Contribution à l'étude de virus de mollusques marins apparentés ...

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HERPESVIRUSES ASSOCIATED WITH MORTALITIES AMONG PACIFIC OYSTER, IN FRANCE 739 viruses. However sorne analysed animais exhibited ab nor- 1 mal nulei and condensed chromatin without virus detection (Table 1). No virus was observed among healthy wild-type larvae and healthy or affected by moderate mortality cultured young oysters (Table 1). Discussion The virions described in this report ressemble hespesviruses in morphological characteristics, in cellular locations and in size range [1 8, 15, 17]. In addition, the fibrils spanning the space from the core to the inner surface of the capsid are similar to the arrangement in herpesviruses [1 J, 17] . The herpes-li ke virus associated with mortalities among hatchery-reared larval Pacific oysters, C. gigas, seems to Origine of simple Dale of Nature of sample I nlmals Loire-Atlantique (hatchery-reared) 24.06.92 L .. 13.07.92 L .. 15.07.92 L .. 29.07.92 L Charente-Maritime (wild-type) 08.07.92 L .. 29.07.92 L .. 29.07.92 L Charente-Maritime (halchery-reared) .. .. .. 25.06.93 12.07.93 14.07.93 19.07.93 L L L L loire-AtianUque .. Charente-Maritime 06.07.93 25. 10.93 YS YS (Oléron) .. Charente-Maritime 08.07.93 17.09.93 YS YS (Oléron) Charente-Maritime 08.07.93 YS (Boyal
740 RENAULT (T.) ET COLLABORATEURS Size (fi . 50) Capalda Nucloocapsld6 Envoloplng , Enveloped virions virions Bh J.Onm 75" 3.0 nm 991: 0.0 nm 122 ± B.O nm Nucleus + + Perfnuclelr space + Cvtoplaom + + + Denie bodies + + Cytoplumle + + vellel". Extrllcellul.r + + TABLE Il. - Occurrence and sile of diffcrent stages of replicalion of oysler herpes-like virus in dirfere"1 cell parts of Crassostrea gigas larvae. Cap. Ida Nucloocapslda Enveloplnu Enveloped virions Ylrion. 51%0( ... 50) 81 j: 3.0 nm 75.13.0 nm 100±9.0nm 121 ... 7.0 nm Nucl.u. + + Perlnuclear .pace + Cytopillm + + + Denllbodln + + Cytopl •• mlc + + vellel .. Exr'Clllular + ! TABLE III. - Occurrence and .size of dirfcrent stages of replicalion of oyster herpes-like virus in diffcre"t cell parts of young oysters, Crassastra gigas. such similarities are not sufficient on a taxonomic point. Details of replication are variable within subfamilies [8] and vary with cell type infected [8, 21 ], time after infection [18] and virus strains [3] . Nevertheless, the nueleic acid nature of these viruses should be confirmed by chemical analysis. Several reports of herpes-like viruses from marine vertebrates or invertebrates are found in the literature [2, 12, 13, 19], including herpes-type viruses in oysters [l, 7, 10, 14, 16]. The morphological characteristics of the nucleocapsid of the viruses described in this sludy are c10sed to those of the viruses reported among Pacific oyster, C. gigas, larvae in french hatcheries [16] and in New Zealand [1 4]. The nueleocapsids are similar in dimension to those detected in European flat oyster, Ostrea edulis, by COMPS and COCHENNEC [7], in Pacific oyster, C. gigas, by NICOLAS et al. [15] and in Arnerican oyster, C. virginiea, by FAR LEY et al. [10], but at 70-75 nm, they are smaller than the 97 nm of nueleocapsids observed by HINE et al. [14], among C. gigas in New Zealand (Table IV). Moreover, mature virions described in trus study are smaller than the 160 to 180 nm of enveloped paraspherical 83 partieles observed in O. edulis [7] and larger than the 90 nm of enveloped partieles reported among French Pacific oyster larvae [16] (fable IV). Mature partieles noted into Pacific oyster larvae in New Zealand [14] and in our report are similar in size (Table IV). However, partieles sizes are dependent on the technique of specimen preparation and measurement. Elevated temperature and crowding seem to increase the susceptibility of animais to herpes-like virus infections among Pacific oyster, C. gigas. Indeed mortalities associated with herpes-like viruses detection were only observed during summer in this report. Moreover, the deteclion of herpesviruses in arumals held at elevated temperatures among C. virginiea [10], the occurrence of herpesviruses in summer among C. gigasin Australia [1] and the observation of mortalities in mid-summer among the same species reported in New Zealand [14], in France [16] and in this study, suggest ostreids herpesviruses may only develop and induce mortalities at elevated temperatures, particularly under stressed conditions. So high temperatures appear to favor the spread of the infections or activation of viruses from an occult 10 an overt phase or both.
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1 1 1 1 Année 1995 UNIVERSITE DE B
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à mes parents à Stéphane
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1 1 ) TABLE DES MATIERES INTRODUCTI
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3 - Essais d'infection expérimenta
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INTRODUCTION
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d'approche pour contrôler les mala
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1 1 1 1 1 1 1 1 j Un premier chapit
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DONNEES BIBLIOGRAPHIQUES 1 - Rappel
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1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
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et Wray, 1952 ; Mackin, 1962 ; Burr
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1 1 1 1 1 1 1 1 des zones indemnes
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3 - Procaryotes Les mollusques biva
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Tableau 4 : Infections à Chlamydie
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cheptels de C. angulala, a conduit
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t t 1 1 1 1 t 1 1 t 1 1 t 1 1 t 1 1
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i 1 1 1 1 1 1 1 1 1 1 1 1 1 J 1 int
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1 1 1 1 1 1 1 ! 1 1 1 1 ! 1 1 1 1 1
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III - Caractéristiques principales
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1 1 1 1 1 1 1 1 1 1 1 J 3.2 - Entr
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Page 45 and 46: 1 1 1 1 1 1 1 1 1 1 1 PREAMBULE Pub
Page 47 and 48: Chapitre 1 : Caractérisation d'un
Page 49 and 50: l 1 1 1 1 1 1 1 1 1 1 1 1 PUBLICATI
Page 52 and 53: References AllIle . w .. SchlOl fc
Page 55 and 56: Platichtys flesus, plie, Pleuronect
Page 57: Figure 1 : Marquage de coupes de do
Page 60 and 61: CHAPITRE 2 Comparaison antigénique
Page 62 and 63: Ces électrophorèses ont égalemen
Page 64 and 65: 1\6 , HMW LMW Figure 1 : Caractéri
Page 66 and 67: a 2 3 4 5 6 7 8 9 10 --- - - - - Il
Page 68 and 69: Tableau 2 : Comparaison anti géniq
Page 70 and 71: Figure 4 : Coupe histologique d'hû
Page 72 and 73: Chapitre 1 : Mise en évidence et d
Page 74 and 75: Les virions observés chez les larv
Page 76: • •• Figure 2 : Cliché en mi
Page 81 and 82: 2 - Participation à la mise en év
Page 83 and 84: Figure 10 : Lésions histologiques
Page 85: Bu ll. Eur. Ass. Fi sh Palhal.. 14(
Page 89: 736 Material and methods Samples of
Page 95 and 96: 742 15. - MURP HY (F.A.) and K INGS
Page 97: Chapitre Il : Essais de reproductio
Page 103 and 104: 2 - Essais d'infection expérimenta
Page 107 and 108: CHAPITRE 3 Effet de la température
Page 109 and 110: 26°C), de détecter la présence d
Page 111 and 112: présentent des lésions histologiq
Page 113: Introduction The Pacific oyster, 'C
Page 117 and 118: (Table 1). 100% mortality in this g
Page 119 and 120: than at high temperature (25-26°C)
Page 121 and 122: Our plans for testing this la st hy
Page 123: LEGEND TO FIGURES Figure 1 : Origin
Page 129 and 130: CHAPITRE IV : Essais de propagation
Page 132 and 133: Figure 1 : Principales caractérist
Page 135 and 136: 2 - Essais d'infection de primocult
Page 137 and 138: (SVF) et/ou d'hémolymphe de Crassa
Page 139: 68 0.2 g/I, KCI, No. P 1300' 2.77 g
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72 2. Cousserans F, Bonami lR, Comp
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2.2. Essais d'infection de primocul
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Chapitre V : Mise au point d'un pro
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Du fait de la présence d'une coqui
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• " (1 - " 100nm 100nm Figures 1
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Chapitre VI : Mise au point de mét
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chromatine est rejetée en périph
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528 RM Le DeuN et al ,GD Fig 2. Imm
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1.2 - Recherche de séquences conse
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DJBE16 DJBE2L DJBECI DJBE16 DJBE2L
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1.2.2 - Alignement des séquences D
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viral, des spots d'intensité plus
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visualisées sur les dépôts d'ADN
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Figure 6 : Diagnostic de l'infectio
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CONCLUSION L'ensemble de ce travail
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séquences disponibles dans les ban
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REFERENCES BIBLIOGRAPHIQUES Ahne W.
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Cahour A., 1979. Mar/eilia refringe
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Darlington R.W. et Moss L.H., 1969.
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Girard M. et Hirth L., 1989b. Méth
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Johnson D.C. et Spear P.G., 1982. M
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Lemaster S. et Roizman B., 1980. He
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Ohba M., Kanda K. et Aizawa K., 199
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Read G.S. et Frenkel N., 1978. Herp
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Shuster C.L. et Hillman T.E., 1963.
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1994 : P. Maffart, R.M. Le Deuff, T
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days, according to Reed and Muench'
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detect a single infected cell in a
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Vet Res (1995) 26. 539·543 © Else
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Annexe 2 : Méthodes et techniques
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minutes, puis les coupes sont réhy
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2.1 - Préparation des solutions -
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2.6 - Coloration négative Lors des
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Gel de concentration 4% H 20 Tampon
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le tampon de transfert et superpos
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NaCI NaN) Ajuster le pH à 7 Tampon
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Epuisement des anticorps 1 - Utilis
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6 - Prélever la phase supérieure
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Tris-HCl pH8 1 ml Préparer extempo
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2 - Prélever 1 ml de tampon d'hybr
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