This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Costes, B.
Right arrow Articles by Vanderplasschen, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Costes, B.
Right arrow Articles by Vanderplasschen, A.

 Previous Article  |  Next Article 

Journal of Virology, April 2009, p. 2819-2830, Vol. 83, No. 7
0022-538X/09/$08.00+0     doi:10.1128/JVI.02305-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

The Major Portal of Entry of Koi Herpesvirus in Cyprinus carpio Is the Skin{triangledown}

B. Costes,1,{dagger} V. Stalin Raj,1,{dagger} B. Michel,1 G. Fournier,1 M. Thirion,1 L. Gillet,1 J. Mast,2 F. Lieffrig,3 M. Bremont,4 and A. Vanderplasschen1*

Immunology-Vaccinology (B43b), Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium,1 Department Biocontrole, Research Unit Electron Microscopy, Veterinary and Agrochemical Research Centre, VAR-CODA-CERVA, Groeselenberg 99, B-1180 Ukkel, Belgium,2 CERgroupe, rue du Carmel 1, B-6900 Marloie, Belgium,3 Unit of Molecular Virology and Immunology, INRA, CRJ Domaine de Vilvert, 78352 Jouy en Josas, France4

Received 4 November 2008/ Accepted 12 January 2009

Koi herpesvirus (KHV), recently designated Cyprinid herpesvirus 3, is the causative agent of a lethal disease in koi and common carp. In the present study, we investigated the portal of entry of KHV in carp by using bioluminescence imaging. Taking advantage of the recent cloning of the KHV genome as a bacterial artificial chromosome (BAC), we produced a recombinant plasmid encoding a firefly luciferase (LUC) expression cassette inserted in the intergenic region between open reading frame (ORF) 136 and ORF 137. Two viral strains were then reconstituted from the modified plasmid, the FL BAC 136 LUC excised strain and the FL BAC 136 LUC TK revertant strain, including a disrupted and a wild-type thymidine kinase (TK) locus, respectively. In vitro, the two recombinant strains replicated comparably to the parental FL strain. The FL BAC 136 LUC TK revertant strain was shown in vitro to induce a bioluminescent signal allowing the detection of single positive cells as early as 24 h postinfection, while in vivo, it induced KHV infection in carp that was indistinguishable from that induced by the parental FL strain. To identify the KHV portal of entry, carp were analyzed by bioluminescence imaging at different times postinfection with the FL BAC 136 LUC TK revertant strain. These analyses demonstrated that the skin of the fish covering the fins and also the body is the major portal of entry for KHV in carp. Finally, to further demonstrate the role of the skin as the KHV portal of entry, we constructed an original system, nicknamed "U-tube," to perform percutaneous infection restricted to the posterior part of the fish. All the data obtained in the present study demonstrate that the skin, and not the gills, is the major portal of entry for KHV in carp.

* Corresponding author. Mailing address: Immunology-Vaccinology (B43b), Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium. Phone: 32-4-366 42 64. Fax: 32-4-366 42 61. E-mail: A.vdplasschen{at}ulg.ac.be

{triangledown} Published ahead of print on 19 January 2009.

{dagger} B.C. and V.S.R. contributed equally to this work.

Journal of Virology, April 2009, p. 2819-2830, Vol. 83, No. 7
0022-538X/09/$08.00+0     doi:10.1128/JVI.02305-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Minamoto, T., Honjo, M. N., Kawabata, Z. (2009). Seasonal Distribution of Cyprinid Herpesvirus 3 in Lake Biwa, Japan. Appl. Environ. Microbiol. 75: 6900-6904 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»