Annual/Termination Reports:

[05/17/2002] [06/11/2003] [04/05/2004] [04/19/2005] [05/25/2006]

Report Information

Participants

Allen, Stan, ska@vims.edu Virginia Institute of Marine Sciences;
Boudry, Pierre, pierre.boudry@ifremer.fr, IFREMER, La Tremblade;
Brake, John, john.brake@hmsc.orst.edu, Hatfield Marine Science Center, Oregon State University; Camara, Mark, camara@vims.edu, Virginia Institute of Marine Sciences; Davis, Joth, jdavis32@mindspring.com, Taylor Shellfish Farms; Gaffney, Patrick, pgaffney@udel.edu, University of Delaware; Guo, Ximing, xguo@hsrl.rutgers.edu, Rutgers University; Samain, Jean-Francois, Jean.Francois.Samain@ifremer.fr, IFREMER, Brest; Hedgecock, Dennis, dehedgecock@ucdavis.edu; Bodega Marine Lab, University of California- Davis; Langdon, Chris, chris.langdon@hmsc.orst.edu, Hatfield Marine Sciences Center, Oregon State University; Li, Li, lili@hsrl.rutgers.edu, Rutgers University; Matson, Sean, sean.matson@hmsc.orst.edu, Hatfield Marine Sciences Center, Oregon State University; Meritt, Don, merit@hpl.umces.edu, Horn Point Laboratory, University of Maryland; Nicola, Jean Louis, jlnicola@ifremer.fr, IFREMER; Reece, Kimberly, kreece@vims.edu, Virginia Institute of Marine Sciences; Wikfors, Gary, gary.wikfors@noaa.gov, NOAA; Wilbur, Amy, wilbura@uncwil.edu,CMS UNCW

Brief Summary of Minutes

The meeting started at 9 AM with an introduction by Stan Allen who described the history of WCC-99 & its purpose.



Pierre Boudry reported on research of the genetics & pathology program at the IFREMER station at La Tremablade. Primary areas of research in genetics of the European & Pacific oyster are: molecular taxonomy, population genetics, phenotypic evaluation, cytogenetics & improvement of stocks by genetic selection.



Species-specific arkers have been developed for various oyster species & varieties collected from around the world. Based on their data, the mangrove oyster Crassostrea gasa is found in both Africa & South America but Crassostrea rhizophora is only found in South America. The population genetics of the European oyster has been studied & the distribution of different stocks described on the Mediterranean & Atlantic coasts. In addition, the ecophysiologies of Crassostrea gigas & Crassostrea angulata have been compared to show that the specific growth rate of C. angulata is lower than that of C. gigas. Cytogenetic studies have shown that there is correlation between aneuploidy & growth of Pacific oysters. Genomic tags are being developed for each chromosome. Some pairs of chromosomes appear to be lost more commonly than others. Pollution results in increased rates of chromosome loss. Genetic selection has resulted in improved survival of European oysters exposed to Bonamia. A large inter-institution project called MOREST has begun in France to study the heritability of growth & survival of Pacific oysters.



Jean-Francois Samain described the MOREST program, The five-year program is multidisciplinary involving 15 French laboratories & 22 teams. The five main tasks are 1) to prepare tools in all disciplines to study summer mortality, 2) to characterize the environment & identify possible causal factors, 3) to examine interactions among possible causes, including both environmental & physiological factors, 4) to identify potential pathogens using a national network of laboratories, 5) to develop long-term monitoring programs for oyster growing areas susceptible to summer mortality. Preliminary genetic selection indicated a strong genetic effect on summer mortality with family survival ranging from 17 to 97%. It was noted that mortalities do not occur until seawater temperatures rise to >19 oC & they often occur shortly before spawning.



Gary Wikfors discussed the importance of understanding feeding & nutrition of mollusks as part of the domestication process. He described an apparatus to vary the ration & frequency of feeding shellfish on algal diets. Rations of 5% whole body weight per day were optimal for growth but food conversion efficiencies were best at 2% ration per day.



Pat Gaffney described the use of various genetic markers to distinguish different populations of the Eastern oyster on the East & Gulf coasts, US. Use of 16S mitochondrial DNA sequences was useful in distinguishing Gulf from East Coast populations, with over 33 polymorphic restriction sites identified.



Pat described his work on sequencing nuclear DNA of the Eastern oyster. About 1446 sequences have been described & there is evidence of a non-autonomous DNA transposon (CvA). He also reported that only 50% of the flanking regions of microsatellites of the Eastern oyster are unique, resulting in possible confusion in amplification of microsatellite markers.



Ximing Guo Numerous genetic projects are being undertaken, including 1) breeding & evaluation of NE stocks of Eastern oysters, 2) aneuploidy & triploidy in oysters, 3) production of triploid & tetraploid Eastern oysters & hard clams, 4) genetic expression analysis of oyster responses to Dermo & MSX infections, 5) gene mapping of Eastern & Pacific oysters, 6) physical mapping of chromosomes (FISH).



About 42,000 tetraploid Eastern oysters have been successfully produced. High mortality of the larvae & spat meant that large numbers of broodstock were needed to produce sufficient larvae. Tetraploid Eastern oysters were not stable & 16% showed "mosaic" ploidy after three months, a rate that was greater than for triploids.



An AFLP map has been produced for the Eastern oyster with 170 markers identified. A second map has been produced for the Pacific oyster with 145 markers.



Suppression subtraction libraries of expressed genes for Eastern & Pacific oysters exposed to Dermo & MSX infection have been produced. A wide range of gene expression responses have been detected & are currently being analyzed.



Ami Wilbur described her work on comparing different oyster stocks planted at various sites on the East coast. Stocks were identified using two 16S mtDNA loci. Four % of Eastern oyster seed sampled from "wild" oyster beds likely originated as seed produced from planted Louisiana oysters at a site in North Carolina.



Ami reported on work to determine the effectiveness of enhancement programs for Bay scallops. Research in Florida indicated that three years of scallop enhancement had no detectable effect in contributing to the recovery of scallop populations in that state. However, in Chincoteague Bay, efforts by Virginia agencies were more effective. The closed nature of the bay may have contributed to the positive results of the enhancement program.



Don Meritt described plans for a new shellfish hatchery at the Horn Point Laboratories, University of Maryland, which is planned for completion next spring. The hatchery will cost an estimated $25 million & will receive a flow-through seawater supply. The hatchery will also have a quarantine lab that will allow importation of non-native species.



Stan Allen reported on the results of the Williamsberg conference held in October, 2001, at which the aquaculture potential of triploid Suminoe oysters Crassostrea ariakensis was discussed. He recounted the history of the study of non-native species at VIMS leading up to the current industry trials with about 60,000 out-planted triploid oysters. Growth & survival results indicate that the performance of Suminoe oysters at test sites is much better than that of Eastern oysters.



A conclusion of the science panel at the Williamsburg Conference was that triploid aquaculture of Suminoe oysters would inevitably lead to the introduction of diploids either through illegal introductions, ploidy reversions, lapses in biosecurity or harvest loss. Any scenario for triploid introductions should include consideration of the fate of diploid introductions & possible effects of competition with Eastern oysters, return of the Chesapeake to a benthic-dominated system & fouling of yachts, marine structures & intake pipes. The conference recommended that more work should be undertaken to study the ecological consequences of diploid introductions. In June 2002, a committee of the National Academy of Sciences will undertake a review of the issue.



Mark Camara discussed some concepts of breeding programs. He noted that one needs to develop fast-growing lines for aquaculture using high-tech procedures, while for restoration purposes one can adopt a low-tech approach. The pros & cons of adopting a "walk-back" approach to genetic selection were discussed.



Kim Reese described research that focused on developing markers to discriminate between hatchery & natural stocks as well the identification of genes useful in selecting for disease resistance, growth rate & final size at harvest. Her lab is focusing on developing microsatellite markers using primers that amplify only at a single locus. She reported a high incidence of null alleles for many microsatellites but found that dinucleotide microsatellites were associated with fewer null alleles that tetranucleotide microsatellites. Using RFLP analysis, Kim reported that the DERBY line of disease-resistant Eastern oysters was highly inbred and, therefore, has limited use in restoration. CROSBreed lines were less inbred & showed more promise for use in restocking.



Dennis Hedgecock described the approach of Lynx Therapeutics for measuring genetic expression of oysters using massively parallel analysis. The method is based on the use of millions of microbeads each providing a unique 20 base pair signature sequence. A total of about 3 million sequences can be analyzed at a time. Dennis used the method to compare expressed genes in inbred & outbred lines (MBP families 1.35 & 1.51). About 2 to 3% of the expressed genes were non-additive (i.e. their expression was either greater or less than the average of the two parental inbred lines). Of these non-additive genes, about one third were over-dominant & one third were under-dominant. A total of 120 were significantly non-additive in both hybrids & are candidate genes of interest in explaining hybrid vigor in these lines. The 20 base-pair sequences will be used to produce longer 250 base-pair sequences for GenBank searches. The non-additively expressed genes of interest will be evaluated as QTLs for traits of interest.



Dennis also described progress in mapping the genome of Pacific oysters. Optimal PCR methods have been developed for 190 microsatellite primers & 79 markers have been mapped. A high proportion (49/96) of microsatellites are associated with null alleles indicting a high level (0.0123) of polymorphism - a rate that is ten times that reported for humans. Dennis estimated that this level of polymorphism in oysters would result in a SNP every 80 base pairs.



Joth Davis reported on the results of the WRAC project for cross-breeding inbred lines of Pacific oysters. Taylor Farms Inc. has established a bivalve research & testing center with a capacity for 50 x 100 liter tanks supplied with 0.2 micron-filtered, UV-treated seawater. The nursery has 96 upwellers for growth of juveniles. The grow-out system consists of bag suspended from long-lines. Large differences in the growth of seed from various hybrid lines were observed.



Chris Langdon described progress for the Molluscan Broodstock Program (MBP). Analysis of yields of Pacific oysters derived from selected broodstock showed a realized heritability of 9.4% for improvement in yield compared with yields of non-selected "wild" oysters. Both growth & survival affect yield, with the relative proportion differing among sites & cohorts. In Tomales Bay, 79% of the variation in yield among families was due to differences in survival.



John Brake discussed methods to describe shell & mantle color as well as cup shape of Pacific oysters. These parameters will be added to descriptions of the various MBP families on the MBP web site. Future experiments are planned to determine the heritability of shell & mantle color as well as shell shape.



Sean Matson described the results of a commercial-scale test of MBP seed. The performance of seed derived from crossing MBP broodstock was compared with that of seed from a standard commercial cross. The seed were produced on a commercial-scale at Taylor‘s hatchery & planted either on-cultch oysters in Samish Bay or as singles in bags in Totten Inlet. Microsatellite analysis of the seed at planting indicated a contamination level of about 8%. The parentage of the contaminants was unknown, indicating that contamination likely occurred in the hatchery or nursery.



At harvest, MBP-derived oysters were heavier with a lower percentage of extra smalls & debris (empty shells) compared with controls. In support of this finding, the volume ratio of harvested oysters to discard material was 5 to 1 for MBP oysters but only 2:1 for control oysters. There was a likely mix-up of oyster meats during shucking as meat weight data for the MBP & control oysters were not correlated with whole weights - a very unlikely result.



Stan Allen volunteered to remain Chairperson for a second year with Chris Langdon as Secretary. The venue for the next meeting was not determined. Adjourned at 5.30 pm.

Accomplishments

Principal accomplishments of this group include major work on the West Coast in tetraploidy research, primarily coming out of the MBP & USDA Western Regional Aquaculture Consortium funded projects. Triploid & tetraploid technology has moved from research to application, supported, in part, by USDA-SBIR funding. MBP select stocks are being propagated commercially. The WRAC project has one commercial test of hybrid oyster yield in the field; however, more are planned. There is vigorous industry support for these activities.<br /> <br><br /> <br>Work parallel to that of the West Coast concerns the Eastern Oyster & includes investigations of aneuploidy, triploidy & tetraploidy & some important disease investigations related to Dermo & MSX. FISH is being used to map the chromosomes of the Eastern Oyster.<br /> <br><br /> <br>There have been important key papers published relating to the causes & uses of hybrid vigor & the development of microsatellite DNA markers for confirming parentage in experiments & for gene-mapping. A number of the members of WCC-099 also participate in the Regional project NE-186, Genetic Maps of Aquaculture Species. A genetic linkage map of the Pacific Oyster based on microsatellite technology has been developed & there has been analysis of gene expression in hybrid Pacific Oysters by massively parallel signature sequencing.<br /> <br><br /> <br>There have been important research efforts from members of this group to identify & conserve the Kumamoto oyster. Work has also taken place on the nutrition of mollusks in relationship to the domestication process.

Publications

Launey, Sophie; Hedgecock, Dennis. High genetic load in the Pacific oyster Crassostrea gigas. 2001. Genetics 159: 255-265.<br /> <br><br /> <br>Hallerman, E., M. Leffler, S. Mills, and S.K. Allen, Jr. 2002. Aquaculture of triploid Crassostrea ariakensis in Chesapeake Bay: A Symposium Report. Md/Va Sea Grant Publication, UM-sg-ts-2002-01/VSG-02-03, 20 pp.<br /> <br><br /> <br>Sophie Hubert, Gang Li, Dennis Hedgecock. 2002. Development of a genetic linkage map, using microsatellite markers in the Pacific oyster, Crassostrea gigas. http://www.intl-pag.org/pag/10/abstracts/PAGX_W16.html<br /> <br><br /> <br>Hedgecock, D., Lin, J.-Z. DeCola, S., Haudenschild, C., Meyer, E., Manahan D.T., Bowen, B. 2002. Analysis of gene expression in hybrid Pacific oysters by massively parallel signature sequencing.<br /> <br>http://www.intl-pag.org/pag/10/abstracts/PAGX_W15.html

Impact Statements

1. Triploid & tetraploid technology has moved rapidly from research to application supported by various funding sources, e.g., USDA-SBIR, to members of this project. MBP select stocks are being reared on farms & are being propagated commercially by two west coast firms. West Coast industry is contributing the essential facilities & manpower to support the projects. Their support is a clear indication of their confidence in USDA-supported research for hybrid vigor & a commitment to crossbreeding.

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Report Information

Participants

Ristow,Sandra(ristow@wsu.edu)-Washington State University;
Rawson,Paul(prawson@maine.edu)-Rutgers University;
Guo,Ximing(xguo@hsrl.rutgers.edu)-Rutgers University;
Fransois,Samain Jean-IFREMER Center of Brest;
Jeanne,Moal-IFREMER Center of Brest;
Camara,Mark(camara@vims.edu)-VIMS;
Robinson,Anja-Oregon State University;
Davis,Joth(jdavis32@mindspring.com)-Taylor Shellfish Farm;
Langdon,Chris(chris.langdon@oregonstate.edu)-Oregon State University;
Reeve,Kim(kreece@vims.edu)-VIMS;
Wilbur,Ami E(wilbura@uncw.edu)-UNCW;
Janke,Achim(achim@cawthron.org)-Cawthoron Institute.

Brief Summary of Minutes

Accomplishments

Ximing Guo Director at the Haskin Shellfish Research Laboratory, Rutgers University conducts research in three areas of molluscan genetics: selective breeding, ploidy manipulation and genomics. Guo&lsquo;s team has continued the long-term breeding program that was first established by the late Dr. Haskin in early 1960s. Recent studies have shown that the hybrid cross between the Rutgers disease-resistant strain and the Frank M. Flower&lsquo;s fast-growing variety provides the highest yield by combining disease-resistance and superior growth. In ploidy manipulation, his laboratory produced breeding populations of tetraploid and disease-resistant eastern oysters that are being used for triploid production by the oyster culture industry. Triploid oysters grow significantly faster than diploids. In genomics, Guo&lsquo;s team published a moderately-dense genetic linkage map for the eastern oyster, providing a first step toward the mapping of disease-resistant genes.<br /> <br><br /> <br>Dennis Hedgecock<br /> <br><br /> <br>A linkage map for the Pacific oyster Crassostrea gigas,which would be an invaluable tool for improving commercial stocks, has been developed using 100 microsatellite markers. Thus far, ten linkage groups have been found, a number that corresponds to the haploid number of chromosomes. The male consensus map has 89 loci and covers about 700 map units (centimorgans, cM); the female map has 86 loci and covers about 750 cM, values in good agreement with cytological observations. Significant differences have been detected in recombination rates and gene orders among families and even among siblings, which suggests polymorphism for chromosomal rearrangements exists in natural populations. The linkage map has two immediate uses, to map the location of previously detected recessive deleterious mutations in F3 and F4 populations and to map regions of the genome that contribute to variation in quantitative and economically important traits; research in both areas is in progress. The markers and maps are shared with WCC-99, providing common tools for use by members of this group.<br /> <br><br /> <br>Paul Rawson<br /> <br><br /> <br>Our laboratory is just beginning to re-establish a program for genetic improvement with the eastern oyster, Crassostrea virginica, in Maine. Although, most of the genetic improvement programs for this species have been concerned with disease-resistance, our primary concern is to develop stocks with improved cold-water growth rates. Even so, WCC-99 has proven to be invaluable to our efforts. The enhanced communication among colleagues working on oyster genetics that is engendered through WCC-99, particularly at the annual meetings held in conjunction with the National Shellfisheries Conference, has helped us to design our broodstock development program in a way that complements other on-going programs, avoid needless overlap in effort, and to develop stronger collaborations.<br /> <br><br /> <br>Christopher Langdon<br /> <br><br /> <br>1) Exchange of technical information<br /> <br>2) Advice for improvement of the USDA-supported Molluscan Broodstock Program<br /> <br>3) Design of commercial trials with selected and hybrid oyster families<br /> <br>4) Co-ordination of research efforts for multi-PI projects<br /> <br>5) Establishment of stronger links with French colleagues working on summer<br /> <br> mortality of Pacific oysters.

Publications

Yu, Z. and X. Guo. 2003. Genetic linkage map of the eastern oyster<br /> <br>Crassostrea virginica Gmelin. Biol. Bull. 204: 327-338.<br /> <br><br /> <br>Peruzzi, S. and X. Guo. 2002. Tetraploid induction by meiosis inhibition<br /> <br>with cytochalasin B in the dwarf surfclam, Mulinia lateralis Say: effects of<br /> <br>temperature. J. Shellfish Research, 21(2): 677-684.<br /> <br><br /> <br>Wang, Z., X Guo, S.K. Allen, Jr. and R. Wang. 2002. Heterozygosity and body<br /> <br>size in triploid Pacific oysters, Crassostrea gigas Thunberg, produced from<br /> <br>meiosis II inhibition and tetraploids. Aquaculture, 204(3-4):337-248.<br /> <br><br /> <br>Wang, Y., Z. Xu and X. Guo. 2001. A centromeric satellite sequence in the<br /> <br>Pacific oyster, Crassostrea gigas (Thunberg) identified by fluorescence in<br /> <br>situ hybridization. Marine Biotechnology, 3: 486-492.<br /> <br><br /> <br>Yang, H., T. Gallivan, X. Guo and S.K. Allen, Jr. 2000. A method for preserving oyster tissue samples for flow cytometry. J. Shellfish Research, 19(2):835-839.<br /> <br><br /> <br>Bucklin, K. A. 2002. Genetic Load in Two Generations of the Pacific Oyster Crassostrea gigas. Ph.D. dissertation, genetics, University of California, Davis.<br /> <br><br /> <br>Hedgecock, D., S. Hubert, G. Li and K. Bucklin. 2002. A genetic linkage map of 100 microsatellite markers for the Pacific oyster Crassostrea gigas. Journal of Shellfish Research 21:381. <br /> <br><br /> <br>Hubert, S., B. J. Landau, L. English, X. Guo, and D. Hedgecock. 2002. Genetics and linkage groups of microsatellite markers in the Pacific oyster, Crassostrea gigas using trisomics. Aquaculture 204:216. http://www.elsevier.com/locate/aquaculture <br /> <br><br /> <br>Li, G., S. Hubert, K. Bucklin, V. Ribes and D. Hedgecock. 2002. Characterization of 79 microsatellite DNA markers in the Pacific oyster Crassostrea gigas. Molecular Ecology Notes 3:228-232.<br /> <br><br /> <br>Hedgecock, D., S. Hubert and K. A. Bucklin. 2002. Linkage and gene-centromere maps of the Pacific oyster Crassostrea gigas. http://www.intl-pag.org/pag/11/abstracts/W05_W36_XI.html<br /> <br><br /> <br>Langdon, C.J., Evans, F., Jacobson, D. and Blouin, M. Yields of cultured<br /> <br>Pacific oysters Crassostrea gigas Thunberg improved after one generation of<br /> <br>selection. Aquaculture, 220: 227-244.<br /> <br><br /> <br>Brake, J., Evans, F. and C. Langdon. 2002. Evidence for genetic control of<br /> <br>pigmentation of shell and mantle edge in selected families of Pacific<br /> <br>oysters, Crasssotrea gigas. Aquaculture.

Impact Statements

1. A moderately dense genetic linkage map has been produced for the Eastern Oyster which provides a first step toward the map of disease-resistance genes. This will allow the mapping of resistance to MSX and Dermo.
2. A linkage map for the Pacific Oyster has been produced utilizing 100 microsatellite markers and ten linkage groups have been found. The map is useful for locating recessive deleterious mutations in the F3 and F4 generations.

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Report Information

Participants

Ristow,Sandra(Ristow@wsu.edu)-Washington State University;
Reece,Kimberly(kreece@vims.edu)-Virginia Institute of Marine Science;
Camara,Mark(Mark.Camara@oregonstate.edu)-USDA-ARS;
Evans,Ford(ford.evans@oregonstate.edu)-Oregon State University;
King,Nick(Nick.King@cawthron.org.nz)-Cawthron;
Dewey,Bill(billd@taylorshellfish.com)-Taylor Shellfish Company;
Cudd,Sue(suecudd@aol.com)-Whiskey Creek Shellfish Hatchery;
Thompson,Peter(peter.a.Thompson@csiro.au)-CSIRO Australia;
Wikfors,Gary A.(Gary.Wikfors@noaa.gov)-NOAA fisheries;
Langdon,Chris(Chris.Langdon@oregonstate.edu)-Oregon State University;
Samain,Jean Francois(jfsamain@ifremer.fr)-IFREMER, France;
Boudry,Pierre(Pierre.boudry@ifremer.fr,)-IFREMER, France;
Guo,Ximing(xguo@hsrl.rutgers.edu)-Rutgers University;
Davis,Chris(cdavis@midcoast.com)-Penaquid Oyster Co.-University of Maine;
Cheney,Dan(cheney@pacshell.org)-Pacific Shellfish Institute;
Huvet,Armand(ahuvet@ifremer.fr)-IFREMER, France;
Davis,Joth(jdavis@mindspring.com)(Tayloresearch@yahoo.com)-Taylor Shellfish;
Rawson,Paul(prawson@maine.edu)-University of Maine;
Elston,Ralph(Ralph@aquatechnics.com)-Aquatechnics

Brief Summary of Minutes

Accomplishments

The selective breeding program at The University of Maine produced seed oysters for three improved stocks of the eastern oyster (Rutgers NEH line, University of Maine Flowers Select, and Frank M. Flowers JOD resistant lines). The seed oysters were deployed at cold-water sites throughout coastal Maine in August of 2003 and the site-specific relative growth, survival and yield for each line is being monitored until the fall of 2005. This project in Maine mirrors a similar project that Dr. Ximing Guo presented at WCC-099 meetings for the past several years. Dr. Guo has been monitoring the performance of these lines in southern New England where water temperatures are warmer and the diseases MSX and Dermo are more prevalent than in Maine. Because the grow-out conditions in Maine are typically much colder than in southern New England we look forward to comparing the final datasets.<br /> <br><br /> <br>The Guo lab at the Haskin Shellfish Research Laboratory, Rutgers University, is also focusing on the genetic improvement of the Eastern Oyster. The Rutgers disease-resistant strain was crossed with the Frank M. Flowers fast-growing variety, and the resulting hybrids provided the highest yield exhibiting a combination of disease-resistance and superior growth. <br /> <br><br /> <br>All-triploid eastern oysters were produced from Rutgers disease-resistant strains using tetraploids. Triploidy is expected to enhance the growth rate of the Rutgers disease-resistant stock. The triploids are now being evaluated in the field. Work is ongoing on the mapping of disease-resistance genes in the eastern oyster.<br /> <br><br /> <br>In the Langdon laboratory, a collaborative experiment with French colleagues was carried out in which the effect of diet composition on the immune response of Pacific Oysters was determined. The project involved university researchers in Oregon, Virginia and IFREMER, Brest, France. All researchers were participants in WCC-99. <br /> <br><br /> <br>The Molluscan Broodstock Program benefited from comments made at last year?s WCC-99 meeting and resulted in development of a rotational breeding program to reduce inbreeding depression. This breeding program has been established and will be evaluated again next year.<br /> <br><br /> <br>Molecular work is underway at VIMS and Rutgers to develop various RFLP, AFLP and microsatellites to distinguish oyster species from one another. Three putative QTLs have been identified which are associated with disease resistance.<br /> <br><br /> <br>For more detail on these and other accomplishments, please refer to the Minutes of the February 29, 2004, meeting of WCC-099, found associated with this document on NIMMS.

Publications

Li,L. and X. Guo. 2004. AFLP-based genetic linkage maps of the Pacific oyster Crassostrea gigas Thunberg. Marine Biotechnology 6:26-36;<br /> <br><br /> <br> <br /> <br>Jackson, D.L., B.W. MacDonald, B. Vercaemer, X. Guo, A. Mallet and E.L. Kenchinton. 2003. Investigations with Triploid Atlantic Sea Scallops, Placopecten magellanicus, at the Bedford Institute of Oceanography, 2000-2003. Can. Tech. Rep. Fish. Aquat. Sci. 2460: v+ 48p.;<br /> <br><br /> <br> <br /> <br>Guo, X. S.E. Ford, G. DeBrosse, R. Smolowitz and I. Sunila. 2003. Breeding and evaluation of eastern oyster strains selected for MSX, Dermo and JOD resistance. J. Shellfish Res., 22(1):333-334.<br /> <br> <br /> <br>Wang, Y. and X. Guo. 2003. Chromosomal mapping of ribosomal RNA genes and telomeric repeats in zhikong and bay scallops. J. Shellfish Res., 22(1):360.<br /> <br><br /> <br>Langdon, C.J., Evans, F., Jacobson, D. and Blouin, M. 2003, Yields of cultured Pacific oysters Crassostrea gigas Thunberg improved after one generation of selection. Aquaculture, 220:227-244.<br /> <br><br /> <br>Brake, J., F. Evans and C. Langdon. 2003. Evidence for genetic control of pigmentation of shell and mantle edge in selected families of Pacific oysters, Crassostrea gigas. Aquaculture, 229:89-98.<br /> <br><br /> <br>Brake, J. F. Evans and C. Langdon. 2003. Is beauty in the eye of the beholder? Development of a simple method to describe desirable shell shape for the Pacific oyster industry. J. Shellfish Research, 22:767-771.<br /> <br><br /> <br>Evans, F., S. Matson, J. Brake and C. Langdon. 2003. The effects of inbreeding on performance traits of adult Pacific oysters (Crassostrea gigas). Aquaculture, 230:89-98.<br /> <br><br /> <br>Rawson, P.D., A. McGowen, and C. Bartlett. 2004. Temporal Variation in the Settlement of Blue Mussels (Mytilus edulis C. Linnaeus, 1758 and M. trossulus Gould 1850) in Eastern Maine. Journal of Shellfish Research, (in press).

Impact Statements

1. WCC-99 continues to be a very important forum for interaction and exchange of ideas among a wide range of shellfish geneticists from the US and abroad. Researchers from breeding programs in France, Australia and New Zealand are active participants. We also have strong input from researchers on the East Coast involved in restoration of the Eastern oyster in the Chesapeake Bay.
2. Participation in WCC-099 has had a direct impact on the research conducted on oyster culture in Maine. It has allowed the building of collaborations with selective breeding programs in other states and to map out approaches for improving the genetic resources available to the oyster research community, approaches that will significantly benefit not only research in Maine but the research in other states, as well.

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Report Information

Participants

Allen,Standish(ska@vims.edu)-Virginia Inst. Marine Science;
Boudry,Pierre(pboudry@ifremer.fr)IFREMER;
Camara,Mark-Oregon State University-USDA-Agric. Res. Serv.;
Carlsson,Jens(jc@vims.edu)-Virginia Inst. Marine Science;
Chiarri-Gomez,Marta(gomezchi@uri.edu)-Univ. of RI;
Cordes,Jan(jfcordes@vims.edu)-Virginia Inst. Marine Science;
Cunningham,Charles(cunnin@musc.edu)-Medical Univ. of South Carolina;
Davis,Chris(cdavis@midcoast.com)-Univ. of Maine;
DeBrosse,Greg(debrosse@rci.rutgers.edu)-Rutgers Univ.;
DeGremont,Lionel(ldegremo@vims.edu)-Virginia Inst. Marine Science;
DeRosa,Andy(derosa@agcenter.lsu.edu)-Louisiana State University;
Eudeline,Benoit(benoite@talorshellfish.com)-Taylor United;
Evans,Ford(ford.evans@oregonstate.edu)-Oregon State;
Gaffney,Patrick(pgaffney@udel.edu)-Univ. Delaware;
Guo,Ximing(xguo@hsrl.rutgers.edu)-Rutgers Univ.;
Hedgecock,Dennis(dhedge@usc.edu)-Univ. Southern California;
Itoh,Naoki(nitoh@lsu.edu)-Louisiana State University;
Jenny,Matt(jennymj@musc.edu)-Medical Univ. of South Carolina;
Karney,Rick(musg@capecod.net)-Martha's Vineyard Shellfish;
Langdon,Chris(chris.langdon@oregonstate.edu)-Oregon State;
Leavitt,Dale(dleavitt@rwu.edu)-Roger Williams Univ.;
Lindell,Scott(slindell@mbl.edu)-Marine Biological Laboratory;
Merritt,Don(meritt@hpl.umces.edu)-Univ. Maryland/ Sea Grant;
Rawson,Paul(prawson@maine.edu)-Univ. of Maine;
Reece,Kimberly(kreece@vims.edu)-Virginia Inst. Marine Science;
Ristow,Sandra(ristow@wsu.edu)-Washington State Univ.;
Roberts,Steve(sroberts@mbl.edu)-Marine Biological Laboratory;
Surier,Amadine(musg@capecod.net)-Martha's Vineyard Shellfish;
Taris,Nicolas(ntaris@ifremer.fr)-IFREMER;
Villamil,Luisa(villamil@mail.uri.edu)-University of RI;
Wilbur,Ami(wilbura@uncw.edu)-Univ. of NC-Wilmington;

Brief Summary of Minutes

Please see the attached.

Accomplishments

Accomplishments of WERA-099:<br /> <br /> (1) The twenty year Western Regional Aquaculture Consortium oyster breeding project on C. gigas has been completed and the resultant improved lines given to Taylor United for further improvements. That work has yielded some better performing more commercially desirable lines.<br /> <br /> (2) An oyster hepatopancreas microarray containing genes for both C. gigas and C. virginica as well as some genes from one of the oyster pathogens, Haplosporidium nelsoni, has been developed by members of WERA -099 at the Medical University of South Carolina. It is widely available for the oyster breeding community to assist in studies of growth, physiology, disease states including juvenile oyster disease, and in charting the immune system of oysters.<br /> <br /> (3) With the advent of DNA typing and microarrays, WERA-099 participants are now cognizant that morphological characteristics do not necessarily determine the species of oyster; and that in order to perform intelligent breeding schemes and to perform restoration with particular stocks, scientists will have to establish a reference library of DNA from what are considered the early forms of each oyster species.<br /> <br /> (4) Members of WERA-099 have made a decision to take advantage of the National Animal Germplasm Program, Fort Collins CO and place specimens of valuable lines in the bank. Members of the group also agreed to donate reference DNA to that repository from their important lines.<br /> <br /> (5) Members of WERA-099 are preparing a joint multi-state proposal for the sequencing of the genome of the oyster. The work began at the WERA-099 meeting in Philadephia, April 10.<br /> <br /> <br /> <br />

Publications

Peer Reviewed Publications:<br /> <br /> Committee on Nonnative Oysters in the Chesapeake Bay J. Anderson (Co-Chair), D. Hedgecock (Co-Chair), M. Berrigan, K. Criddle, W. Dewey, S. Ford, P. Goulletquer, R. Hildreth, M. Paolisso, N. Targett, R. Whitlatch. 2004. Nonnative Oysters in the Chesapeake Bay. National Research Council of the National Academies. The National Academies Press, Washington, D.C. 325 pp.<br /> <br /> Hubert, S., and D. Hedgecock. 2004. Linkage maps of microsatellite DNA markers for the Pacific oyster Crassostrea gigas. Genetics 168:351-362.<br /> <br /> Hedgecock, D., G. Li, S. Hubert, K. Bucklin, and V. Ribes. 2004. Widespread null alleles and poor cross-species amplification of microsatellite DNA loci cloned from the Pacific oyster (Crassostrea gigas). Journal of Shellfish Research 23:379-385.<br /> <br /> He, Y., Dong, Q., Tiersch, T.R., and Devireddy, R.V. 2004. Variation in the membrane transport properties and predicted optimal rates of freezing for spermatozoa of diploid and tetraploid Pacific oyster Crassostrea gigas. Biology of Reproduction 70:1428-1437.<br /> <br /> Dong, Q., Eudeline, B., Huang, C., and Tiersch, T.R. 2005. Standardization of photometric measurement of sperm cell concentration from diploid and tetraploid Pacific oysters, Crassostrea gigas (Thunberg). Aquaculture Research 36:86-93.<br /> <br /> Dong, Q., Eudeline, B., Huang, C., Allen, S.K., and Tiersch, T.R. 2005. Commercial-scale sperm cryopreservation of diploid and tetraploid Pacific oysters, Crassostrea gigas. Cryobiology 50:1-16.<br /> <br /> Reece, K.S., Ribeiro, W.L., Gaffney, P.M., Carnegie, R.B., and Allen, S.K. Jr. (2004) Microsatellite marker development and analysis in the eastern oyster, Crassostrea virginica: Confirmation of null alleles and non-Mendelian segregation ratios. J. Heredity 95:355-361.<br /> <br /> Huvet, A., Herpin, A., Dégremont, L., Labreuche, Y., Samain, J.F., and Cunningham, C. (2004) The identification of genes from the oyster Crassostrea gigas that are differentially expressed in families exhibiting opposed susceptibility to summer mortality. Gene 343:211-220.<br /> <br /> Kim, H-W, Mykles, D.L., Goetz, F.W., Roberts, S.B. (2004) Characterization of an invertebrate myostatin homologue from the bay scallop, Argopecten irradians. BBA Gene Structure and Expression. 1679(2):174-9 <br /> <br /> Roberts, S.B. (2004) Lab Studies: Genes Involved with Growth, Development of Bay Scallops. Global Aquaculture Advocate. 7(3): 55-56<br /> <br /> <br /> <br /> Publications in Review<br /> <br /> Camara, M.D., Carnegie, R. Allen, S.K. Jr. (in review) Outcrossing among commercial strains of the Northern Quahog, Mercenaria mercenaria: Survival growth, and implications for selective breeding. Submitted to Aquaculture.<br /> <br /> <br /> Zhang, Q, Allen, S.K., Jr., and Reece, K.S. (in press) Genetic variation in wild and hatchery stocks of the Suminoe oyster (Crassostrea ariakensis) assessed by PCR-RFLP and microsatellite markers. Mar. Biotechnol.<br /> <br /> <br /> Herpin, A., Lelong, C., Becker, T., Favrel, P., Cunningham, C. Structural and functional evidence for a type 1 TGF-beta sensu stricto receptor in the lophotrochozoan Crassostrea gigas. Mechanisms of Development (in press).<br /> <br /> Herpin, A., Becker, T., Rosa, F.M., Favrel, P., Cunningham, C. Molecular and functional characterization of a new Tolloid like protein from a lophotrochozoan organism suggests orthology to its vertebrate counterparts. Submitted to Evolution & Development.<br /> <br /> Herpin, A., Lelong, C., Becker, T., Favrel, P., Cunningham, C. Structural and functional evidence for a type 1 BMP receptor in the lophotrochozoan Crassostrea gigas. Submitted to Gene.<br /> <br /> Roberts, S.B., Romano, C., Gerlach, G. (2005) Characterization of EST derived SSRs from the bay scallop, Argopectens irradians. Molecular Ecology Notes. In press<br /> <br /> Abstracts<br /> Hedgecock, D., G. Li, and M.-L. Voigt. 2004. Mapping heterosis QTL in the Pacific oyster Crassostrea gigas. http://www.intl-pag.org/12/abstracts/W06_PAG12_19.html<br /> <br /> Lundqvist, M.L., M.J. Jenny, G.W. Warr, P.S Gross, G.R. Vasta, J.A.F. Robledo, Z.J. Liu, J. Tomkins, G.C. Fang, C. Saski, M. Gomez-Chiarri, J.-M. Escoubas, E. Bachere, P. Roche, D. Hedgecock, and R.W. Chapman. 2004. Current state of oyster (Crassostrea) functional genomics resources. http://www.intl-pag.org/12/abstracts/P5o_PAG12_735.html<br /> <br /> Presentations<br /> <br /> Langdon, C. 2004. Challenges for Aquaculture in the 21st century. Seminar speaker, School of Fisheries, University of Washington.<br /> <br /> Langdon, C. 2004. Breeding better oysters. Seminar. Plymouth Marine Laboratories, U.K.<br /> <br /> Langdon, C., Evans, F., Brake, J., Matson, S. 2004. Improvement in yields of the Pacific oyster Crassostrea gigas by means of family selection. 96th annual meeting of the National Shellfisheries Association, March 1-5, 2004, Honolulu, USA.<br /> <br /> Evans, F., Langdon, C., 2004. Effects of genotype-environment interactions on yield of Pacific oyster (Crassostrea gigas) families in the Pacific Northwest. 58th Annual Meeting of the National Shellfisheries Association Pacific Coast Section and Pacific Coast Shellfish Growers Association, Tacoma, WA (USA), Oct. 12-14, 2004.<br /> <br /> Evans, F., Langdon, C., 2004. The effects of nursery environment on adult performance traits in Pacific oysters Crassostrea gigas. 96th annual meeting of the National Shellfisheries Association, March 1-5, 2004, Honolulu, USA<br /> <br /> Reece, K.S. A Non-native Oyster for Chesapeake Bay? Seminar presentation at Oregon State University February 24, 2004. <br /> <br /> Reece, K.S. and Burreson, E.M. The Role of Molecular Biology in International Regulation: Shellfish Health. Invited Workshop Presentation. Aquaculture Biotechnology Workshop Sponsored by St. Andrews Biological Station, Fisheries and Oceans Canada (DFO), Aquaculture Association of Canada Workshop. May 11-12, 2004.<br /> <br /> Reece, K.S. A Non-native Oyster for Chesapeake Bay; Penny-wise, pound-foolish? Seminar presentation at Juniata College. June 30, 2004. <br /> <br /> Reece, K.S. Crassostrea ariakensis for Chesapeake Bay? Species Identification and Crassostrea spp. Phylogeny, Diseases in the Native Range. Seminar Presentation at Rutgers University, November 22, 2004.<br /> <br /> Luckenbach, M., Carnegie, R., Coen, L.D., Paynter, K., Jr. and Reece, K.S. (2004) Observations on the ecology of Crassostrea ariakensis in its native range. International Conference on Shellfish Restoration. November 17-20, 2004, Charleston, SC.<br /> <br /> Cordes, J.F. and Reece, K.S. (2004) Genetic identification of oyster species based on restriction fragment length polymorphism (RFLP) analysis of two molecular markers amplified using the polymerase chain reaction (PCR). International Conference on Shellfish Restoration. November 17-20, 2004, Charleston, SC.<br /> <br /> Carnegie, R.B., Scott, G.P., Johnson, K., Ragone Calvo, L.M., Camara, M.D. and Reece, K.S. (2004) Development of a microsatellite marker set for the hard clam Mercenaria mercenaria. Aquaculture 2004, March 1-5, Honolulu HI.<br /> <br /> Evans, F., Langdon, C., 2004. Effects of genotype-environment interactions on yield of Pacific oyster (Crassostrea gigas) families in the Pacific Northwest. 58th Annual Meeting of the National Shellfisheries Association Pacific Coast Section and Pacific Coast Shellfish Growers Association, Tacoma, WA (USA), Oct. 12-14, 2004.<br /> <br /> Evans, F., Langdon, C., 2004. The effects of nursery environment on adult performance traits in Pacific oysters Crassostrea gigas. 96th annual meeting of the National Shellfisheries Association, March 1-5, 2004, Honolulu, USA<br /> <br /> Langdon, C, Evans, F., Brake, J., Matson, S. 2004. Improvement in yields of the Pacific oyster Crassostrea gigas by means of family selection. 96th annual meeting of the National Shellfisheries Association, March 1-5, 2004, Honolulu, USA.<br />

Impact Statements

1. The oyster breeding work on C. gigas begun by the Western Regional Aquaculture Consortium about 20 years ago has been completed and the resultant improved lines turned over to Taylor United, Shelton WA. That work has yielded several better performing, more commercially desirable lines.
2. An oyster hepatopancreas microarray containing genes for both C. gigas and C. virginica as well as some genes from one of the pathogens of oyster, Haplosporidium nelsoni, has been developed by members of WERA -099 residing at the Medical University of South Carolina. The microarray is widely available for the oyster breeding community to assist in studies of growth, physiology, disease states including juvenile oyster disease, and in charting the immune system of oysters.
3. With the advent of DNA typing and microarrays, WERA-099 participants are now cognizant that morphological characteristics do not necessarily determine species and that in order to perform intelligent breeding schemes and to perform restoration with particular stocks, oyster scientists will have to establish a reference library of DNA from what are considered the early forms of each species.
4. The group decided to take advantage of the preservation capabilities of the National Animal Germplasm Program, Fort Collins CO and collaborate to place specimens of valuable lines of oysters in the bank. The members of the group also agreed to donate reference DNA to that repository from their important lines.
5. The group is preparing a joint multi-state proposal for the sequencing of the genome of the oyster. The work began at the WERA-099 meeting in Philadephia, April 10.

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Report Information

Participants

Joth Davis - Taylor Resources, Inc. Washington (jdavis@bainbridge.net);
Jason Curole - University of Southern California (jcarole@usc.edu);
Mark Camara - USDA-ARS, Hatfield Marine Sciences Center (mark.camara@oregonstate.edu);
Paul Lange - Oregon State University (paul.lange@oregonstate.edu);
Alan Barton - Oregon State University, Molluscan Broodstock Program (alan.barton@oregonstate.edu);
David A. Stick - Oregon State University (david.stick@oregonstate.edu);
Gregory DeBrosse - Rutgers Haskin Shellfish Laboratoy (debrosse@rci.rutgers.edu);
Ximing Guo - Rutgers University (xguo@hsrl.rutgers.edu);
Chris Davis - University of Maine (cdavis@midcoast.com);
Brent Vadapalas - University of Washington (brentv@u.washington.edu);
Kristi Straus - University of Washington (kmstraus@u.washington.edu);
Marta Gomez-Chiarri - University of Rhode Island (gomezchi@umd.edu);
Matthew Hare - University of Maryland (matthare@umd.edu);
Rick Karney - Martha's Vineyard Island Shellfish Group (mvsg@adelphia.net);
Dale Leavitt - Roger Williams University (dleavitt@rwu.edu);
Amandine Surier - Martha's Vineyard Island Shellfish Group (amandine_surier@hotmail.com);
Christopher Langdon - Oregon State University (chris.langdon@oregonstate.edu);
Pierre Boudry - IFREMER (La Tremblade) France (pboudry@ifremer.fr);
Jean Francois Samain - IFREMER (Brest) France (jfsamain@ifremer.fr);
Lionel Degremont - ABC - Virginia Institute of Marine Sciences (ldegremo@vims.edu);
Kim Reece - Virginia Institute of Marine Sciences (kreece@vims.edu);
Jan Cordes - Virginia Institute of Marine Sciences (jfcordes@vims.edu);
Ryan Carnegie - Virginia Institute of Marine Sciences (carnegie@vims.edu);
Jens Carlsson - Virginia Institute of Marine Sciences (jc@vims.edu);
Dennis Hedgecock - University of Southern California (dhedge@usc.edu);
Sandra Ristow - Washington State University - Pullman WA (ristow@wsu.edu);
Don Meritt - University of Maryland -UMCES (merit@hpl.umces.edu);
Ford Evans - Oregon State University (ford.evans@oregonstate.edu);
Ami Wilbur - University of North Carolina - Willmington (Wilbur@uncw.edu);
Stan Allen - Virginia Institute of Marine Sciences (ska@vims.edu);
Ray Ralonde - Alaska Sea Grant - University of Alaska (afrlr@uaa.alaska.edu);
Steven Roberts - Marine Biological Laboratory - Woods Hole (sroberts@mbl.edu);
Scott Lindell - Marine Biological Laboratory - Woods Hole (slindell@mbl.edu);
Arpita Chaudhary - NOAA (arpita.chaudhary@noaa.gov);
Dan Cheney - Pacific Shellfish Institute (psi@pacshell.org);
Eric Powell - Haskin Shellfish Laboratory, Rutgers (eric@hsrl.rutgers.edu);
Lingling Wang - Haskin Shellfish Laboratory - Rutgers (lingling@hsrl.rutgers.edu);
JeongHo Lee - Haskin Shellfish Laboratory - Rutgers (jhlee@hsrl.rutgers.edu);

Brief Summary of Minutes

Accomplishments

<br /> <br /> 1. A key finding from several laboratories determining the genetic basis for heterosis and crossbreeding in Pacific Oysters is that superior hybrid performance relative to inbred performance relies on a combination of additive and dominance effects.<br /> <br /> <br /> 2. A PCR/RFLP identification key based on the ITS-1 and COI gene regions for Indo Pacific Crassostrea species is able to differentiate between ariakensis and other species.<br /> <br /> <br /> 3. Five new microsatellite markers have become useful in the preservation of genetic diversity in wild hard shell clams (Quahogs) and distinguishing them from cultured clams.<br /> <br /> <br /> 4. In Maine, a collaboration has emerged between the University and ten farms which helps to support the oyster breeding program for that region.<br /> <br /> <br /> 5. An effort is proceeding in Virginia to foster selective breeding in order to layer MSX resistance atop dermo resistance.<br /> <br /> <br /> 6. Several important outcomes of immune response in oysters have been delineated. Nitric oxide production is intimately involved in parasite control in oysters. Histone H4 is also involved in response to infection.<br /> <br /> <br />

Publications

Matson, S., Langdon, C., Evans, S., 2006. Specific pathogen free culture of the Pacific oyster (Crassostrea gigas) in a breeding research program: growth effects and remediation. Aquaculture 253, 475-484.<br /> <br /> Camara, M.D., Griffith, S.M., Evans, S. 2005. Can selective breeding reduce the heavy metals content of Pacific oysters (Crassostrea gigas), and are there trade-offs with growth or survival? J. Shellfish. Res. 24, 979-986.<br /> <br /> Hedgecock, D., P. M. Gaffney, P. Goulletquer, X. Guo, K. Reece, and G. W. Warr. 2005. The case for sequencing the Pacific oyster genome. Journal of Shellfish Research 24:429-441.<p><br /> <br /> Curole, J. P., and D. Hedgecock. 2005. Estimation of preferential pairing rates in second-generation autotetraploid Pacific oysters (Crassostrea gigas). Genetics 171:855-859.<p><br /> <br /> Yamtich, J., M.-L. Voigt, G. Li, and D. Hedgecock. 2005. Eight microsatellite loci for the Pacific oyster Crassostrea gigas. Animal Genetics 36:524-526.<p><br /> <br /> Pace, D. A., A. G. Marsh, P. Leong, A. Green, D. Hedgecock, and D. T. Manahan. 2006. Physiological bases of genetically determined variation in growth of bivalve larvae (Crassostrea gigas). Journal of Experimental Marine Biology and Ecology, in press.<p><br /> <br /> Pubished abstracts:<br /> Curole, J. P., and D. Hedgecock. 2004. High frequency of SNPs in the Pacific oyster genome. http://www.intl-pag.org/13/abstracts/PAG13_W026.html<p><br /> <br /> Curole, J. P., and D. Hedgecock. 2005. Genetic mapping of candidate loci for growth heterosis in the Pacific oyster. http://www.intl-pag.org/14/abstracts/PAG14_W186.html<p> <br /> <br />

Impact Statements

1. WERA-099 blends an excellent group of researchers, industry representatives and extension personnel in a discussion of the current problems facing the oyster industry. At the meeting there are intense discussions of breeding systems, culture techniques, nutrition and disease problems. Many collaborative projects have emerged as a result of these discussions.
2. WERA-099 enhances international collaboration. Researchers from France, New Zealand and Australia regularly attend the meetings.
3. WERA-099 promotes graduate education. Graduate students regularly attend these meetings, present their findings and interact with faculty from other universities. They access information that is applicable to their present and future studies. Occasionally, students are matched with post-doctoral fellowships as a result of interactions at the meeting.

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