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D 1037 7 Nov 2002

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DISEASES OF AQUATIC ORGANISMS
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Dis Aquat Org
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REVIEW
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Spring viremia of carp (SVC) 74
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15 W. Ahne1, H. V. Bjorklund2, S. Essbauer 3,*, N. Fijan4, G. Kurath5, J. R. Winton5 77
16 78
1
17 Institute of Zoology, Fishery Biology and Fish Diseases, Faculty of Veterinary Medicine of the Ludwig Maximilians 79

PP: cf
18 University of Munich, Kaulbachstrasse 37, 80539 Muenchen, Germany 80
2
19 Orion Pharma Animal Health, PO Box 425, 20101 Turku, Finland 81
3
20 WHO-centre for Comparative Virology, Institute of Medical Microbiology, Infectious and Epidemic Diseases, 82
Faculty of Veterinary Medicine of the Ludwig Maximilians University of Munich, Veterinaerstr. 13, 80539 Muenchen, Germany
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Department of Biology and Pathology of Fish and Bees, Veterinary Faculty, University of Zagreb, Heinzelova 55,
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PO Box 190, 10000 Zagreb, Croatia
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Western Fisheries Research Center, 6505 NE 65th Street, Seattle, Washington 98115, USA
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ABSTRACT: Spring viremia of carp (SVC) is an important disease affecting cyprinids, mainly common
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carp Cyprinus carpio. The disease is widespread in European carp culture, where it causes significant
29 91
morbidity and mortality. Designated a notifiable disease by the Office International des Epizooties, SVC
30 92
is caused by a rhabdovirus, spring viremia of carp virus (SVCV). Affected fish show destruction of tissues
31 in the kidney, spleen and liver, leading to hemorrhage, loss of water-salt balance and impairment of im- 93
32 mune response. High mortality occurs at water temperatures of 10 to 17°C, typically in spring. At higher 94
33 temperatures, infected carp develop humoral antibodies that can neutralize the spread of virus and such 95
34 carp are protected against re-infection by solid immunity. The virus is shed mostly with the feces and 96
35 urine of clinically infected fish and by carriers. Waterborne transmission is believed to be the primary 97
36 route of infection, but bloodsucking parasites like leeches and the carp louse may serve as mechanical 98
37 vectors of SVCV. The genome of SVCV is composed of a single molecule of linear, negative-sense, 99
38 single-stranded RNA containing 5 genes in the order 3’-NPMGL-5’ coding for the viral nucleopro- 100
39 tein, phosphoprotein, matrix protein, glycoprotein, and polymerase, respectively. Polyacrylamide gel 101
electrophoresis of the viral proteins, and sequence homologies between the genes and gene junctions
40 102
of SVCV and vesicular stomatitis viruses, have led to the placement of the virus as a tentative member
41 103
of the genus Vesiculovirus in the family Rhabdoviridae. These methods also revealed that SVCV is not
42 104
related to fish rhabdoviruses of the genus Novirhabdovirus. In vitro replication of SVCV takes place in
43 the cytoplasm of cultured cells of fish, bird and mammalian origin at temperatures of 4 to 31°C, with an 105
44 optimum of about 20°C. Spring viremia of carp can be diagnosed by clinical signs, isolation of virus in cell 106
45 culture and molecular methods. Antibodies directed against SVCV react with the homologous virus in 107
46 serum neutralization, immunofluorescence, immunoperoxidase, or enzyme-linked immunosorbent 108
47 assays, but they cross-react to various degrees with the pike fry rhabdovirus (PFR), suggesting the 2 109
48 viruses are closely related. However, SVCV and PFR can be distinguished by certain serological 110
49 tests and molecular methods such as the ribonuclease protection assay. 111
50 112
51 KEY WORDS: Rhabdovirus · Spring viremia of carp virus · SVCV · Disease · Cyprinids · Carp · Fish disease 113
52 Resale or republication not permitted without written consent of the publisher 114
53 115
54 116
55 INTRODUCTION ously known as infectious dropsy, rubella, infectious 117
56 ascites, hemorrhagic septicemia, and red contagious 118
57 Dating back possibly to the Middle Ages, common disease (Hofer 1904, Schäperclaus 1930, Toma$ec et 119
58 carp Cyprinus carpio in European pond culture have al. 1964, Bauer & Faktorovich 1969). These diseases 120
59 been plagued by a complex of infectious diseases vari- proved to be of great economical importance, causing 121
60 122
61 © Inter-Research 2002 · www.int-res.com *Corresponding author. 123
62 E-mail: sandra.essbauer@micro.vetmed.uni-muenchen.de 124
125 2 Dis Aquat Org ■ ■ 187
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129 serious losses in carp pond fisheries of the central vent (Fig. 1A). Internal signs are peritonitis, ascites, 191
130 and eastern parts of Europe (Fijan 1972, 1999). The catarrhal and hemorrhagic enteritis, edematous vis- 192
131 assumed causes (nutrition, environment, parasites, cera and petechial hemorrhages of the internal wall of 193
132 bacteria, viruses) for the acute and chronic forms of the the swimbladder and in skeletal muscle (Fijan et al. 194
133 epizootics remained controversial for a long time. 1971, Ahne & Wolf 1977, Negele 1977). 195
134 However, a viral etiology for the acute form of infec- A rhabdovirus isolated from penaeid shrimps in 196
135 tious dropsy became evident when a cytopathic agent Hawaii (Lu et al. 1991) had a G gene that exhibited 197
136 was isolated (Osadchaya 1964, Toma$ec et al. 1964), over 99% identity to the G nucleotide sequence of 198
137 and River’s postulates were fulfilled using virus iso- SVCV (Johnson et al. 1999). The agent caused histo- 199
138 lated from affected carp (Fijan et al. 1971). In order to logical changes and significant mortality in penaeid 200
139 distinguish the viral disease from other aetiological shrimps (Lu & Loh 1994). The SVCV-like virus isolated 201
140 entities within the infectious dropsy complex, the dis- from penaeid shrimp in Hawaii was reported to cause 202
141 ease was renamed ‘spring viremia of carp’ (SVC), and histological changes and mortalities in Penaeus 203
142 the causative virus was termed ‘spring viremia of carp stylirostris through experimental water-born infection, 204
143 virus’ (SVCV) or Rhabdovirus carpio (Fijan et al. 1971). oral feeding and intramuscular injection of the virus 205
144 The infectious disease of carp termed swim-bladder (Lu et al. 1991). 206
145 inflammation (SBI) was also recognized in Europe 207
146 (Arshaniza et al. 1968, Otte 1972). A rhabdovirus iso- 208
147 lated from carp exhibiting the acute form of SBI (Ahne Properties of SVCV 209
148 1973, Bachmann & Ahne 1973, 1974) was found to be 210
149 identical to R. carpio (de Kinkelin & Le Berre 1974, Hill SVCV exhibits the typical bullet-shaped morpho- 211
150 et al. 1975). Later, it was shown that the chronic dis- logy of a vertebrate rhabdovirus (Wunner et al. 1995) 212
151 ease termed ‘carp erythrodermatitis’ was caused by a (Fig. 1B). The virion possesses an inner nucleocapsid 213
152 bacterium of the genus Aeromonas (Bootsma et al. with helical symmetry measuring about 50 nm in dia- 214
153 1977). For more complete reviews of the history of meter. The virion measures approximately 80 to 180 nm 215
154 these conditions, see Fijan (1999) or Wolf (1988). in length and 60 to 90 nm in diameter. Truncated par- 216
155 In Europe, SVCV mainly affects the common carp ticles, about 2/3 of the length of infective virions, 217
156 Cyprinus carpio, but several other species of fresh represent defective, noninfectious virions (Fijan et al. 218
157 water fishes can be infected (Fijan 1972, 1988, 1999, 1971, Bachmann & Ahne 1973, Hill et al. 1975, Bishop 219
158 Wolf 1988). In carp, the most common external signs & Smith 1977). SVCV has a buoyant density in CsCl 220
159 of SVC are hemorrhages of the skin, exopthalmia, of 1.195 to 1.200 g cm– 3 (Bachmann & Ahne 1973). 221
160 abdominal distension, and an inflamed or edematous Virus infectivity is destroyed at pH 3 and 12, by lipid 222
161 223
162 224
163 A B 225
164 Fig. 1. Cyprinus carpio. 226
165 (A) SVCV (spring viremia of 227
166 carp virus)-infected com- 228
167 mon carp showing petechial 229
hemorrhages in the mus-
168 230
cles, swim bladder, enlarged
169 spleen, edematous organs 231
170 and pale gills with hemor- 232
171 rhages. (B) Electron micro- 233
graph of SVCV in infected
172 234
fathead minnow (FHM) cells
173 C D showing bullet-shaped parti- 235
174 cles. (C) Cytopathogenic ef- 236
175 fect (CPE) of SVCV in FHM 237
176 cells, 72 h after infection, 238
20°C. The CPE is character-
177 239
ized by rounding of cells and
178 focal lysis of the monolayer 240
179 (magnification: 200 ×). (D) 241
180 Detection of SVCV antigen 242
in FHM cells, 24 h post injec-
181 243
tion (p.i.) using indirect im-
182 munofluorescence (magnifi- 244
183 cation: 90 ×) 245
184 246
185 247
186 248
249 Ahne et al.: Spring viremia of carp 3 311
250 312
251 313
252 314
253 Table 1. Features of the 5 genes of the spring viremia of carp virus (SVCV) the most abundant virion protein and 315
254 genome. N: nucleocapsid; P: phosphoprotein; M: matrix protein; G: glycopro- interacts with the viral RNA to form 316
255 tein; L: polymerase. nt: nucleotides; aa: amino acids; mw: molecular weight; 317
the helical structure of the nucleo-
pI: isoelectric point. The last 3 columns refer to the protein predicted by transla-
256 capsid, and also has an important role 318
tion of the open reading frames (ORFs) in the nucleotide sequence of the SVCV
257 genome. Thus, the protein molecular weight is for the predicted product with no in modulating transcription. About 1/3 319
258 post-translational modification such as phosphorylation or glycosylation of the N protein is associated with 320
259 viral RNA (Sokol & Koprowski 1975). 321
260 The phosphoprotein (P protein) is a 322
Gene Total gene Upstream Downstream ORF Protein Protein Protein
261 length (nt) UTR (nt) UTR (nt) (nt) (aa) (mw) (pI) component of the rhabdovirus nucleo- 323
262 capsid that, in association with the 324
263 N 1335 10 68 1254 418 47000 5.57 L and N proteins, is required for 325
264 P 967 10 27 927 309 35622 4.46 326
transcription (Wunner et al. 1975,
M 716 10 34 669 223 25627 8.61
265 G 1588 10 48 1527 509 57338 5.54 Roy 1981). Like other vesiculoviruses, 327
266 L 6325 10 (trailer) 6285 2095 238244 8.63 SVCV has one membrane protein 328
267 (M protein). This basic protein forms 329
268 the bullet-shaped structure of the 330
269 virion and links the nucleocapsid with 331
270 solvents, and by heat (56°C). The virus is inactivated the cytoplasmic domains of the G protein embedded 332
271 within 10 min by formalin (3%), chlorine (500 ppm), in the lipid-containing viral envelope (Walker et al. 333
272 iodine (0.01%), NaOH (2%), UV (254 nm) and gamma 2000) (Fig. 2). 334
273 irradiation (103 krads). Freeze-thaw cycles partially 335
274 inactivate the virus. During lyophilization, addition of 336
275 2 to 10% fetal calf serum protects the virus infectivity Genomic organization of SVCV 337
276 (de Kinkelin & Le Berre 1974, Ahne 1976, 1982). 338
277 The virus is presently classified as a tentative mem- The SVCV virion contains 1 molecule of linear, 339
278 ber of the genus Vesiculovirus of the family Rhabdo- negative-sense, single-stranded RNA that sediments 340
279 viridae (Walker et al. 2000). As with other members of in 5 to 20% sucrose gradients at 38 to 40 S (Hill et al. 341
280 the genus, the virion of SVCV contains 5 structural 1975, Lenoir & de Kinkelin 1975, Clerx & Horzinek 342
281 proteins (now termed N, P, M, G, L), the sizes of which 1978, Roy & Clewley 1978, Wu et al. 1987). Character- 343
282 were initially estimated by mobility in SDS-PAGE ization of this SVCV genomic RNA began in 1984 with 344
283 (sodium dodecyl sulphate polyacrylamide gel elec- publications of the M gene sequence and a 70 nucleo- 345
284 trophoresis) gels (Lenoir 1973, Lenoir & de 346
285 Kinkelin 1975, Clerx & Horzinek 1978, Roy 347
286 et al. 1984). Recently, the availability of 348
287 sequence data for the entire SVCV genome A
a) SVCV Virion L Polymerase 349
288 allows more accurate predictions of the 350
289 molecular weights of the structural proteins G Glycoprotein 351
290 (ignoring post-translational modifications) 352
N Nucleoprotein
291 as well as other features of these molecules 353
292 (Table 1). The RNA-dependent RNA poly- P Phosphoprotein 354
293 merase (L protein) functions in transcription 355
M Matrix protein
294 and replication of the virus with an optimal 356
Envelope
295 temperature for activity between 20 and 357
296 25°C (Roy & Clewley 1978). Virus transcrip- 358
297 tion and replication is achieved as the L pro- 359
298 tein interacts with the P and N proteins of B
b) SVCV Genome RNA (11,019 nt) 360
299 the nucleocapsid. The glycoprotein (G pro- 3’ N P M G L 5’ 361
300 tein) of SVCV forms trimeric peplomers or 362
301 spikes on the virus surface that bind to cel- 363
302 lular receptors and induce viral endocytosis. c) 364
C conserved gene junction and upstream untranslated region:
303 The surface glycoprotein acts as the most 365
304 important viral antigen that determines the ...TATG(A) CTAACAGASATCATG... 366
7
305 serological properties of rhabdoviruses (Hill 367
306 et al. 1975, Bishop & Smith 1977, Jørgensen Fig. 2. Spring viremia of carp virus (SVCV). (A) Virion structure, (B) ge- 368
307 et al. 1989). The nucleoprotein (N protein) is nomic organization, and (C) gene junction sequences of SVCV 369
308 370
309 371
310 372
373 4 Dis Aquat Org ■ ■ 435
374 436
375 437
376 438
377 tide sequence at the 3’ genome terminus (Kiuchi & Roy a tetranucleotide (CTAT). These regulatory signals 439
378 1984, Roy et al. 1984). This line of investigation then and intergenic regions are highly conserved between 440
379 lays apparently dormant until publication of the partial SVCV and all vesiculoviruses, and they differ from 441
380 L gene, complete G gene, and gene junction se- the gene junctions of viruses in other rhabdovirus 442
381 quences in 1995 and 1996 (Bjorklund et al. 1995, 1996). genera such as lyssaviruses or novirhabdoviruses. 443
382 In 2001, 2 complete genomic RNA sequences of an However, unlike other vesiculoviruses that have been 444
383 American type culture collection (ATCC) reference characterized to date, the untranslated regions up- 445
384 strain of SVCV (VR-1390) were submitted to Gen- stream of all 5 SVCV genes are completely uniform, 446
385 Bank by independent laboratories (GenBank accession consisting of exactly 10 nucleotides preceding the 447
386 U18101 by Bjorklund et al. 1996 and GenBank acces- ATG-codon signaling the start of translation (Table 1). 448
387 sion AJ318079 by Hoffman et al.). With this data now The sequence of this 10-base region begins with the 449
388 available, the genetic organization of SVCV can be recognized vesiculovirus pentanucleotide transcrip- 450
389 definitively described. The genomic RNA is 11019 tion start signal and is highly conserved between the 451
390 bases in length. It contains 5 major open reading gene junctions: 4 of the SVCV genes (N, M, G, L) 452
391 frames (ORFs), encoding predicted proteins homolo- begin with the sequence AACAGACATC, and only 453
392 gous to the 5 major structural proteins found in all the P gene has a single nucleotide difference making 454
393 rhabdoviruses. The genomic order is 3’-NPMGL-5’, as it AACAGAGATC. Although the length of the up- 455
394 shown in Fig. 2B. This is identical to the genetic orga- stream untranslated regions is variable among other 456
395 nization of species in the genera Vesiculovirus and vesiculovirus genes, they begin with a consensus 457
396 Lyssavirus. This confirmed gene order for SVCV clari- sequence AACAGNNATC (Luis L. Rodriguez, USDA 458
397 fies any confusion resulting from an early publication Plum Island Animal Disease Center, pers. comm.), 459
398 that suggested an alternative organization (Wu et al. which matches the conserved SVCV sequence at 8 460
399 1987). The SVCV genome does not have a non-virion out of 10 sites. This identity at the 3’ termini of the 461
400 (NV) gene between the G and L genes as is found genes provides confirmation for an early comparison 462
401 in fish rhabdoviruses of the genus Novirhabdovirus of transcript termini of SVCV and vesicular stomatitis 463
402 (Kurath et al. 1997, Walker et al. 2000). Similarly, it Indiana virus (VSIV) that suggested conservation of 464
403 does not have a large non-coding region between the the rhabdoviral genome sequences specifying the 5’ 465
404 G and L gene ORFs, as is found in lyssavirus genomes. ends of transcript RNAs (Gupta et al. 1979). The tran- 466
405 Thus, the overall genome structure of SVCV is most scription stop/polyadenylation signal for the SVCV L 467
406 similar to that of the vesiculovirus genus. gene is followed by a 46 base trailer that forms the 468
407 The SVCV genome has a 59 base putative leader 5’ terminus of the genome. 469
408 region at the 3’ terminus followed by a consensus Sequence similarities between SVCV and vesicular 470
409 start signal (AACAG; mRNA-sense) for transcription stomatitis viruses were first reported in 1984 using 471
410 of the N gene. Analysis of the SVCV N-gene mRNA complete M gene and 3’ terminal genome sequences 472
411 by 5’ RACE has confirmed that transcription starts at (Kiuchi & Roy 1984, Roy et al. 1984). The 3’-terminal 473
412 this AACAG signal (W. Batts & G. Kurath unpubl.). 20 nucleotides were found to be nearly identical to the 474
413 The features of the 5 genes and their protein products corresponding sequences of both VSIV and VSNJV. 475
414 are summarized in Table 1. Other than the 5 major Later, comparisons of the first 1780 amino acids of the 476
415 ORFs, only one is longer than 50 amino acids in the predicted SVCV L protein with partial L proteins of 477
416 standard rhabdovirus orientation (53 amino acids, other rhabdoviruses revealed closest homology with 478
417 within the M gene), and there are several in the the VSIV polymerase, although significant homology 479
418 reverse orientation, all less than 140 amino acids. It is was also found with the polymerase of rabies virus 480
419 not known whether any of these small ORFs are (Bjorklund et al. 1995). The homologies involved sev- 481
420 expressed, and there is no indication of an over- eral highly conserved domains separated by variable 482
421 lapping reading frame within the P gene encoding a regions. The conserved domains most likely represent 483
422 small basic protein, as has been reported for vesicular previously identified RNA polymerase functional do- 484
423 stomatitis New Jersey virus (VSNJV) (Spiropoulou & mains such as those for RNA binding, RNA template 485
424 Nichol 1993). recognition, phosphodiester bond formation, and ribo- 486
425 As described by Bjorklund et al. (1996), the 4 nucleotide triphosphate binding (Poch et al. 1990). 487
426 SVCV gene junctions are strictly conserved, with a There was very little similarity between the L proteins 488
427 TATG(A)7 transcription stop/polyadenylation signal at of SVCV and the novirhabdovirus, infectious hemato- 489
428 the end of each gene, and an AACAG transcription poietic necrosis virus. The SVCV G gene encodes a 490
429 start signal for the following gene. The untranscribed 509 amino acid protein with a predicted structure 491
430 intergenic regions of SVCV are all dinucleotides (CT), similar to other rhabdovirus glycoproteins (Bjorklund 492
431 with the exception of the G-L gene junction which has et al. 1996). It contains 2 major hydrophobic domains 493
432 494
433 495
434 496
497 Ahne et al.: Spring viremia of carp 5 559
498 560
499 561
500 562
501 including the signal peptide and transmembrane do- relationship between SVCV and accepted members of 563
502 main. The complete SVCV G protein was most closely the Vesiculovirus genus. This is supported by signifi- 564
503 related to members of the vesiculovirus genus, show- cant bootstrap values, providing a high level of con- 565
504 ing 31.2 to 33.2% amino acid identity (51.8 to 53.3% fidence in this relationship. The 7th report of the 566
505 amino acid similarity) with VSIV, VSNJV, and chandi- International Committee for Virus Taxonomy lists 567
506 pura virus (CHPV). In contrast, there was only 19.4 to SVCV as a tentative member of the vesiculovirus 568
507 24.3% amino acid identity (40.5 to 47.2% amino acid genus (Walker et al. 2000). In the future, data includ- 569
508 similarity) with G proteins of 11 rhabdoviruses from ing thorough phylogenetic analyses will be used in 570
509 other genera. support of a proposal that SVCV belongs within this 571
510 Phylogenetic analyses (Fig. 3) using the full-length genus as a full member, as has been suggested previ- 572
511 amino acid sequences of the N, P, M, and G proteins, ously (Bjorklund et al. 1996). 573
512 as well as partial sequences of the L proteins, con- Within the species SVCV, the genetic diversity 574
513 firms the close relationship between SVCV and the among SVCV strains has been demonstrated by bio- 575
514 established members of the Vesiculovirus genus logical and serological analyses (Jørgensen et al. 1989) 576
515 (Bjorklund et al. 1996, Johansson 2001). Examples of and by ribonuclease protection assays (Ahne et al. 577
516 such phylogenetic trees are shown in Fig. 3 for full- 1998). Although there are not yet any published re- 578
517 length G and N protein sequences. Despite minor dif- ports characterizing sequence diversity between strains, 579
518 ferences in topology between the trees generated by this is an area of active investigation in several labora- 580
519 the different genes, both indicate a close phylogenetic tories. 581
520 582
521 583
522 584
523 A) N Proteins B) G Proteins 585
524 100 VHSV-0771 VSNJV 586
100
525 83 VHSV-Mak 100 VSIV 587
V
526 100 SHRV N 100 CHPV 588
527 100 IHNV 100 SVCV * 589
528 HIRRV SIGMA 590
529 100 VSNJV VHSV-Mak 591
100
530 71 VSIV 100 592
V 100 VHSV-0771
531 100 SVCV * 593
100 SHRV N
532 99 CHPV 100 HIRRV 594
98
533 100 ARV IHNV 595
E
534 BEFV 100 MOKV 596
100
L
535 100 MOKV RABPV 597
L
536 RABPV ARV 598
100 E
537 SIGMA BEFV 599
538 LNYV C LNYV C 600
539 SYNV Nu SYNV Nu 601
540 0.1 changes 0.1 changes 602
541 603
542 Fig. 3. Phylogenetic relationships of (A) nucleocapsid and (B) glycoprotein gene sequences of spring viremia of carp virus (SVCV) 604
543 and 15 other rhabdoviruses. The viral species and the GenBank sources for their N and G amino acid sequences were: SVCV 605
(U18101); vesicular stomatitis New Jersey virus (VSNJV, K02379, J02433); vesicular stomatitis Indiana virus (VSIV, J02428);
544 606
Chandipura virus (CHPV, M16608, J04350); viral hemorrhagic septicemia virus, strain 0771 (VHSV-0771, AJ233396); viral hem-
545 orrhagic septicemia virus, strain Makah (VHSV-Mak, X59241, U28747); infectious hematopoietic necrosis virus (IHNV, L40883); 607
546 Hirame rhabdovirus (HIRRV, AF104985, U24073); snakehead rhabdovirus (SHRV, AF147498); Rabies virus, Pasteur vaccine 608
547 strain (RABPV, M13215); Mokola virus (MOKV, Y09762, S59447); Adelaide River virus (ARV, U10363, L09207); bovine ephemeral 609
548 fever virus (BEFV, AF234533); Sigma virus (SIGMA, X91062); lettuce necrotic yellows virus (LNYV, L30103, AJ251533); and 610
sonchus yellow net virus (SYNV, L32603). Sequences were aligned using Clustal X (Thompson et al. 1997) and analyzed with
549 PAUP* 4.0 (Swofford 2000) neighbor-joining (shown) and parsimony phylogeny programs (not shown). Sequences from the plant 611
550 rhabdovirus SYNV were used to outgroup the analyses. Each tree represents the consensus of 1000 bootstrapped replicates of the 612
551 data, with bootstrap confidence values along the branches to indicate the percent of 1000 trees that contained exactly the group- 613
552 ing shown to the right of the value. Branches with bootstrap values less than 70 were collapsed to avoid suggesting uncertain re- 614
lationships. The viral species included represent the 6 accepted rhabdovirus genera and one unassigned species (Sigma virus).
553 615
Genera are indicated by brackets labeled as follows: V, Vesiculovirus; N, Novirhabdovirus; L, Lyssavirus; E, Ephemerovirus; C,
554 Cytorhabdovirus; Nu, Nucleorhabdovirus. Note that the position of SVCV (in bold and asterisked) relative to the 3 accepted 616
555 members of the Vesiculovirus genus (VSNJV, VSIV, and CHPV) suggests that SVCV should be included in this genus 617
556 618
557 619
558 620
621 6 Dis Aquat Org ■ ■ 683
622 684
623 685
624 686
625 In vitro multiplication of SVCV 1975, Fijan et al. 1977a,b, Baudouy 1978, Ahne 1980, 687
626 Baudouy et al. 1980a,b, Fijan & Mata$in 1980, Fijan 688
627 SVCV replicates in cell cultures originated from fish, 1988). Rhabdovirus-neutralizing antibodies are directed 689
628 birds and mammals between 4 and 31°C (Ahne 1973, against the surface glycoprotein of the virus (Kelly 690
629 Clark & Soriano 1974). Cytopathic effects (CPE), like et al. 1972). Induction of humoral antibodies against 691
630 margination of nuclear chromatin followed by rounding SVCV in carp is influenced by the age and condition of 692
631 up and lysis of cells (Fig. 1C), appear at varying times of carp, by the route of infection and, most importantly, 693
632 infection depending on the temperature and cell lines by the temperature of water (Fig. 4). In carp which 694
633 used. The optimal temperature of viral replication in were infected by waterborne exposure to low doses of 695
634 vitro is between 20 and 25°C. The best cell systems for SVCV and kept at 20°C, SVCV neutralizing antibodies 696
635 multiplication of SVCV are those derived from cyprinid appeared 7 d after infection. In contrast, carp infected 697
636 fishes (e.g. primary cell cultures or established cell in the same way, but kept at 13°C, showed first de- 698
637 lines) such as the epithelioma papillosum cyprini (EPC) tectable antibodies 7 wk after infection. At 13°C, carp 699
638 cell line (Fijan et al. 1983), the fathead minnow (FHM) developed a subclinical infection with presence of 700
639 cell line (ATCC CCL-42, Gravell & Malsberger 1965), virus in the blood of carp for about 10 wk. Neutralizing 701
640 and carp leukocyte cultures (CLC, European Collection antibodies which appeared 8 to 10 wk after infection 702
641 of Cell Cultures 95070628, Faisal & Ahne 1990). Using lead to a rapid decline of the amount of virus in the 703
642 these cell lines, infectivity titers of about 108 TCID50 blood (Ahne 1979, 1980, 1986). The biological proper- 704
643 ml–1 were obtained regularly. Actinomycin D does not ties of the SVCV neutralizing antibodies have not been 705
644 affect SVCV replication (de Kinkelin & Le Berre 1974). studied in detail, but probably resemble the tetrameric 706
645 The virus forms well-defined plaques in permissive antibody typically found in teleost fishes (Fijan et al. 707
646 cells 72 h after infection. A single step growth curve 1977a, Kaattari & Piganelli 1996). 708
647 carried out in FHM cells at 20°C showed that the first Cross-neutralization studies using the fish rhabdo- 709
648 progeny virus is synthesized 4 to 6 h after infection. viruses, infectious hematopoietic necrosis virus (IHNV), 710
649 Both cell-associated and cell-free virus reach peak viral hemorrhagic septicemia virus (VHSV), hirame 711
650 titers between 10 and 22 h after infection. One growth rhabdovirus (HIRRV) and their respective antisera, 712
651 cycle of the virus lasts 8 to 10 h at 20°C (Ahne 1973, revealed no antigenic relationships between SVCV 713
652 Bachmann & Ahne 1974). SVCV also replicates in and these members of the Novirhabdovirus genus of 714
653 chicken embryo fibroblasts, in mammalian cells such the Rhabdoviridae (de Kinkelin & Le Berre 1974, de 715
654 as BHK 21 (hamster), fetal calf kidney (cattle), HEP-2 Kinkelin et al. 1974, Hill et al. 1975). However, sero- 716
655 (human), MDCK (canine), SK (porcine), Vero (monkey) logical examination of 22 rhabdovirus isolates from 717
656 and in several reptilian cell lines, provided that incuba- 718
657 tion temperatures between 20 and 22°C are used (Ahne 719
658 1973, Bachmann & Ahne 1974, Clark & Soriano 1974). It 720
659 has been demonstrated that apoptosis is involved in 721
660 death of EPC cells caused by SVCV (Bjorklund et al. 722
661 1997). Morphological changes, like reduction of cell 723
662 volume, blebbing, and formation of apoptotic bodies, 724
663 appear 36 h after infection of cells. About 40 h later, 725
664 more than 70% of the SVCV-infected EPC cells were 726
665 found to be apoptotic. It is likely that the programmed 727
666 cell death of the SVCV-infected cells depends on 728
667 replication and production of progeny virus. Apoptosis 729
668 could be inhibited by the endogenous acid cysteine 730
669 proteinase inhibitor, cystatin A, which has been puri- 731
670 fied from human palatine tonsils (Bjorklund et al. 1997). 732
671 Experiments showed that SVCV-induced apoptosis can 733
672 also be inhibited by z-VAD-fmk, an inhibitor of caspase 734
673 1, 3, 4 and 7 (Taehtinen et al. 1999). 735
674 Fig. 4. Influence of water temperature on SVCV (spring 736
675 viremia of carp virus)-infection of carp Cyprinus carpio lead- 737
676 Antigenic properties of SVCV ing to virus multiplication (10 to 12°C) or to production of anti- 738
bodies (20 to 22°C). SPF carp (25 to 30 g) were intraperi-
677 739
toneally infected with 103 TCID50 of SVCV. Infected fish kept
678 Several studies showed that carp develop a humoral at 10 to 12°C and 20 to 22°C were analyzed for values of 740
679 immune response to SVCV (Sulimanović 1973, Kölbl virus, antibodies and death (Ahne 1980) 741
680 742
681 743
682 744
745 Ahne et al.: Spring viremia of carp 7 807
746 808
747 809
748 810
749 Cyprinidae, Esocidae and Siluridae genera indicates The range of alterations of the liver went from perivas- 811
750 close relationship between SVCV and the vesiculo- culitis showing lymphocytes and histiocytes infiltra- 812
751 virus-like fish rhabdovirus, pike fry rhabdovirus (PFR, tion, to panvasculitis with a high grade of edematiza- 813
752 Jørgensen et al. 1989). These authors showed that tion and a concurrent loss of structure of blood vessel 814
753 SVCV and PFR cross-reacted by indirect immuno- walls. The blood vessels appeared to be fully necrotic 815
754 fluorescence (Fig. 1D) using polyclonal antisera and in the final stage of the disease. The liver parenchyma 816
755 shared common antigenic determinants on the G, N showed multifocal necroses, adipose degeneration and 817
756 and M proteins as revealed by immunoblotting. The 2 hyperemia. In the pancreas, non-purulent inflamma- 818
757 viruses could, however, be differentiated by neutral- tion and multifocal necrobiosis were usually seen. In 819
758 ization assays using certain polyclonal antisera in the the parietal and visceral serosa of the peritoneum, 820
759 absence of complement. Consequently, it was sug- peritonitis was predominant. Lymph vessels were 821
760 gested that SVCV and PFR should be considered as extremely dilated and filled up with detritus, macro- 822
761 representatives of 2 serotypes of 1 virus species (Jør- phages, and lymphocytes. In the intestine, perivascular 823
762 gensen et al. 1989). The 2 viruses could also be 824
763 differentiated by a ribonuclease protection assay 825
764 using a probe made from transcripts of the G gene PFU ml–1 (g tissue) 826
765 of SVCV (Ahne et al. 1998), indicating that 827
766 genetic differences exist between the 2 viruses. 828
767 Resolution of the biological and genetic relation- 829
768 ships between isolates of SVCV and PFR is im- 107 pools of 830
kidney,
769 portant because isolation of SVCV from fish is liver, 831
Virus titers in blood, gills and organs of freshly killed carp

spleen
770 notifiable to the Office International des Epi- and gut 832
771 zooties (OIE) while PFR is not. 833
772 834
106
773 835
774 Experimental infection of carp with SVCV 836
775 837
776 The systemic character of SVCV infection has gills 838
777 been demonstrated by experimental infection of 839
105
778 specific pathogen free (SPF) carp (Ahne 1977, virus amount 840
in the water
779 1978). Following waterborne infection of carp at 841
gills blood
780 13°C, the virus was initially detected in the gills, 842
virus
781 suggesting they were the first targeted organ. amount 843
782 Following an eclipse-like period of 4 d, SVCV was in feces 844
104
783 spread via blood to the internal organs of infected 845
virus in gills
784 fish. Eleven days after infection, SVCV was after 2 h bath 846
785 excreted from the alimentary tract in the feces 847
primary virus-
786 and mucous casts. First mortality due to SVCV multiplication 848
103 at the side of entry
787 infection appeared 20 d after infection. The incu- 849
blood streams

20% mortality
virus in feces

788 850
virus enters

bation period of the water borne SVCV infection

secondary

first death

alimentary tract
primary
viremia

viremia

789 was found to be 7 d under the experimental con- 851

virus cannot be
790 ditions used (Fig. 5). First clinical signs such as detected in gills, 852
791 blood, organs 853
edematous viscera, peritonitis, enteritis and hem- and skin
792 102 854
orrhages in different organs could be recognized 1 2 3 4 5 6 7 8 9 10 11 12 13 20 30 days after
793 8 to 11 d after infection. The virus persisted in infection 855
794 infected carp for more than 10 wk and those fish Incubation period Clinical disease with 856
795 became SVCV carriers (Ahne 1977, 1978, 1980). pathological lesions 857
796 858
Fig. 5. Spread of spring viremia of carp virus (SVCV) through the
797 859
body of carp Cyprinus carpio after waterborne infection for 2 h at
798 Histopathology of SVC 13°C. SPF (specific pathogen free) carp (25 to 30 g) were intraperi- 860
799 toneally infected with 104.4 plaque-forming units (PFU) ml–1 of SVCV 861
800 Precise reports on histological changes of by waterbath at 13°C. Two hours after infection, SVCV was detected 862
only in the gills. After a 4 d eclipse period, progeny viruses were
801 SVCV-infected carp are rare. The following histo- 863
dessimated to internal organs via the bloodstream. Two viremia
802 logical findings have been obtained in experi- stages were recognized 6 and 10 d after infection. The incubation 864
803 mentally infected carp fingerlings (Negele 1977). period lasted 7 d at 13°C (Ahne 1978) 865
804 866
805 867
806 868
869 8 Dis Aquat Org ■ ■ 931
870 932
871 933
872 934
873 inflammation and desquamation of epithelium with a neutralizing antibodies appear about 1 wk after infec- 935
874 subsequent atrophy of the villi have been often recog- tion at 20°C. Immune fish undergo a solid specific re- 936
875 nized. The spleen was hyperemic and showed a con- sistance against reinfection. The persistence of SVCV 937
876 siderable hyperplasia of the reticuloendothelium. antibodies may vary individually from a few to several 938
877 Siderocytes and cells with increased lipofuscin storage months (Fijan et al. 1977a, Baudouy 1978, Ahne 1979, 939
878 were present. The tubuli of the kidney were clogged Fijan 1988). 940
879 by the tube casts, and vacuolation and hyaline degen- 941
880 eration were present. The lamina epithelia of the swim 942
881 bladder changed into a discontinuous multilayer and Epizootiology of SVC 943
882 in the submucosa dilated vessels and hemorrhages 944
883 were evident. The heart showed discontinuous myode- The common carp Cyprinus carpio is the main host of 945
884 generation and inflammatory alterations of the peri- SVCV (Fijan 1999). The virus affects carp at all ages 946
885 cardium. but victims are mostly young fish. Natural outbreaks 947
886 of SVC were also found in koi carp C. carpio, crucian 948
887 carp Carassius carassius, big head Hypophthalmich- 949
888 Natural outbreaks of SVC thys molitrix) silver carp Aristichthys nobilis and sheat- 950
889 fish Silurus glanis (Fijan et al. 1984, Jørgensen et al. 951
890 SVC is reported to be present in several European 1989, Shchelkunov & Shchelkunova 1989, Ahne et al. 952
891 countries (Austria, Bulgaria, France, Germany, Great 1998). Grass carp Ctenopharyngodon idella, Guppies 953
892 Britain, Hungary, Italy, Spain, as well as in parts of the Lebistes reticulatus, northern pike Esox lucius, pump- 954
893 former Czechoslovakia, Soviet Union and Yugoslavia; kinseed Lepomis gibbosus and roach Rutilus rutilus 955
894 Toma$ec et al. 1964, Fijan et al. 1971, Ghittino et al. proved to be susceptible to SVCV by experimental 956
895 1971, Ahne 1973, Baudouy 1975, Kölbl 1975, Tesar<ik infection (Ahne 1973, 1985b, Fijan 1988, Haenen & 957
896 et al. 1977, Bucke & Finlay 1979, Roudikov 1980, Davidse 1993). SVCV can persist in asymptotic brood- 958
897 Békési & Csontos 1985, Shchelkunov & Shchelkunova stock carp (Békési & Csontos 1985). Reservoir hosts 959
898 1989, Marcotegui et al. 1992). In those countries, include sick fish and carp which survive epizootics, as 960
899 SVCV has economic significance in carp pond cul- well as other cultured or wild fish that may serve as 961
900 tures. Mortality rates of young carp due to SVC can carriers of SVCV. Possible biological vectors include 962
901 reach up to 70% during spring time outbreaks, but the the carp louse Argulus foliaceus and leeches Piscicola 963
902 yearly losses of older fish are usually below 30%. Out- geometra, as well as fish-eating birds such as herons 964
903 breaks of SVC depend on the temperature of the (Pfeil-Putzien 1977, Ahne 1985a, Peters & Neukirch 965
904 water, age and condition of fish, population density, 1986). It was demonstrated that the fruit fly Drosophila 966
905 and stress factors. SVCV-infected carp in ponds tend to melanogaster multiplied SVCV under experimental 967
906 concentrate around the water inlet. Reactions to sen- conditions (Bussereau et al. 1975). While the role of 968
907 sory stimulation, swimming speed, and the respiration strictly terrestrial insects in the transmission of SVCV 969
908 rate are slowed down progressively; lethargy, resting is unlikely, aquatic arthropods may be able to function 970
909 and leaning mark the terminal stage of disease. Exter- as SVCV vectors. The SVCV-like agent isolated from 971
910 nal signs of SVC under natural conditions are dark- shrimp in Hawaii replicated and caused histological 972
911 ening of the skin, distended abdomen, exophthalmia, changes in such crustaceans (Lu & Loh 1994). 973
912 petechial hemorrhages in the skin, gills and eyes, in- Generally speaking, transmission of SVCV is hori- 974
913 flamed and edematous vent, and pale gills. Internally, zontal. Excretion of SVCV via feces and urine from 975
914 hemorrhages in the swim bladder, edematous organs, infected fish has been demonstrated. The excreted 976
915 ascites and catarrhal enteritis are usually seen. Sec- virus could stay infective in the water for more than 977
916 ondary and concomitant bacterial and/or parasitic 4 wk and in mud for about 6 wk at 4 to 10°C. An addi- 978
917 infections can affect the mortality rate and symptoms tional source of infection may be contaminated equip- 979
918 (Fijan 1972). Patterns of the disease are influenced by ment (Ahne 1977, 1979, 1982). Several studies suggest 980
919 the mentioned environmental factors as well as by that vertical transmission of SVCV is not an important 981
920 unspecific (interferon) or specific immunity (humoral source of infection (Ahne 1979, Békési & Csontos 1985, 982
921 antibodies, cellular immunity). Clinical disease domi- Fijan 1988, 1999). 983
922 nates at water temperatures between 5 and 10°C, 984
923 where the host immune response is usually suppressed 985
924 or delayed. However, the lethal outcome is faster at Diagnosis of SVC 986
925 temperatures between 10 and 17°C, and fry can be 987
926 affected at temperatures as high as 22 to 23°C. SVCV- SVC can be expected to cause mortalities in carp 988
927 infected carp develop interferon within 24 h, and virus aquaculture at temperatures below 20°C, especially in 989
928 990
929 991
930 992
993 Ahne et al.: Spring viremia of carp 9 1055
994 1056
995 1057
996 1058
997 the spring. The behavior and external signs in easily 1994). As already described above, SVCV shares anti- 1059
998 caught moribund fish exhibiting hemorrhages in skin, genic determinants with PFR (Jørgensen et al. 1989, 1060
999 pale gills, ascites, etc., warrant necropsy and further Way 1991). SVCV and PFR cannot be reliably distin- 1061
1000 sampling of fish for virological examination. Presump- guished by certain serological approaches, especially 1062
1001 tive diagnosis is based on the presence of enteritis, IF and ELISA. Both rhabdoviruses are listed as tenta- 1063
1002 peritonitis, edema, petechial bleedings in the swim tive species in the genus Vesiculovirus of the Rhab- 1064
1003 bladder, muscles and other organs. Lack of some signs doviridae (Walker et al. 2000), but PFR is, in contrast to 1065
1004 or their slight pronouncement do not exclude the need SVC, not an OIE notifiable agent. Therefore, proper 1066
1005 for laboratory examination. Variations and differences methods should be used to distinguish between iso- 1067
1006 of disease signs in a population should be encom- lates of SVCV and PFR. A ribonuclease protection 1068
1007 passed by a selection of separate samples, and all assay (RPA) was developed that can distinguish be- 1069
1008 affected species should be sampled. Whole fishes near tween SVCV and PFR (Ahne et al. 1998). Probes for 1070
1009 death or freshly dead specimens should be chilled and the full-length G gene of SVCV were used successfully 1071
1010 immediately delivered to a specialized or authorized to discriminate 13 viruses from different teleost fishes 1072
1011 laboratory. cross-reacting in the SVCV/PFR-IF. In addition, RPA 1073
1012 Spring viremia of carp is listed as a notifiable disease results revealed genetic diversity among SVCV iso- 1074
1013 by the (see www.oie.int). Diagnostic procedures lates obtained from different carp species and different 1075
1014 should be based on the OIE guidelines (Office Interna- locations. Molecular methods for detection of SVCV 1076
1015 tional des Epizooties 2000). Etiological diagnosis of are not widely used at present and are not included 1077
1016 SVC is secured by the proof of virus presence in tissue among the approved methods of the OIE. However, a 1078
1017 sample using electron microscopical techniques, cryo- semi-nested PCR has been established to identify the 1079
1018 stat thin section immunofluorescence, or isolation of SVCV G gene in fish tissues (Liu et al. 1998). This 1080
1019 the agent. For the latter, cell lines such as FHM or EPC could be a method to screen for SVCV in chronic or 1081
1020 are recommended by the OIE. Cell monolayers are persistently infected fish or in fish that are carriers of 1082
1021 inoculated with 10-fold dilutions of homogenates of the virus. As RNA viruses usually form ‘quasi-species’ 1083
1022 kidney, spleen, liver and encephalon. Infected cell that include wide genomic variation, some genotypes 1084
1023 cultures incubated at 20°C usually show cytopatho- will only have a selective advantage if growth condi- 1085
1024 genic effects after 24 to 48 h. The virus yield in the tions change (Steinhauer & Holland 1987). Oreshkova 1086
1025 internal organs can vary between 104 to 107 TCID50 et al. (1995, 1999) used the reverse transcriptase 1087
1026 ml–1, depending on the phase of disease development. PCR (RT-PCR) developed for the M and G gene and 1088
1027 The isolated virus can be identified by serological hybridization with non-radioactive probes for detec- 1089
1028 techniques such as the serum neutralization test (NT), tion of SVCV. Biotinylated probes used for hybridiza- 1090
1029 immunofluorescence (IF), immunoperoxidase (IP), or tion were 248 bp fragments of the M gene. Dot blots 1091
1030 enzyme linked immunosorbent assay (ELISA) (Ahne gave sensitive virus specific signals in tissue samples 1092
1031 1981, Faisal & Ahne 1984, Jørgensen et al. 1989, of experimentally infected carp fingerlings if the 1093
1032 Way 1991, Rodak et al. 1993), although the antiserum virus titers were approximately 105 TCID50 g tissue–1. 1094
1033 used must be of high quality (Dixon & Hill 1984). In Reagents for SVCV diagnosis can be obtained from the 1095
1034 routine diagnostics, immunohistological techniques OIE reference laboratory for SVCV.1 1096
1035 using virus-infected cell cultures or a cryostat thin sec- 1097
1036 tion of infected organs are generally considered to be 1098
1037 the most reliable and convenient techniques for virus Prophylaxis and control of SVC 1099
1038 identification. Fundamental reagents are standard 1100
1039 antigen (SVCV reference strain ATCC VR-1390, Fijan Temperatures above 20°C usually secure in carp a 1101
1040 et al. 1971) and polyclonal, or preferably monoclonal, level of metabolic activity that enables production of pro- 1102
1041 antibodies (BIO-FLOU SVC, Bio-X, 30 Hoogveldiaan, tective levels of interferon (Baudoy 1978) and antibodies 1103
1042 1700 Dilbeek, Belgium). SVCV is serologically distinct (Ahne 1980). Therefore, SVC-diseased fish have so far 1104
1043 from the other known fish rhabdoviruses, with the not been reported in tropical and subtropical climates. In 1105
1044 exception of PFR that cross-reacts with SVCV by rearing facilities with a controlled environment, eleva- 1106
1045 immunofluorescence and ELISA (Jørgensen et al. tion of temperature can prevent or stop SVC outbreaks. 1107
1046 1989, Way 1991). The indirect diagnosis of SVCV is In temperate climates, avoidance and eradication seem 1108
1047 based on detection of virus-specific antibodies in the 1109
1048 fish serum. For seroepidemiological surveys, NT, IF or 1110
1049 ELISA are usually used. The immune status of investi- 1 1111
The Centre for Environment, Fisheries & Aquaculture Sci-
1050 gated fish revealed a high incidence of serologically ences, Weymouth Laboratory, Barrack Road, The Nothe, 1112
1051 positive carp farms in Europe (Ahne 1979, Dixon et al. Weymouth, Dorset DT4 8UB, UK (b.j.hill@cefas.co.uk). 1113
1052 1114
1053 1115
1054 1116
1117 10 Dis Aquat Org ■ ■ 1179
1118 1180
1119 1181
1120 1182
1121 possible on modern small farms using spring or well Ahne W (1976) Untersuchungen über die Stabilität des 1183
1122 water. On large carp farms, general prevention and con- karpfenpathogenen Virusstammes 10/3. Fisch Umwelt 2: 1184
1123 121–127 1185
trol measures have little chance of being effective.
Ahne W (1977) Evidence for the systemic character of Rhab-
1124 Location on, and connections with, large surface water dovirus carpio infection. Bull Off Int Épizoot 87:435–436 1186
1125 systems make it impossible to prevent natural move- Ahne W (1978) Uptake and multiplication of spring viremia of 1187
1126 ments of carriers such as carp and perhaps other fishes carp virus in carp, Cyprinus carpio L. J Fish Dis 1:265–268 1188
1127 from farms to open waters and back. Selection, hy- Ahne W (1979) Untersuchungen über die akute Form der 1189
infektiösen Bauchwassersucht bei Cypriniden (Cyprinus
1128 bridization and genetic manipulation of carp for SVC 1190
carpio, Ctenopharyngodon idella). Vet Med Habilschrift,
1129 resistance has not yet resulted in a tested and accepted Universität Muenchen: 1–112 1191
1130 strain. This approach to prevention has a long tradition in Ahne W (1980) Rhabdovirus carpio-Infektion beim Karpfen 1192
1131 Russia. Kirpichnikov et al. (1987) reported results of (Cyprinus carpio): Untersuchungen über Reaktionen des 1193
1132 Wirtsorganismus. Fortschr Vet 30:180–183 1194
strain selection, and Wolf (1988) mentioned the highly
Ahne W (1981) Serological techniques currently used in fish
1133 touted resistance of the Krasnodar strain, but data on virology. Dev Biol Stand 49:3–27 1195
1134 controlled challenges of such carp strains are still lack- Ahne W (1982) Vergleichende Untersuchungen über die 1196
1135 ing. Effective and safe immunoprophylaxis has not been Stabilität von vier fischpathogenen Rhabdoviren (VHSV, 1197
1136 established yet. However, carp vaccinated intraperi- PFR, SVCV, IPNV). Zentralbl Veterinärmed 29:457–476 1198
Ahne W (1985a) Argulus foliaceus L. and Piscicola geometra
1137 toneally or orally in autumn with live virus can develop a 1199
L. as mechanical vectors of spring viraemia of carp virus
1138 solid resistance to reinfection, which can last several (SVCV). J Fish Dis 8:241–242 1200
1139 months including overwintering (Fijan et al. 1977a,b, Ahne W (1985b) Viral infection cycles in pike (Esox lucius L.). 1201
1140 Fijan 1988, 1999). A commercial inactivated SVCV J Appl Ichthyol 1:90–95 1202
1141 Ahne W (1986) The influence of environmental temperature 1203
preparation for intraperitoneal delivery gave positive
and infection route on the immune response of carp
1142 results in vaccinations of carp in Eastern Europe 1204
(Cyprinus carpio) to spring viremia of carp virus. Vet
1143 (Tesar<ik & Macura 1981). A possible strategy of vacci- Immunol Immunopathol 12:383–386 1205
1144 nation and the existing limitations were reported (Fijan Ahne W, Wolf K (1977) Spring viremia of carp. US Depart- 1206
1145 & Mata$in 1980, Fijan 1984). Any type of vaccine should ment of Interior, Fish Disease Leaflet 51:1–11 1207
Ahne W, Kurath G, Winton JR (1998) A ribonuclease protec-
1146 preferably be delivered at water temperatures above 19 1208
tion assay can distinguish spring viremia of carp virus
1147 to 20°C. Broodfish can be vaccinated immediately after from pike fry rhabdovirus. Bull Eur Assoc Fish Pathol 18: 1209
1148 spawning. The 40 to 60 d old fry could be immunized by 220–224 1210
1149 bath or orally. For fingerlings in early autumn, only oral Arshaniza NM, Bauer ON, Vladimirov VL (1968) Air bladder 1211
1150 disease of carps: its aetiology, epizootiology and control. 1212
delivery is possible in the present carp production tech-
Bull Off Int Épizoot 69:999–1006
1151 nology. Depending upon local economic conditions, the 1213
Bachmann PA, Ahne W (1973) Isolation and characterization
1152 intraperitoneal vaccination of 2 yr old carp for stocking in of agent causing swim bladder inflammation of carp. 1214
1153 spring may be advantageous. More research is needed Nature 244: 235-237 1215
1154 for setting up models and standards for vaccine devel- Bachmann PA, Ahne W (1974) Biological properties and iden- 1216
tification of the agent causing swim bladder inflammation
1155 opment and for their testing in aquaculture. Although 1217
in carp. Arch Ges Virusforsch 44:261–269
1156 DNA vaccines show great promise for use against VHSV Baudouy AM (1975) Virémie printanière de la carpe: premiers 1218
1157 and IHNV (Lorenzen et al. 1999, Corbeil et al. 2000), an isolements du virus en France. Bull Off Int Épizoot 83: 1219
1158 efficacious DNA vaccine against SVCV has not yet been 717–722 1220
1159 Baudouy AM (1978) Relation hôte-virus au cours de la virémie 1221
reported, due at least in part to the difficulty of the live-
printanière de la carpe. CR Acad Sci Paris 286:1225–1228
1160 fish challenge-model in carp. However, experiments 1222
Baudouy AM, Danton M, Merle G (1980a) Virémie print-
1161 showed that plasmid DNA encoding the SVCV glyco- anière de la carpe: étude experimentale de l’infection 1223
1162 protein elicited a non-specific, protective immunity évoluant a différentes temperatures. Ann Virol (Paris) 1224
1163 against the salmonid rhabdovirus IHNV in rainbow trout 131E:479–488 1225
Baudouy AM, Danton M, Merle G (1980b) Virémie printanière
1164 (Kim et al. 2000). The nature of vaccines used in the 1226
de la carpe: résultats de contaminations experimentales
1165 future (mono- or polyvalent live or inactivated virus, effectuées au printemps. Ann Res Vét 11:245–249 1227
1166 subunit vaccine, DNA vaccine) will depend upon effec- Bauer ON, Faktorovich KA (1969) Research on infectious dis- 1228
1167 tiveness, safety, current legislation, and upon the local eases of carp in the USSR. Bul VURH Vodnany 5(3):30–43 1229
1168 Békési L, Csontos L (1985) Isolation of spring viraemia of carp 1230
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1169 1231
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1173 1235
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1242 1304
1243 1305
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1264 Corbeil S, La Patra SE, Anderson ED, Kurath G (2000) Nano- and rainbow trout. Dis Aquat Org 15:87–92 1326
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1421 Editorial responsibility: Jo-Ann Leong Submitted: November 26, 2001; Accepted: June 27, 2002 1483
Corvallis, Oregon, USA Proofs received from author(s): ❖
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