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See detailHAEMATOLOGICAL PROFILES OF CALVES BELONGING TO HERDS WITH BOVINE NEONATAL PANCYTOPENIA HISTORY IN AND AROUND WALLONIA (BELGIUM).
Ronzoni, Anna ULg; Theron, Léonard ULg; Bayrou, Calixte ULg et al

in Buiatrissima, 8th ECBHM Symposium, 28-30 August 2013, Bern, Proceedings (2013, August)

The objective of the present study was to verify the hypothesis of subclinical BNP cases, by random sampling and haematological analysis in different herds with BNP history, in order to obtain a better ... [more ▼]

The objective of the present study was to verify the hypothesis of subclinical BNP cases, by random sampling and haematological analysis in different herds with BNP history, in order to obtain a better epidemiological picture of this disease. [less ▲]

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See detailBovine lymphotropic herpesvirus detected in Belgium
Garigliany, Mutien-Marie ULg; Bayrou, Calixte ULg; Cassart, Dominique ULg et al

in Veterinary Record : Journal of the British Veterinary Association (2013), 172

The bovine lymphotropic herpesvirus was detected for the first time outside the US, Canada and UK in a cow with nonresponsive chronic metritis.

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See detailSchmallenberg virus circulation in Belgium in 2012
Bayrou, Calixte ULg; Garigliany, Mutien-Marie ULg; Cassart, Dominique ULg et al

in Veterinary Record : Journal of the British Veterinary Association (2013), 172

Field and laboratory observations suggest that Schmallenberg virus was circulating in Belgium during the summer 2012 despite a very high herd immunity. Further studies will be conducted to determine ... [more ▼]

Field and laboratory observations suggest that Schmallenberg virus was circulating in Belgium during the summer 2012 despite a very high herd immunity. Further studies will be conducted to determine whether we are observing the last cases of this epizootic or if the above calves announce a transition to endemicity. [less ▲]

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See detailIllumination of murine gammaherpesvirus-68 cycle reveals a sexual transmission route from females to males in laboratory mice.
Francois, Sylvie; Vidick, Sarah ULg; Sarlet, Mickael et al

in PLoS Pathogens (2013), 9(4), 1003292

Transmission is a matter of life or death for pathogen lineages and can therefore be considered as the main motor of their evolution. Gammaherpesviruses are archetypal pathogenic persistent viruses which ... [more ▼]

Transmission is a matter of life or death for pathogen lineages and can therefore be considered as the main motor of their evolution. Gammaherpesviruses are archetypal pathogenic persistent viruses which have evolved to be transmitted in presence of specific immune response. Identifying their mode of transmission and their mechanisms of immune evasion is therefore essential to develop prophylactic and therapeutic strategies against these infections. As the known human gammaherpesviruses, Epstein-Barr virus and Kaposi's Sarcoma-associated Herpesvirus are host-specific and lack a convenient in vivo infection model; related animal gammaherpesviruses, such as murine gammaherpesvirus-68 (MHV-68), are commonly used as general models of gammaherpesvirus infections in vivo. To date, it has however never been possible to monitor viral excretion or virus transmission of MHV-68 in laboratory mice population. In this study, we have used MHV-68 associated with global luciferase imaging to investigate potential excretion sites of this virus in laboratory mice. This allowed us to identify a genital excretion site of MHV-68 following intranasal infection and latency establishment in female mice. This excretion occurred at the external border of the vagina and was dependent on the presence of estrogens. However, MHV-68 vaginal excretion was not associated with vertical transmission to the litter or with horizontal transmission to female mice. In contrast, we observed efficient virus transmission to naive males after sexual contact. In vivo imaging allowed us to show that MHV-68 firstly replicated in penis epithelium and corpus cavernosum before spreading to draining lymph nodes and spleen. All together, those results revealed the first experimental transmission model for MHV-68 in laboratory mice. In the future, this model could help us to better understand the biology of gammaherpesviruses and could also allow the development of strategies that could prevent the spread of these viruses in natural populations. [less ▲]

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See detailFunctional characterization of new allelic polymorphisms identified in the promoter region of the human MxA gene.
Tran Thi Duc, Tam; Desmecht, Daniel ULg; Cornet, Anne

in International Journal of Immunogenetics (2013), 40

The Mx proteins are high-molecular-weight dynamin-like proteins whose expression depends strictly on type-I and -III interferons (IFN). Some isoforms are able to inhibit the life cycle of one or several ... [more ▼]

The Mx proteins are high-molecular-weight dynamin-like proteins whose expression depends strictly on type-I and -III interferons (IFN). Some isoforms are able to inhibit the life cycle of one or several viruses and are thus components of innate immune response. The human MxA protein displays the broadest antiviral spectrum which makes it appear as a key antiviral effector of innate immunity. Allelic polymorphisms located in the MxA gene promoter can be expected to affect the magnitude of MxA mRNA transcription in response to IFNs and therefore to alter the severity of viral diseases in humans. Here, three single nucleotide polymorphism sites (-309, -101 and +20) were examined for their ability to alter MxA gene promoter-driven reporter expression. We show that, besides the previously reported role of 123A and -88T, the presence of -101G is equally important. Moreover, when a promoter construct carries these three critical nucleotides, a first additional positive effect is conferred by a C at position -309 and, in this latter case, a second additional effect is produced by a A at position +20. This finding is clinically useful to improve prediction of IFN-responsiveness in patients not only with viral diseases for which type-I IFN therapy is used. [less ▲]

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See detailBluetongue Virus RNA Detection by Real-Time RT-PCR in Post-Vaccination Samples from Cattle.
De Leeuw, I.; Garigliany, Mutien-Marie ULg; Bertels, G. et al

in Transboundary and Emerging Diseases (2013)

Bluetongue virus serotype 8 (BTV-8) was responsible for a large outbreak among European ruminant populations in 2006-2009. In spring 2008, a massive vaccination campaign was undertaken, leading to the ... [more ▼]

Bluetongue virus serotype 8 (BTV-8) was responsible for a large outbreak among European ruminant populations in 2006-2009. In spring 2008, a massive vaccination campaign was undertaken, leading to the progressive disappearance of the virus. During surveillance programmes in Western Europe in 2010-2011, a low but significant number of animals were found weakly positive using BTV-specific real-time RT-PCR, raising questions about a possible low level of virus circulation. An interference of the BTV-8 inactivated vaccine on the result of the real-time RT-PCR was also hypothesized. Several studies specifically addressed the potential association between a recent vaccination and BTV-8 RNA detection in the blood of sheep. Results were contradictory and cattles were not investigated. To enlighten this point, a large study was performed to determine the risks of detection of bluetongue vaccine-associated RNA in the blood and spleen of cattle using real-time RT-PCR. Overall, the results presented clearly demonstrate that vaccine viral RNA can reach the blood circulation in sufficient amounts to be detected by real-time RT-PCR in cattle. This BTV-8 vaccine RNA carriage appears as short lasting. [less ▲]

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See detailEpizootic spread of emerging Schmallenberg virus in wild cervids, Belgium, fall 2011
Linden, Annick ULg; Desmecht, Daniel ULg; Volpe, Rosario ULg et al

in Emerging Infectious Diseases (2012), 18(12), 2006-2008

The Schmallenberg virus emerged in summer-fall 2011 in North-West Europe. During the fall of 2011, the virus widely spread in red and roe deer populations living about 250 km from the emergence location.

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See detailProtective role of P2Y(2) receptor against lung infection induced by pneumonia virus of mice
Vanderstocken, Gilles; Van de Paar, Els; Robaye, Benoit et al

in PLoS ONE (2012), 7(11), 50385

ATP released in the early inflammatory processes acts as a danger signal by binding to purinergic receptors expressed on immune cells. A major contribution of the P2Y2 receptor of ATP/UTP to dendritic ... [more ▼]

ATP released in the early inflammatory processes acts as a danger signal by binding to purinergic receptors expressed on immune cells. A major contribution of the P2Y2 receptor of ATP/UTP to dendritic cell function and Th2 lymphocyte recruitment during asthmatic airway inflammation was previously reported. We investigated here the involvement of P2Y2 receptor in lung inflammation initiated by pneumonia virus of mice infection. We demonstrated that P2Y2-/- mice display a severe increase in morbidity and mortality rate in response to the virus. Lower survival of P2Y2-/- mice was not correlated with excessive inflammation despite the higher level of neutrophil recruiters in their bronchoalveolar fluids. Interestingly, we observed reduced ATP level and lower numbers of dendritic cells, CD4+ T cells and CD8+ T cells in P2Y2-/- compared to P2Y2+/+ infected lungs. Lower level of IL-12 and higher level of IL-6 in bronchoalveolar fluid support an inhibition of Th1 response in P2Y2-/- infected mice. Quantification of DC recruiter expression revealed comparable IP-10 and MIP-3 levels but a reduced BRAK level in P2Y2-/- compared to P2Y2+/+ bronchoalveolar fluids. Higher morbidity and mortality of P2Y2-/- mice appear to result from defective dendritic cell and T cell infiltration that were correlated with higher virus titer. In conclusion, P2Y2 receptor previously described as a target in cystic fibrosis therapy and as a mediator of Th2 response in asthma, may also regulate Th1 response protecting mice against lung viral infection. [less ▲]

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See detailCharacterization of the resistance of SJL/J mice to pneumonia virus of mice, a model for infantile bronchiolitis due to a respiratory syncytial virus
Glineur, Stéphanie ULg; bui tran anh, dao; Sarlet, Michaël ULg et al

in PLoS ONE (2012), 7(10), 44581

Respiratory syncytial virus (RSV), a prominent cause of airway morbidity in children, maintains an excessive hospitalization rate despite decades of research. Host factors are assumed to influence the ... [more ▼]

Respiratory syncytial virus (RSV), a prominent cause of airway morbidity in children, maintains an excessive hospitalization rate despite decades of research. Host factors are assumed to influence the disease severity. As a first step toward identifying the underlying resistance mechanisms, we recently showed that inbred mouse strains differ dramatically as regards their susceptibility to pneumonia virus of mice (PVM), the murine counterpart of RSV. PVM infection in mice has been shown to faithfully mimic the severe RSV disease in human infants. This study aimed at dissecting the remarkable PVM-resistance shown by the SJL/J strain. To characterize its genetic component, we assessed clinical, physiopathological, and virological resistance/susceptibility traits in large first (F1) and second (F2) generations obtained by crossing the SJL/J (resistant) and 129/Sv (susceptible) strains. Then, to acquire conclusive in vivo evidence in support of the hypothesis that certain radiosensitive hematopoietic cells might play a significant role in PVM-resistance, we monitored the same resistance/susceptibility traits in mock- and γ-irradiated SJL/J mice. Segregation analysis showed that (i) PVM-resistance is polygenic, (ii) the resistance alleles are recessive, and (iii) all resistance-encoding alleles are concentrated in SJL/J. Furthermore, there was no alteration of SJL/J PVM resistance after immunosuppression by γ-irradiation, which suggests that adaptive immunity is not involved. We conclude that host resistance to pneumoviruses should be amenable to genetic dissection in this mouse model and that radioresistant lung epithelial cells and/or alveolar macrophages may control the clinical severity of pneumovirus-associated lung disease. [less ▲]

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See detailPreliminary results on in vitro murine norovirus susceptibility to Mx-mediated innate immunity
Mauroy, Axel ULg; Garigliany, Mutien-Marie ULg; Desmecht, Daniel ULg et al

Poster (2012, September)

Objectives Murine norovirus (MuNoV) belongs to the family Caliciviridae, genus Norovirus and is currently used as study model for human noroviruses, major aetiologic agents of gastroenteritis worldwide ... [more ▼]

Objectives Murine norovirus (MuNoV) belongs to the family Caliciviridae, genus Norovirus and is currently used as study model for human noroviruses, major aetiologic agents of gastroenteritis worldwide. Myxovirus (Mx) protein is an interferon-induced protein that host cell can oppose virus infection and was detected in several species including human being. In mice, two genes encoding Mx1 and 2 proteins are present but it was evidenced that these genes were inactivated by deletions in several laboratory mouse strains. Mx antiviral activity was detected on several negative stranded RNA viruses but information are still lacking for most of positive stranded RNA viruses. In this study, the susceptibility of the MuNoV to specific and interspecific Mx proteins was investigated. Methods RAW264.7 cells (murine macrophages) were first tested for constitutive expression of Mx1 proteins. Plasmids containing the murine Mx1, bovine Mx1 and human MxA genes under the CMV immediate early promotor were then used to transfect RAW264.7 cells for transient Mx expression. Negative control consisted in a plasmid expressing eGFP. Four hours after transfection, cells were infected at low MOI with the CW1 MuNoV strain. Cells and supernatants were harvested 24h post infection. RNA was extracted and viral genomic copies were measured by real time RT-PCR. Results An effect was confirmed on CW1 replication for both specific and interspecific Mx proteins. The highest effect was obtained with the bovine Mx1 protein. Conclusion In conclusion, we showed in these preliminary in vitro studies the MuNoV susceptibility to specific and interspecific Mx proteins. Bovine Mx1 protein was already demonstrated to have important antiviral activity on negative stranded RNA viruses (influenza- and paramyxoviruses) and co-evolution with the host could explain a higher susceptibility to interspecific Mx proteins. Important implications of this adaptation could be expected on zoonotic concerns associated to NoV. Moreover, even if several control studies are still be conducted to validate these preliminary results, they could drive several pertaining questions on the MuNoV model used with laboratory mice. Perspectives of this work consist to validate the susceptibility in vivo and also to test the murine Mx2 antiviral activity on MuNoV. [less ▲]

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See detailSchmallenberg virus in domestic cattle, Belgium, 2012
Garigliany, Mutien-Marie ULg; Bayrou, Calixte ULg; Kleijnen, Déborah ULg et al

in Emerging Infectious Diseases (2012), 18(9), 1512-1514

The Schmallenberg virus emerged in summer-fall 2011 in North-West Europe. Nine months later, 91% of adult cattle living about 250 km from the emergence location tested positive for IgGs targeting the new ... [more ▼]

The Schmallenberg virus emerged in summer-fall 2011 in North-West Europe. Nine months later, 91% of adult cattle living about 250 km from the emergence location tested positive for IgGs targeting the new virus nucleoprotein. Further, the risk of infection of the fetus in an immunologically naive herd is 28%. [less ▲]

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See detailDetection of new biallelic polymorphisms in the human MxA gene
Tran Thi Duc, Tam; Farnir, Frédéric ULg; Michaux, Charles ULg et al

in Molecular Biology Reports (2012), 39(8), 8533-8538

The interferon-inducible human MxA protein plays an important role in innate defense against an array of viruses. One might expect allelic diversity at the MxA locus to influence the timing and magnitude ... [more ▼]

The interferon-inducible human MxA protein plays an important role in innate defense against an array of viruses. One might expect allelic diversity at the MxA locus to influence the timing and magnitude of its expression or even the range of viruses whose biological cycle is inhibited by the encoded product. Here we have collected 267 samples of genomic DNA from three distinct populations (European, Asian, and African) and have systematically sequenced the promoter of the MxA gene and its 17 exons in order to inventory its allelic variants. Fifteen single-nucleotide polymorphisms were detected, four of which had never been identified before. Two of these, located in the promoter (at positions -309 and -101 respectively), might affect the MxA expression pattern. The other two result in substitutions (Gly255Glu and Val268Met) in the protein’s N-terminal region that might directly affect its antiviral function. [less ▲]

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See detailIn vivo modulation of the innate response to pneumovirus by type-I and -III interferon-induced Bos taurus Mx1
Dermine, Martin ULg; Desmecht, Daniel ULg

in Journal of Interferon & Cytokine Research (2012), 32(7), 332-337

The respiratory syncytial virus (RSV) is a major pathogen of the human species. This pneumovirus is a prominent cause of airway morbidity in children and maintains an excessive hospitalization rate ... [more ▼]

The respiratory syncytial virus (RSV) is a major pathogen of the human species. This pneumovirus is a prominent cause of airway morbidity in children and maintains an excessive hospitalization rate despite decades of research. As involvement of a genetic vulnerability is a possibility supported by recent data, we addressed the question of whether the Mx gene products, the typical target of which consists in single-stranded negative-polarity RNA viruses, could alter the course of pneumovirus-associated disease in vivo. Wild-type and Bos taurus Mx1-expressing transgenic FVB/J mice were inoculated with the mouse counterpart and closest phylogenetic relative of RSV, pneumonia virus of mice. Survival data and follow-up of body weight, histological scores, lung virus spread and lung viral load unequivocally showed that the viral infection was severely repressed in Mx-transgenic mice, thus suggesting that pneumoviruses belong to the antiviral spectrum of mammalian Mx GTPases. Elucidating the underlying mechanisms at the molecular level could reveal critical information for the development of new anti-RSV molecules. [less ▲]

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See detailHuman/bovine chimeric MxA-like GTPases reveal a contribution of N-terminal domains to the magnitude of anti-influenza A activity
Garigliany, Mutien-Marie ULg; Cornet, Anne ULg; Desmecht, Daniel ULg

in Journal of Interferon & Cytokine Research (2012), 32(7), 326-331

Type I interferons (IFN-) provide powerful and universal innate intracellular defence mechanisms against viruses. Among the antiviral effectors induced by IFN-, Mx proteins of some species appear as ... [more ▼]

Type I interferons (IFN-) provide powerful and universal innate intracellular defence mechanisms against viruses. Among the antiviral effectors induced by IFN-, Mx proteins of some species appear as key components of defence against influenza A viruses. The body of work published to date suggests that to exert anti-influenza activity, an Mx protein must possess a GTP-binding site, structural bases allowing multimerisation, and a specific C-terminal GTPase effector domain (GED). The human MxA and bovine Mx1 proteins both meet these minimal requirements, but the bovine protein is more active against influenza viruses. Here we measured the anti-influenza activity exerted by two human/bovine chimeric Mx proteins. We show that substituting the bovine GED for the human one in human MxA does not affect the magnitude of anti-influenza activity. Strikingly, however, substituting the human GED for the bovine one in bovine Mx1 yields a chimeric protein with much higher anti-influenza activity than the human protein. We conclude, in contradiction to the hypothesis currently in vogue in the literature, that the GED is not the sole determinant controlling the magnitude of the anti-influenza activity exercised by an Mx protein that can bind GTP and multimerise. Our results suggest that one or several motifs that remain to be discovered, located N-terminally with respect to the GED, may interact with a viral component or a cellular factor so as to alter the viral cycle. Identifying, in the N-terminal portion of bovine Mx1, the motif(s) responsible for its higher anti-influenza activity could contribute to the development of new anti-influenza molecules. [less ▲]

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See detailSchmallenberg virus: a new Shamonda/Sathuperi-like virus on the rise in Europe
Garigliany, Mutien-Marie ULg; Bayrou, Calixte ULg; Kleijnen, Déborah ULg et al

in Antiviral Research (2012), 95

In the summer-fall of 2011, a nonspecific febrile syndrome characterized by hyperthermia, drop in milk production and watery diarrhea was reported in adult dairy cows from a series of farms located in ... [more ▼]

In the summer-fall of 2011, a nonspecific febrile syndrome characterized by hyperthermia, drop in milk production and watery diarrhea was reported in adult dairy cows from a series of farms located in North-West Europe. Further, in November 2011, an enzootic outbreak of abortion, stillbirth and birth at term of lambs, kids and calves with neurologic signs and/or head, spine or limb malformations emerged throughout several European countries. Both syndromes were associated with the presence in the blood (adults) or in the central nervous system (newborns) of the genome of a new Shamonda-like orthobunyavirus provisionally named Schmallenberg virus after the place where the first positive samples were collected. The clinical, pathological, virological and epidemiological facts that were made publicly available during the first 6 months after the emergence are presented here. Current knowledge of the epidemiology of the phylogenetically closest relatives of the newcomer (Shamonda, Aino and Akabane viruses) is not exhaustive enough to predict whether the current outbreak of Schmallenberg virus is the prelude to endemicity or to a 2 years long outbreak before the infection burns out when serologically naïve animals are no longer available. In the future, cyclic epizootic reemergences are a possibility too, either synchronized with a global decrease of herd immunity or due to antigenic variants escaping the immunity acquired against their predecessors. The latter hypothesis seems unlikely because of the wide array of biologic constraints acting on the genome of viruses whose life cycle requires transmission by a vector, which represses genetic drift. The remarkable stability of the Shamonda virus genome over the last forty years is reassuring in this regard. [less ▲]

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See detailGranulomatous meningo-encephalitis caused by Toxoplasma gondii in three bulls, a possible explanation for unexplained sporadic bovine meningo-encephalitis
Theron, Léonard ULg; Tabaran, F; Cassart, Dominique ULg et al

in Revista Portuguesa de Buiatria (2012, June), (Special Edition),

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See detailDetection of disease resistance and susceptibility alleles in pigs using oligonucleotide microarray hybridization
Pastoret, Soumya; Ameels, Hélène; Bossiroy, Frédérique et al

in Journal of Veterinary Diagnostic Investigation (2012), 24(3), 479-488

A multiplex DNA microarray chip aimed at the identification of allelic polymorphisms was developed for simultaneous detection of swine disease resistance genes underlying malignant hyperthermia (RYR ... [more ▼]

A multiplex DNA microarray chip aimed at the identification of allelic polymorphisms was developed for simultaneous detection of swine disease resistance genes underlying malignant hyperthermia (RYR ), postweaning diarrhea, edema disease (FUT1), neonatal diarrhea (MUC4), and influenza (MX1). The on-chip detection was performed with fragmented polymerase chain reaction (PCR)-amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required. The targets were biotin labeled during the PCR reaction, and the arrays were detected using a colorimetric methodology. Target recognition was provided by specific capture probes designed for each susceptible or resistant allelic variant. Sequencing was chosen as the golden standard to assess chip’s accuracy. All genotypes retrieved from the microarray (476) fitted with sequencing data in spite of the fact that each pig was heterozygote for at least 1 target gene. [less ▲]

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See detailSchmallenberg virus in calf born at term with porencephaly, Belgium
Garigliany, Mutien-Marie ULg; Hoffmann, Bernd; Dive, Marc et al

in Emerging Infectious Diseases (2012), 18(6), 1005-1006

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See detailGenital re-excretion of Murid gammaherpesvirus 4 following intranasal infection
François, Sylvie ULg; Vidick, Sarah ULg; Sarlet, Michaël ULg et al

in Proceedings of the 1st Scientific Meeting of the Faculty of Veterinary Medicine (2011, December 09)

Gammaherpesviruses are the archetypes of persistent viruses that have been identified in a range of animals from mice to man. As the human gammaviruses have no well-established in vivo infection model ... [more ▼]

Gammaherpesviruses are the archetypes of persistent viruses that have been identified in a range of animals from mice to man. As the human gammaviruses have no well-established in vivo infection model, related animal gammaherpesviruses are an important source of information. We are studying Murid herpesvirus 4 (MuHV-4) in inbred laboratory mouse strains which are commonly accepted as a good model for studying gammaherpesviruses in vivo. To date, it has however never been possible to monitor viral reexcretion and virus transmission in this species. In order to identify potential re-excretion sites, intranasally infected mice were followed through global luciferase imaging for up to six months after infection. Surprisingly, we detected transient viral replication in mice genital tract at various times after latency establishment. Ex vivo imaging, quantitative PCR and immunohistochemistry revealed that virus genomes were present in high quantity in the vaginal tissue and that viral replication occurred mainly at the vaginal external border. Moreover, we highlighted the presence of free infectious viruses in the vaginal cavity at the moment of the observation of viral replication. As this ephemeral viral reexcretion could reveal a link with reproductive cycle, we compared reexcretion in normal and ovariectomized mice. Interestingly, no viral reactivation was observed in absence of hormonal cycle. In conclusion, we experimentally indentified for the first time a reexcretion site for MuHV-4 in mice that had been intranasaly infected. In the future, these results could help us to better understand the biology of gammaherpesviruses but should also allow us to develop strategies that could prevent the spread of these viruses in natural populations. [less ▲]

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