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Comparative analysis of replication characteristics of different BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment

Lennert Steukers (UGent) , Sarah Glorieux (UGent) , Annelies Vandekerckhove (UGent) and Hans Nauwynck (UGent)
Author
Organization
Abstract
Bovine herpes virus 1 (BoHV-1) belongs to the subfamily of the Alphaherpesvirinae. In general, alphaherpesviruses use the epithelium of the upper respiratory and/or genital tract as a preferential site for primary replication. BoHV-1 may replicate at both sites and causes two major clinical entities designated infectious bovine rhinotracheïtis (IBR) and infectious pustular vulvovaginitis/balanoposthitis (IPV/IPB) in cattle. It has been hypothesized that subtype 1.1 invades preferentially the upper respiratory mucosa where subtype 1.2 favors replication at the peripheral genital tract. However, some in vivo data are in contrast with that hypothesis. A thorough study of primary replication at both mucosae could elucidate this question. In the present study, we established a bovine respiratory and genital organ culture. The effect of in vitro culture on tissue viability and tissue morphometry was extensively assessed by means of TUNEL stainings, light microscopy, scanning electron microscopy and transmission electron microscopy. No significant changes in viability and morphometry were observed during in vitro culture. Next, bovine respiratory and genital mucosal explants of the same animals were inoculated with several BoHV-1 subtypes. A quantitative analysis of viral mucosal invasion was performed at 0, 24, 48 and 72 hours post inoculation (pi) by measuring plaque latitude and penetration depth underneath the basement membrane (BM). BoHV-1 spread plaquewise in the epithelium and penetrated the BM. Viral plaques were observed at 24hpi for all isolates in both respiratory and genital mucosa explants with the plaque latitude increasing over time. At 24hpi, all the different subtypes spread more efficiently lateral in the epithelium of the genital tract compared to the respiratory tract. On the contrary, all BoHV-1 subtypes exhibited a more profound invasion capacity in the respiratory tract at 24h pi compared to the genital tract. These findings take the edge of the belief of the existence of specific mucosa tropisms of different BoHV-1 subtypes. This research was supported by the Agency for Innovation by Science and Technology in Flanders (IWT).

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MLA
Steukers, Lennert, Sarah Glorieux, Annelies Vandekerckhove, et al. “Comparative Analysis of Replication Characteristics of Different BoHV-1 Subtypes in Bovine Respiratory and Genital Mucosa Explants: a Phylogenetic Enlightenment.” Belgian Society for Microbiology, Meeting, Abstracts. 2010. 80–80. Print.
APA
Steukers, L., Glorieux, S., Vandekerckhove, A., & Nauwynck, H. (2010). Comparative analysis of replication characteristics of different BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment. Belgian Society for Microbiology, Meeting, Abstracts (pp. 80–80). Presented at the Meeting of the Belgian Society for Microbiology : Molecular dialogue in host-parasite interaction.
Chicago author-date
Steukers, Lennert, Sarah Glorieux, Annelies Vandekerckhove, and Hans Nauwynck. 2010. “Comparative Analysis of Replication Characteristics of Different BoHV-1 Subtypes in Bovine Respiratory and Genital Mucosa Explants: a Phylogenetic Enlightenment.” In Belgian Society for Microbiology, Meeting, Abstracts, 80–80.
Chicago author-date (all authors)
Steukers, Lennert, Sarah Glorieux, Annelies Vandekerckhove, and Hans Nauwynck. 2010. “Comparative Analysis of Replication Characteristics of Different BoHV-1 Subtypes in Bovine Respiratory and Genital Mucosa Explants: a Phylogenetic Enlightenment.” In Belgian Society for Microbiology, Meeting, Abstracts, 80–80.
Vancouver
1.
Steukers L, Glorieux S, Vandekerckhove A, Nauwynck H. Comparative analysis of replication characteristics of different BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment. Belgian Society for Microbiology, Meeting, Abstracts. 2010. p. 80–80.
IEEE
[1]
L. Steukers, S. Glorieux, A. Vandekerckhove, and H. Nauwynck, “Comparative analysis of replication characteristics of different BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment,” in Belgian Society for Microbiology, Meeting, Abstracts, Brussels, Belgium, 2010, pp. 80–80.
@inproceedings{1153254,
  abstract     = {Bovine herpes virus 1 (BoHV-1) belongs to the subfamily of the Alphaherpesvirinae. In general, alphaherpesviruses use the epithelium of the upper respiratory and/or genital tract as a preferential site for primary replication. BoHV-1 may replicate at both sites and causes two major clinical entities designated infectious bovine rhinotracheïtis (IBR) and infectious pustular vulvovaginitis/balanoposthitis (IPV/IPB) in cattle. It has been hypothesized that subtype 1.1 invades preferentially the upper respiratory mucosa where subtype 1.2 favors replication at the peripheral genital tract. However, some in vivo data are in contrast with that hypothesis. A thorough study of primary replication at both mucosae could elucidate this question. 
In the present study, we established a bovine respiratory and genital organ culture. The effect of in vitro culture on tissue viability and tissue morphometry was extensively assessed by means of TUNEL stainings, light microscopy, scanning electron microscopy and transmission electron microscopy. No significant changes in viability and morphometry were observed during in vitro culture. Next, bovine respiratory and genital mucosal explants of the same animals were inoculated with several BoHV-1 subtypes. A quantitative analysis of viral mucosal invasion was performed at 0, 24, 48 and 72 hours post inoculation (pi) by measuring plaque latitude and penetration depth underneath the basement membrane (BM). BoHV-1 spread plaquewise in the epithelium and penetrated the BM. Viral plaques were observed at 24hpi for all isolates in both respiratory and genital mucosa explants with the plaque latitude increasing over time. At 24hpi, all the different subtypes spread more efficiently lateral in the epithelium of the genital tract compared to the respiratory tract. On the contrary, all BoHV-1 subtypes exhibited a more profound invasion capacity in the respiratory tract at 24h pi compared to the genital tract. These findings take the edge of the belief of the existence of specific mucosa tropisms of different BoHV-1 subtypes.
This research was supported by the Agency for Innovation by Science and Technology in Flanders (IWT).},
  author       = {Steukers, Lennert and Glorieux, Sarah and Vandekerckhove, Annelies and Nauwynck, Hans},
  booktitle    = {Belgian Society for Microbiology, Meeting, Abstracts},
  language     = {eng},
  location     = {Brussels, Belgium},
  pages        = {80--80},
  title        = {Comparative analysis of replication characteristics of different BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment},
  year         = {2010},
}