of Rhode Island, RI; Rawson, Paul, prawson@maine.edu, University of Maine, ME; Reece, Kim, kreece@vims.edu, Virginia Institute of Marine Sciences, VA; Roberts, Steven, sr320@u.washington.edu, University of Washington, WA; Scarpa, John, jscarpa1@hboi.fau.edu, Harbor Branch FL Atlantic University, FL; Silverstein, Jeff, Jeff.Silverstein@ARS.USDA.GOV, ARS, USDA, DC; Spikers, Tom, Shellfish Culture, AUSTRALIA; Stick, David, david.stick@oregonstate.edu, Oregon State University, OR; Wilbur, Ami, wilbura@unc.edu, University of North Carolina, Wilmington, NC

Brief Summary of Minutes

Brief Summary of Minutes of Annual Meeting:
Station reports were brief at this meeting in order to make time for group discussions and a presentation on the application of quantitative genetic principles to oyster breeding by Dr. Peter Kube from CSIRO, Australia. The 2010 annual meeting will take place in conjunction with the NSA meeting in San Diego, California. Dr. Steven Roberts will be chair for the 2009 meeting assisted by Dr. Allen.

Station Reports
University of Maine and Marine Biological Laboratories. Paul Rawson reported on the completed collaborative project between institutions in the Northeast and oyster farmers, on the crossbreeding and field trials for disease resistant oysters. Two oyster lines were crossed, the MSX and Dermo-resistant NEH and the University of Maine/Industry Cooperative Program UM Flowers select (UMFS), with putative resistance to Juvenile Oyster Disease. Parental lines, a UMFS x NEH F1 hybrid, as well as two backcrosses (NEH F1 x UMFS and NEH F1 x NEH) were deployed in locations in the Northeast and tested for growth and survival. There appeared to be a significant site x genotype interactions in mortality. Mortality was higher for NEH and NEH hybrid in all three Maine sites whereas almost all the UMFS animals died at New Jersey site. Growth did not vary among lines within sites. However, it did vary between Maine sites and New Jersey site with performance lower at the New Jersey site. The hybrid line (UMFS x NEH) exhibited highest performance in growth and survival at Maine sites

University of Rhode Island. The Gomez-Chiarri laboratory also reported on the evaluation of oyster lines in Rhode Island. They presented results from the testing of three oyster lines (i-Rutgers NEH, ii-GHP a wild stock from Green Hill Pond in Rhode Island where disease pressure is high and iii-a hybrid cross of the two) in 4 Rhode Island farms, two in Narragansett Bay and two in coastal ponds. The NEH line grew significantly better at the 2 Bay sites while the GHP line seemed to grow better in coastal ponds, however this was not statistically significant. By the end of summer, NEH line experienced significantly lower mortality than the two other lines at the Bay sites. The same was true at the coastal pond sites as well, but this was not significantly different. Overall mortality was significantly higher at one of the Narragansett Bay sites (site 2) for all lines. In order to determine the cause of mortality tests were done. Conchiolin was detected at the coastal ponds sites in late August, but very small percentage of oysters examined had it. It was also detected in both bay sites with and incidence level of 14% and 37% in both sites.
They are also gave a report on their project looking at the role of Matrix Metalloproteinases (MMPs) in innate immunity. They isolated a novel MMP in oysters-Cv1MMP and immunohistochemistry of this MMP shows that it is produced in epithelial barrier tissues (mantle, digestive) and hemocytes. An activity assay suggested that protease production increases in response to challenge with P. marinus. They have also found 41 SNPs in coding region of the Cv1MMP gene; on average 1 every 40 bp.

Rutgers University. Ximing Guo reported on several molecular and selective breeding projects at Rutgers University. The line NEH and the FMF line are now highly inbred therefore crossing them should result in some beneficial heterosis. When tested the cross between NEH and FMF resulted in a higher performing hybrid (with respect to yield) than either parental line. Triploids outperformed diploids and hybrid triploids from more than one line outperformed triploids of pure stock, suggesting that the selective advantage in diploid line gets transferred to the triploid line. His group has also identified 341 Co-dominant markers for C.virginica 239 SSRs (Simple sequence repeats) -93 from genomic, 146 from ESTs (express sequence Tags). In addition they have SNPs (Single Nucelotide polymorphisms) from 102 ESTs. They are using these markers to map genes for disease resistance using family-based association. The current resolution of the linkage maps is 1.15 cM and they have identified 5 chromosome regions associated with Dermo resistance using data from 3 families. There is now a marker assisted breeding project under way at Rutgers using 81 families from 3 lines (NEH, NEF and NEL) in a 9x9 matrix. Preliminary results show that the NEF line is the best. Dr. Guo also gave an update on the oyster genome project in China aimed at producing a draft sequence using next generation sequencing (50X Solexa pair-end reads, additional 454 coverage if needed). Results so far indicate that there are too many SNPs (even in 1 individual) and current computer programs cannot distinguish between the two chromosome copies for proper assembly.

Louisiana State University. Jerome LaPeyer made his debut presentation at the WERA099 meeting and gave a report on current oyster research in his lab at LSU. His lab focuses on mechanisms of disease resistance in the Louisiana (LA) oyster, which is primarily resistance to dermo. They compared survival between native LA oysters, the Crossbreed oyster (an MSX resistant line developed at Rutgers) and the native Chesapeake Bay oysters. Results showed that the LA oysters had lower parasite levels in their tissues suggesting that they are more resistant. There is now a small breeding population in their lab, now in their fourth generation, that has been selected for resistance to dermo. These animals are going to be compared with wild LA animals. In another experiment they looked at hemocyte activity in susceptible (Maine) and resistant (Louisiana) oysters. They observed a higher hemocyte density/proliferation in infected oysters and resistant (LA oysters). These oysters were able to clear infection better. They have also identified proteins (CvSI-1 and CvSI-2) that may be involved in response to Perkinsus, specifically proteinase inhibitors. They were found in plasma, epithelia, and digestive tubules of oysters. The expression level of CvSI-1 is higher in resistant oysters and thus the capacity to inhibit Perkinsus is much higher in selected oysters. The goal is to develop research tools (antibodies, clone isolation, recombinant, etc.) to study their role in response to infection

Virginia Institute of Marine Sciences (VIMS) I. Jan Cordes presented work on the genetic evaluation of recruitment success of deployed C. virginica on man-made reef in Chesapeake Bay conducted in the Reece Lab in VIMS. They deployed DEBY (a disease resistant line developed at VIMS) oysters on a shell Bar Reef and monitored recruitment using spat collectors. Spat were genotyped to distinguish between wild, DEBY and hybrid animals. They used two mitochondrial genes (COI and COIII); and only2% of oysters genotyped possessed DEBY haplotype. In 2006 and 2007 about 5% of spat can be attributed to DEBY-these years represented larger set of wild oysters as well. When microsatellite markers where used, 6% of spat were identified as DEBY in 2006 and 1% in 2007. These results pose questions about the success of planting domesticated animals in restoration exercises as no DEBY signals were found in neighboring sites either. Dr. Cordes also mentioned the advisory role that the lab plays. They deal with the verification of sighting of C. ariakensis in Virginia Rivers. So far they have confirmed two sightings- Coan River and York River-, but the animals were triploid and thus sterile.

VIMS (II). Stan Allen reported on the line performance and future direction of the line breeding programme at Aquaculture and Breeding Technology Center. The goal of the center is to provide genetically improved broodstock to industry. A total of 11 lines were developed, including DEBYs and Crossbreed (XB), as well as several Louisiana-derived lines and crosses between the three. Lines are tested at 4 different locations and best performers are saved for broodstock (mass selection). Results indicate selection for disease resistance has been successful; all best of lines perform better than wild control, however, there were no observed advantages over the control in size and weight at harvest. The total yields of lines were much higher than the control due to the highly significant difference in survival. These lines are now being coalesced into three main Super lines based on the origin of their parental line. These are DEBY, CROSSBREED and LOUISIANA. The Louisiana line has been diverged into high salinity (hANA) and low salinity (LOLA). Efforts are now going to be concentrated on improving growth rate. A new challenge for the center is how to produce and disseminate the large numbers of broodstock that the industry requires. A new breeding plan has been developed using larger numbers of animals to meet this need. Selection will also be done annually to produce odd and even year cohorts. As the industry develops new problems emerge and there appears to be an issue with low fecundity. This could be caused by chronic disease (dermo), pea crabs and inbreeding. The center also faces the challenge of conditioning the large number of broodstock needed to propagate the lines successfully. This new breeding and dissemination strategy will be online by 2010.

University of Washington. Steven Roberts reported on genetic studies of Vibrio tubiashi, a major problem for the shellfish industry. A disease challenge of C.gigas with Vibrio was conducted to identify genes involved in disease response (Interluekin, Prostaglandin 7 receptor). They identified ESTs containing microsatellites. Disease challenges were also conducted with three other shellfish species. A challenge with C. virginica yielded potential indicators using resistant and susceptible lines. Lab trials with H. cracherodii using resistant/susceptible suggest different levels of gene expression and raised a question about gene expression being used as a phenotypic marker for marker-assisted breeding? With M. mercenaria they observed indicators of resistance to QPX. They are sequencing the genome (short reads) to find some potential genes to look at, but they are currently using Sole and there may be problems with assembly.

Molluscan Broodstock Program (MBP). Chris Langdon reported on the problems faced by oyster hatcheries in the Pacific Northwest and MBPs role in helping the industry. Last year, larval production at hatcheries in the region was extremely low. They looked at conditions leading to lack of natural set of oysters in Willapa Bay and performed experiments in lab. They concluded that the source of the problems were Vibriosis, deceased PH and higher water temperatures. They increased water treatment capabilities in their hatchery (filtration, uv sterilization, and foam fractionation) and found that although better water quality improved larval growth the mortality still remained. They found that the lower pH in the ocean water decreased CaCO3 saturation levels and negatively affects larval shell deposition. Furthermore warmer water decreases solubility of dissolved oxygen by about 30%, resulting in lower larval survival. This problem was addressed by aeration.

Chris Langdon also gave an update on the performance of MBP families. The MBP adopted a rotational breeding design aimed preventing inbreeding. Results from cohort 18 from this scheme showed that the line was superior to wild and unselected controls in terms of yield (survival and body weight). Families were also tested for increased resistance to possible stressors in hatchery. Results from evaluation of families after two generations of selection suggested better performance relative to non-selected broodstock. The MBP has established a commercial repository to supply hatcheries with inbred parental lines. It is hoped that commercial hatcheries will use MBP lines for production and pass on their superior performance as seed to growers. So far the process has been extremely successful. Finally after two years of quarantine and $16,000 worth of disease testing Kumamoto oysters from Japan were seen to be free of diseases for 2 generations, and so have been planted in Yaquina Bay. There were no differences observed between Japan oysters and West Coast US oysters with respect to growth. This new Japan stock will be available through Taylor Shellfish.

USDA ARS Shellfish Genetics Program (Oregon). Mark Camara reported on several ongoing projects. On the first project he presented information on the population genetics of pacific oysters using material from Japan, New Zealand, Pacific Northwest and MBP lines. He used AFLP markers to generate a tree of the relationships between different stocks and found that the tree did not agree with history as the MBP populations sorted with Japan populations. In the second project he gave details of a mixed family selection strategy using molecular markers to retrospectively assign pedigree after pooling offspring from several parents in order to remove common environmental effects in the hatchery. The idea is to answer the question when is the best time to mix larvae? Straight hinge, pediveliger, d-larvae, post-larvae? Results were compared from mixing at each of the larval stages and it was found that earlier mixing resulted in higher genetic variability. In a third project he conducted heat tolerance test of all MBP families. The idea was to identify tolerant and susceptible families and correlate survival in the field to survival in the laboratory test. This was done to develop a laboratory test that could provide an indication of susceptibility to summer mortality. Further molecular tests using microarray profiling were done unfortunately, polymorphism lead to qPCR artifacts. An alternative approach using reverse microarray is being considered , however, it is not yet optimized. Dr. Camara then talked about future projects. His first project will be looking at QTLs for gene expression when summer mortality is simulated in a laboratory and the second will be to identify superior genotypes for larval survival in order to help alleviate the current problems faced by hatcheries with poor larval performance.

Harbour Branch Florida Atlantic University. John Scarpa reported on the Molluscan Research Program in Florida Atlantic University. Their apple snail breeding program is in its fourth generation and is thriving. There is also work being done on conch reproductive stock enhancement. In bivalves the sunray Venus clam can now be cultured using methods developed for hard clam culture. The next step is to identify methods of brood stock handling and spawning, establish hatchery methods, evaluate growth and survival during grow out, test market at wholesalers. With Mercenaria mercenaria there continues to be increased clam mortality in SE Florida. Hybrids have been created and their performance is currently being tested in the field. So far, hybrids show superior growth. They are also looking at larval dispersal of C. virginica along Eastern Florida

Industry Participation: Taylor Shellfish Farms. Joth Davis gave a first hand view of the problems being faced by the oyster industry in the Pacific Northwest. He then talked about the line evaluation and development process at Taylor Shellfish. Taylor run a 4-phase rearing schedule to evaluate hybrid lines and measure additive and non-additive contribution to yield. Larval rearing systems have been changed to low volume, high density flow through system. This increased ability to test more families (50 to 100 lines per cohort), they aim to optimize seed testing methods to minimize time and reduce cost. They found that cage culture produces poor results and rotating seed bag worked better; cylinder worked best. They are also working on Manila clam breeding. There is now a need to develop molecular tools for genetic typing and pedigree analysis

Invited speaker Dr. Peter Kube CSIRO, Australia: Peter Kube gave a presentation titled Genetic Improvement of the Pacific Oyster- A quantitative genetics approach down under. He gave an overview of the oyster industry in Australia. Two species are farmed-the Pacific and Sydney rock oysters. The industry generates approximately $50M annually. 20% of the production is from genetically improved seed. The breeding program began in 1997 and to date there has been 10 year classes with 24-60 families per year class but because the breeding objective was defined by farmers preferences progress was slow and not cumulative. A new direction was taken based on an economic model and the economic values of traits were used to identify the breeding goals. He described an economic model based on economic weights. The economic weight of a trait is the dollar value of a change in the trait and it can be expressed as change in production cost. He then went on to describe the ways in which quantitative genetics was used to select potential broodstock. Data is combined from families across all year classes and analyzed using ASREML, a statistics program that allows multivariate models while at the same time combining information on all traits and all animals in a pedigree (up to 14,000 individuals). This program is used to generate EBVs (estimated breeding values) for all traits between families, the higher the EBV the better the family. The trait EBVs are then combined into a single index using economic weights and families are selected for breeding based on this index. Future work on the program included setting up a database to streamline the process, understand the genetics of survival and increase the number of families tested each year.

Opportunities for collaborative research Gary Jensen USDA CSREES, and Jeff Silverstein USDA. Gary Jensen and Jeff Silverstein gave a joint presentation on ways in which grant applications can be strengthened by incorporating various disciplines and including outreach activities. The key is to keep stakeholders in view and talk to program managers before applying. USDA and CSREES priority areas were identified.

Accomplishments

WERA-099 provided a forum for US and international molluscan geneticists, physiologists and pathologists to exchange ideas and information on quantitative and molecular genetics, reproduction, diseases, chromosome and genetic manipulation techniques, broodstock management and breeding programs. Highlights of the 2008 meeting included: A one day workshop entitled 'From Falconer to the Field- the practical application of quantitative genetics theory to existing oyster breeding programmes' & 'Gene maps to field maps- a synergy of molecular and quantitative tools targeted at improved oyster performance'. <br /> This workshop:<br /> 1) increased the awareness of US scientists about the need to design their breeding schemes with additive genetic variance in view and has resulted in a reassessment of our breeding program designs; 2) highlighted the areas of collaborative research as this is essential to tackle the disease problems faced by the aquaculture industries across the US; 3) intensified efforts towards developing multidisciplinary projects; 4) increased knowledge about the relative performance of selectively bred strains, hybrids, and triploids in farms in the Eastern US; 5) intensified efforts towards marker development; 6) pushed for development of molecular diagnostic tools and laboratory disease challenges for diseases affecting the bivalve industry; 7) highlighted efforts towards the identification of molecules involved in bivalve immune responses and resistance to stress; and 8) demonstrated progress towards mapping and genome sequencing. <br /> Multi-disciplinary and inter-regional exchanges among participants revealed new directions of scientific enquiry and facilitated formulation of new research questions, particularly in quantitative genetics. The meeting also provided a forum to stimulate and develop collaborations between US and Australia. <br />

Publications

Batista FM, Arzul I, Pepin JF, Ruano F, Friedman CS, Boudry P, Renault T (2007) Detection of ostreid herpesvirus 1 DNA by PCR in bivalve molluscs: a critical review. J Virol.Methods 139:1-11<br /> <br /> Burge CA, Judah LR, Conquest LL, Griffin FJ, Cheney DP, Suhrbier A, Vadopalas B, Olev PG, Renault T, Friedman CS (2007) Summer seed mortality of the Pacific oyster, Crassostrea gigas thunberg grown in Tomales Bay, California, USA: The influence of oyster stock, planting time, pathogens, and environmental stressors. J.Shellfish Res 26(1):163-172<br /> <br /> Camara MD, Evans S, Langdon C (2007) Inbreeding effects on growth and survival in a naturalized population of the Pacific oyster (Crassostrea gigas) revealed using molecular marker-based estimates of parental relatedness. Aquaculture 272:S247<br /> <br /> Curole JP, Hedgecock D (2007) Bivalve Genomics: Complications, Challenges, and Future Perspectives. Aquaculture Genome Technologies. Jul 2007, pp 525-543.<br /> <br /> Gomez-Leon J, Villamill L, Salger SA, Sallum RH, Remacha-Trivino A, Leavitt DF, Gomez-Chiarri M (2008) Survival of eastern oysters Crassostrea virginica from three lines following experimental challenge with bacterial pathogens. Dis Aquat Organ 79:95-105<br /> <br /> Jenny MJ, Chapman RW, Mancia A, Chen YA, McKillen DJ, Trent H, Lang P, Escoubas JM, Bachere E, Boulo V, Liu ZJ, Gross PS, Cunningham C, Cupit PM, Tanguy A, Guo X, Moraga D, Boutet I, Huvet A, De GS, Almeida JS, Warr GW (2007) A cDNA microarray for Crassostrea virginica and C. gigas. Mar Biotechnol.(NY) 9:577-591<br /> <br /> Moss JA, Burreson EM, Cordes JF, Dungan CF, Brown GD, Wang A, Wu X, Reece KS (2007) Pathogens in Crassostrea ariakensis and other Asian oyster species: implications for non-native oyster introduction to Chesapeake Bay. Dis Aquat Organ 77:207-223<br /> <br /> Moss JA, Xiao J, Dungan CF, Reece KS (2008) Description of Perkinsus beihaiensis n. sp., a new Perkinsus sp. parasite in oysters of Southern China. J Eukaryot.Microbiol 55:117-130<br /> <br /> Quilang J, Wang S, Li P, Abernathy J, Peatman E, Wang Y, Wang L, Shi Y, Wallace R, Guo X, Liu Z (2007) Generation and analysis of ESTs from the eastern oyster, Crassostrea virginica Gmelin and identification of microsatellite and SNP markers. BMC Genomics 8:157<br /> <br /> Roberts S, Gueguen Y, de LJ, Goetz F (2008) Rapid accumulation of an interleukin 17 homolog transcript in Crassostrea gigas hemocytes following bacterial exposure. Dev Comp Immunol 32:1099-1104<br /> <br /> Taris N, Lang RP, Camara MD (2008) Sequence polymorphism can produce serious artefacts in real-time PCR assays: hard lessons from Pacific oysters. BMC Genomics 9:234<br /> <br /> Wang Y, Guo X (2007) Development and characterization of EST-SSR markers in the eastern oyster Crassostrea virginica. Mar Biotechnol.(NY) 9:500-511<br />

Impact Statements

1. At the 2009 meeting it became obvious that funding required to conduct fundamental breeding experiments that generate parameters and information needed to optimize breeding programs in severely lacking.
2. Discussions between researchers at the workshop showed that work is needed to modify current breeding programs in order to make them more efficient.
3. Conversations between researchers should result in the development of markers for marker-assisted selection that could be used in more efficient breeding programs to produce disease-resistant strains. Future efforts will be geared to marked validation.
4. Research efforts are geared to provide industry with stocks, families, and lines that show improved performance.
5. International efforts to map the genome of the Pacific and Eastern oyster and to sequence the genome of the Pacific oyster are moving forward. Exchange of information on the development of new maps and mapping techniques will lead to a better understanding of the genetic basis of disease resistance in Eastern oysters as well as growth and survival in farmed Pacific oysters.

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Report Information

Participants

Meeting minutes includes list of those present.
URL:
https://catalysttools.washington.edu/workspace/sr320/8608/41770

Brief Summary of Minutes

Accomplishments

" Increased level of awareness among participants of the problems facing West coast shellfish and hatcheries in dealing with the effects of coastal upwelling and acidification of coastal waters. <br /> <br /> " Learned from international participants about strategies to transfer selected broodstock and information to industry <br /> <br /> " New collaborative projects have been developed to address the impacts of ocean acidification on West coast shellfish<br /> <br /> " Dr. Reece served on the Oyster Restoration Workshop Panel for the Hudson River Foundation, Aug. 11-12, 2009, NYC, New York. <br /> <br /> " During this past year the Reece lab continued development and implementation of genetic markers for the hard clam Mercenaria mercenaria<br />

Publications

Publications for the current year only.<br /> <br /> <br /> Batista FM, Boudry P, Dos Santos A, Renault T, Ruano F (2009). Infestation of the cupped oysters Crassostrea angulata, C. gigas and their first generation hybrids by the copepod Myicola ostreae: differences in susceptibility and host response. Parasitologia 136 (5): 537-543.<br /> <br /> Sauvage C, Pépin JF, Lapègue S, Boudry P, Renault T (2009). Ostreid herpes virus infection in families of the Pacific oyster, Crassostrea gigas, during a summer mortality outbreak: differences in viral DNA detection and quantification using real-time PCR. Virus Research, 142: 181187<br /> <br /> Sauvage C, Boudry P, Lapègue S. Identification and characterization of 18 novel polymorphic microsatellite makers derived from expressed sequence tags in the Pacific oyster Crassostrea gigas. Molecular Ecology Notes, 9(3): 853-855.<br /> <br /> Lallias D, StockdaleR, Boudry P, Beaumont AR, Lapègue S. Characterization of 27 microsatellite loci in the European 1 flat oyster Ostrea edulis. Molecular Ecology Resources, 9(3): 960-963.<br /> <br /> Lallias D, Gomez-Raya L, Haley CS, Arzul I, Heurtebise S, Beaumont AR, Boudry P, Lapègue S (2009). Combining two-stage testing and interval mapping strategies to detect QTL for resistance to bonamiosis in the European flat oyster Ostrea edulis. Marine Biotechnology 11:570584.<br /> <br /> Lallias D, Stockdale R, Boudry P, Lapègue S, Beaumont A (2009). Characterization of ten microsatellite loci in the blue mussel Mytilus edulis. Journal of Shellfish Research 28(3): 547-551.<br /> <br /> Fleury E, Huvet A, Lelong C, de Lorgeril J, Boulo V, Gueguen Y, Bachère E, Tanguy A, Moraga D, Fabioux C, Lindeque P, Shaw J, Reinhardt R, Prunet P, Davey G, Lapègue S, Sauvage C, Corporeau C, Moal J, Gavory F, Wincker P, Moreews F, Klopp C, Mathieu M, Boudry P, Favrel P (2009). Generation and analysis of a 29,745 unique Expressed Sequence Tags from the Pacific oyster (Crassostrea gigas) assembled into a publicly accessible database: the GigasDatabase. BMC Genomics, 10: 341.<br /> <br /> Arias A, Freire R, Boudry P, Heurtebise S, Méndez J, Insua A (2009). Single nucleotide polymorphism for population studies in the scallops Aequipecten opercularis and Mimachlamys varia. Conservation Genetics, 10:14911495.<br /> <br /> Cannuel R, Beninger PG, McCombie H, Boudry P (2009). Gill development, functional and evolutionary implications in the blue mussel Mytilus edulis (Bivalvia: Mytilidae). Biological Bulletin 217: 173-188.<br /> <br /> Normand J, Ernande B, Haure J, McCombie H, Boudry P (2009). Reproductive effort in Crassostrea gigas: comparison of 5-month-old diploid and triploid oysters issued from natural crosses or chemical induction. Aquatic Biology 7(3): 229-241<br /> Taris N, Boudry P, Bonhomme F, Camara MD, Lapègue S (2009). Mitochondrial and nuclear DNA analysis of genetic heterogeneity in temporal recruitment cohorts of the European flat oyster, Ostrea edulis. The Biological Bulletin 217(3): 223-241 <br /> <br /> Dégremont L, Bédier E, Boudry P (2010). Summer mortality of hatchery-produced Pacific oyster spat (Crassostrea gigas). II. Response to selection for survival and its influence on growth and yield. Aquaculture 299: 21-29.<br /> <br /> Dégremont L, Boudry P, Ropert M, Samain JF, Bédier E, Soletchnik P (2010). Effects of age and environment on survival of summer mortality by two selected lines of the Pacific oyster Crassostrea gigas. Aquaculture 299: 44-50. <br /> <br /> Fleury E, Moal J, Boulo V, Daniel J-Y, Mazurais D, Hénaut A, Boudry P, Favrel P, Huvet A. Identification of new genes associated with summer mortality in the oyster Crassostrea gigas. Marine Biotechnology, in press.<br /> <br /> Pernet F, Barret J, Marty C, Moal J, Le Gall P, Boudry P. Environmental anomalies, energetic reserves and fatty acid modifications in oysters coincide with an exceptional mortality event. Marine Ecology Progress Series, in press.<br /> <br /> Sauvage C, Boudry P, de Koning DJ, Haley CS, Heurtebise S, Lapègue S. Quantitative Trait Loci for resistance to summer mortality and OsHV1 load in the Pacific oyster (Crassostrea gigas). Animal Genetics, in press. <br /> <br /> Huvet A, Normand J, Fleury E, Quillien V, Fabioux C, Boudry P. Reproductive effort of Pacific oysters: a trait associated with susceptibility to summer mortality. Aquaculture, in press.<br /> <br /> Geay F, Santigosa I Culi E, Corporeau C, Boudry P, Dreano Y, Corcos L, 4, Bodin N, Vandeputte M, 7, Zambonino-Infante JL,Mazurais D, Cahu CL. Regulation of FADS2 expression and activity in European sea bass (Dicentrarchus labrax, L.) fed a vegetable diet. Comparative Biochemistry and Physiology, in press.<br /> <br /> Lallias D, Boudry P, Lapègue S, King JW, Beaumont AR. Strategies for the retention of high genetic variability in European flat oyster (Ostrea edulis) restoration programmes. Conservation Genetics, in press.<br /> <br /> Lang, P.L., Bayne, C.J, Camara, M.D., Cunningham, C., Jenny, M.J., and Langdon C.J. 2009. Transcriptome profiling of selectively-bred Pacific oyster Crassostrea gigas families that differ in tolerance of heat shock. Marine Biotechnology, 11: 650-668<br /> <br /> Lang, R.P., Langdon, C.J., Taris, N.G. and M.D. Camara. Can laboratory assays be used to predict field performance of Pacific oyster (Crassostrea gigas) families planted in coastal waters? Aquaculture, accepted.<br /> <br /> Costa de Melo, A.G., Gardunho, D.C.L., Schneider, H., Sampaio, I., Gaffney, P.M., Reece, K.S., Beasley, C.R. and C.H. Tagliaro. (in press) Molecular identification, phylogeny and geographic distribution of Brazilian mangrove oysters (Crassostrea). Gen. Mol. Biol.<br /> <br /> Xiao, J. Cordes, J.F., Wang, H., Guo, X., and K.S. Reece. (in press) Population genetics of Crassostrea ariakensis in Asia inferred from microsatellite markers. Mar. Biol.<br /> <br /> Roberts SB, Goetz G, White S, Goetz F (2009) Analysis of genes isolated from plated hemocytes of the Pacific Oyster, Crassostrea gigas. Marine Biotechnology. 11:24-44<br /> <br /> DeFaveri J, Smolowitz R, Roberts, S. (2009) Development and validation of a real-time quantitative PCR assay for the detection and quantification of Perkinsus marinus in the Eastern oyster, Crassostrea virginica. Journal of Shellfish Research<br /> <br /> Books and books chapters<br /> <br /> Boudry P (2009) Genetic variation and selective breeding in hatchery-propagated molluscan shellfish. In Burnell G & Geoff A (eds): New technologies in aquaculture: Improving production efficiency, quality and environmental management. Woodhead Publishing Limited, Cambridge, U.K, pp. 87-108.<br /> <br /> Beaumont A, Boudry P, Hoare K (2010). Biotechnology and genetics in Fisheries and Aquaculture (2nd ed.). Wiley-Blackwell, Chichester, UK. 202 p.<br />

Impact Statements

1. Establishment of a database dedicated to C. gigas EST (Fleury et al., BMC Genomics 2009, 10: 341).
2. First use of RNA interference in C. gigas, demonstrating the role of vasa in germ cell development (Fabioux et al., FEBS journal 276: 2566-2573)
3. QTL for resistance to: - summer mortality and OsHV-1 load in C. gigas (Sauvage et al., Animal Genetics, in press) - bonamiosis in the European flat oyster O. edulis (Lallias et al., Marine Biotechnology 11: 570584).
4. Genetic identification of oyster samples found lose in the Coan River. Oysters confirmed to be triploid C. ariakensis. Report sent to VIMS Dean/Director, VIMS DRAS and VMRC.
5. Genetic identification of oysters found on crab pots in the York River. Oysters confirmed to be C. virginica and not C. ariakensis as suspected. March 2009. Report sent to VIMS Dean/Director, VIMS DRAS and VMRC.

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