Ostreid herpesvirus 1 (OsHV-1) is the only member of the Herpesviridae that has an invertebrate host and is associated with sporadic mortality in the Pacific oyster (Crassostrea gigas) and other bivalve species. Cryo-electron microscopy of purified capsids revealed the distinctive T=16 icosahedral structure characteristic of herpesviruses, although the preparations examined lacked pentons. The gross genome organization of OsHV-1 was similar to that of certain mammalian herpesviruses (including herpes simplex virus and human cytomegalovirus), consisting of two invertible unique regions (U L , 167?8 kbp; U S , 3?4 kbp) each flanked by inverted repeats (TR L /IR L , 7?6 kbp; TR S /IR S , 9?8 kbp), with an additional unique sequence (X, 1?5 kbp) between IR L and IR S . Of the 124 unique genes predicted from the 207 439 bp genome sequence, 38 were members of 12 families of related genes and encoded products related to helicases, inhibitors of apoptosis, deoxyuridine triphosphatase and RING-finger proteins, in addition to membrane-associated proteins. Eight genes in three of the families appeared to be fragmented. Other genes that did not belong to the families were predicted to encode DNA polymerase, the two subunits of ribonucleotide reductase, a helicase, a primase, the ATPase subunit of terminase, a RecB-like protein, additional RING-like proteins, an ion channel and several other membrane-associated proteins. Sequence comparisons showed that OsHV-1 is at best tenuously related to the two classes of vertebrate herpesviruses (those associated with mammals, birds and reptiles, and those associated with bony fish and amphibians). OsHV-1 thus represents a third major class of the herpesviruses. INTRODUCTIONViruses are assigned to the family Herpesviridae on the basis of morphological criteria and have been identified in a wide range of vertebrates and one invertebrate, the Pacific oyster, Crassostrea gigas (Minson et al., 2000). The vertebrate herpesviruses fall into two major phylogenetic groups. Those in the first group have mammalian or avian hosts and are classified into three subfamilies (Alphaherpesvirinae, Betaherpesvirinae and Gammaherpesvirinae) that share extensive genetic relationships . Reptilian herpesviruses probably also belong among the Alphaherpesvirinae (Nigro et al., 2004;Quackenbush et al., 1998;Une et al., 2000;Yu et al., 2001). Viruses in the second group infect amphibians or bony fish and again are interrelated (Bernard & Mercier, 1993;Davison, 1998; Davison et al., 1999). Genetic evidence for a common evolutionary origin for the two groups is tenuous, however, since not a single herpesvirus-specific gene is detectably conserved in both. The only completely sequenced lower vertebrate herpesvirus, channel catfish virus (CCV), does share a few genes with the higher vertebrate group, but these have counterparts in other organisms (Davison, 1992). The conserved gene that comes closest to being herpesvirus specific encodes the putative ATPase subunit of the terminase, an enzyme complex involved in p...

Concomitant sporadic high mortalities were reported in France in May 1994 among batches of hatchery-reared larval Pacific oysters Crassostrea gigas and European flat oysters Ostrea edulis in 2 hatcheries, and in June and July 1994 among batches of cultured spat of both species in a shellfish nursery. Histological observation showed the presence of cellular abnormalities in moribund animals. Transmission electron microscopy revealed the presence of herpes-like virus particles in infected larvae and spat of both oyster species. This is the first description of a herpes-like virus infection in larval O. edulis. Viruses observed in diseased larvae and spat of both species are similar with respect to ultrastructure and morphogenesis. They were detected simultaneously in C. gigas and O. edulis larvae and spat, indicating possible interspecific transmission. Moreover, these viruses are associated with high mortality rates in both oyster species. An electron microscopic examination revealed hemocytes with condensed chromatin and extensive perinuclear fragmentation of chromatin. These data suggest that herpes-like viruses infecting oysters may induce apoptosis in oyster hemocytes. KEY WORDS: Herpes-like virus · European flat oyster · Ostrea edulis · Pacific oyster · Crassostrea gigas · Oyster mortality · Concomitant infection · Virus replication · Apoptosis Resale or republication not permitted without written consent of the publisherDis Aquat Org 42: [173][174][175][176][177][178][179][180][181][182][183] 2000 viral particles reported by Hine et al. (1992) in New Zealand and the virus-like particles described by Comps & Cochennec (1993) among French cultured European flat oyster spat.We describe for the first time a herpes-like virus infecting European flat oyster larvae and concomitant herpes-like virus infections associated with high mortalities among hatchery-reared larvae and nurserycultured spat of both Crassostrea gigas and Ostrea edulis during the summer of 1994 in France. We performed an ultrastructural comparative study between both virus-like particles found respectively in Pacific oysters and European flat oysters and compared them to viruses belonging to the Herpesviridae family. MATERIALS AND METHODS Specimens.Larval Ostrea edulis and Crassostrea gigas, 6 to 10 d old, 80 to 150 µm in size, were collected in May 1994 from 2 hatcheries located in Bourgneuf Bay (Vendée, France). Nursery-cultured spat of C. gigas and O. edulis, 3 to 5 mo old, were also collected from Bourgneuf Bay in June and July 1994.Light microscopy. A total of 197 European flat oyster spat and 196 Pacific oyster spat were collected for light microscopical examination. Moribund cultured spat of both species, 3 to 5 mo old, were removed from the shell and, sagittally sectioned; then half of each specimen was fixed in Davidson's fixative for light microscopic examination and the other half in Carson's fixative for transmission electron microscopic analysis. After 48 h fixation in Davidson's fluid, samples were dehydrated using an ...

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Summary. Pacific oyster, Crassostrea gigas, is the most economically important specie to the world shellfish breeding. It is important to note that infectious diseases, particularly viruses, may be hazardous for the C. gigas live-stocks. The study of these viral diseases and the development of diagnosis method need the establishment of in vitro methods for viral multiplication. As no oyster cell line is available actually, we have developed a procedure for primary culture of heart cells which could enable to study molluscan viruses in vitro, and could also provide a diagnosis method based on the search of eventual cytopathogen viral effects. Cells from C. gigas ventricle of heart were dissociated by trypsin-EDTA treatment and the mechanical action of a Dounce type homogeneizer. The cells were inoculated in previously poly-D-lysin coated flasks. The optimised culture medium was L-15 (Leibovitz) prepared three fold concentrated, then diluted half with sea water, this mixture was supplemented with 10% FCS and 5% C. gigas hemolymph. Different cell types could be identified by transmission electron microscopy analysis, as mostly cardiomyocytes, fibroblast-like cells and pigmented cells, but also haemocytes were present in the cultures.