Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Diagnostic DIVA tests accompanying the Disabled Infectious Single Animal (DISA) vaccine platform for African horse sickness
Rijn, Piet van; Maris-Veldhuis, M.A. ; Boonstra, J. ; Gennip, H.G.P. van - \ 2018
Vaccine 36 (2018)25. - ISSN 0264-410X - p. 3584 - 3592.
African Horse Sickness Virus (AHSV) (Orbivirus genus, Reoviridae family) causes high mortality in naïve domestic horses with enormous economic and socio-emotional impact. There are nine AHSV serotypes showing limited cross neutralization. AHSV is transmitted by competent species of Culicoides biting midges. AHS is a serious threat beyond the African continent as endemic Culicoides species in moderate climates transmit the closely related prototype bluetongue virus. There is a desperate need for safe and efficacious vaccines, while DIVA (Differentiating Infected from Vaccinated) vaccines would accelerate control of AHS. Previously, we have shown that highly virulent AHSV with an in-frame deletion of 77 amino acids (aa) in NS3/NS3a is completely safe, does not cause viremia and shows protective capacity. This deletion mutant is a promising DISA (Disabled Infectious Single Animal) vaccine platform, since exchange of serotype specific virus proteins has been shown for all nine serotypes. Here, we show that a prototype NS3 competitive ELISA is DIVA compliant to AHS DISA vaccine platforms. Epitope mapping of NS3/NS3a shows that more research is needed to evaluate this prototype serological DIVA assay regarding sensitivity and specificity, in particular for AHSVs expressing antigenically different NS3/NS3a proteins. Further, an experimental panAHSV PCR test targeting genome segment 10 is developed that detects reference AHSV strains, whereas AHS DISA vaccine platforms were not detected. This DIVA PCR test completely guarantees genetic DIVA based on in silico and in vitro validation, although test validation regarding diagnostic sensitivity and specificity has not been performed yet. In conclusion, the prototype NS3 cELISA and the PCR test described here enable serological and genetic DIVA accompanying AHS DISA vaccine platforms.
African horse sickness virus (AHSV) with a deletion of 77 amino acids in NS3/NS3a protein is not virulent and a safe promising AHS Disabled Infectious Single Animal (DISA) vaccine platform
Rijn, Piet A. van; Maris-Veldhuis, Mieke A. ; Potgieter, Christiaan A. ; Gennip, René G.P. van - \ 2018
Vaccine 36 (2018)15. - ISSN 0264-410X - p. 1925 - 1933.
African horse sickness virus - DISA vaccine platform - NS3/NS3a protein - Pathogenic - Virulence
African horse sickness virus (AHSV) is a virus species in the genus Orbivirus of the family Reoviridae. Currently, nine serotypes have been defined showing limited cross neutralization. AHSV is transmitted by species of Culicoides biting midges and causes African Horse Sickness (AHS) in equids with a mortality up to 95% in naïve domestic horses. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates are competent vectors of closely related bluetongue virus. AHS outbreaks cause huge economic losses in developing countries. In the developed world, outbreaks will result in losses in the equestrian industry and will have an enormous emotional impact on owners of pet horses. Live-attenuated vaccine viruses (LAVs) have been developed, however, safety of these LAVs are questionable due to residual virulence, reversion to virulence, and risk on virulent variants by reassortment between LAVs or with field AHSV. Research aims vaccines with improved profiles. Reverse genetics has recently being developed for AHSV and has opened endless possibilities including development of AHS vaccine candidates, such as Disabled Infectious Single Animal (DISA) vaccine. Here, virulent AHSV5 was recovered and its high virulence was confirmed by experimental infection of ponies. 'synthetically derived’ virulent AHSV5 with an in-frame deletion of 77 amino acids codons in genome segment 10 encoding NS3/NS3a protein resulted in similar in vitro characteristics as published NS3/NS3a knockout mutants of LAV strain AHSV4LP. In contrast to its highly virulent ancestor virus, this deletion AHSV5 mutant (DISA5) was completely safe for ponies. Two vaccinations with DISA5 as well as two vaccinations with DISA vaccine based on LAV strain AHSV4LP showed protection against lethal homologous AHSV. More research is needed to further improve efficacy, to explore the AHS DISA vaccine platform for all nine serotypes, and to study the vaccine profile in more detail.
Bluetongue Disabled Infectious Single Animal (DISA) vaccine: Studies on the optimal route and dose in sheep
Rijn, P.A. van; Daus, F.J. ; Maris-Veldhuis, M.A. ; Feenstra, Femke ; Gennip, H.G.P. van - \ 2016
Vaccine 35 (2016)2. - ISSN 0264-410X - p. 231 - 237.
Bluetongue (BT) is a disease of ruminants caused by bluetongue virus (BTV) transmitted by biting midges of the Culicoides genus. Outbreaks have been controlled successfully by vaccination, however, currently available BT vaccines have several shortcomings. Recently, we have developed BT Disabled Infectious Single Animal (DISA) vaccines based on live-attenuated BTV without expression of dispensable non-structural NS3/NS3a protein. DISA vaccines are non-pathogenic replicating vaccines, do not cause viremia, enable DIVA and are highly protective. NS3/NS3a protein is involved in virus release, cytopathogenic effect and suppression of Interferon-I induction, suggesting that the vaccination route can be of importance. A standardized dose of DISA vaccine for serotype 8 has successfully been tested by subcutaneous vaccination. We show that 10 and 100 times dilutions of this previously tested dose did not reduce the VP7 humoral response. Further, the vaccination route of DISA vaccine strongly determined the induction of VP7 directed antibodies (Abs). Intravenous vaccination induced high and prolonged humoral response but is not practical in field situations. VP7 seroconversion was stronger by intramuscular vaccination than by subcutaneous vaccination. For both vaccination routes and for two different DISA vaccine backbones, IgM Abs were rapidly induced but declined after 14 days post vaccination (dpv), whereas the IgG response was slower. Interestingly, intramuscular vaccination resulted in an initial peak followed by a decline up to 21 dpv and then increased again. This second increase is a steady and continuous increase of IgG Abs. These results indicate that intramuscular vaccination is the optimal route. The protective dose of DISA vaccine has not been determined yet, but it is expected to be significantly lower than of currently used BT vaccines. Therefore, in addition to the advantages of improved safety and DIVA compatibility, the novel DISA vaccines will be cost–competitive to commercially available live attenuated and inactivated vaccines for Bluetongue.
Experimental infection of small ruminants with bluetongue virus expressing Toggenburg Orbivirus proteins
Rijn, Piet A. van; Water, Sandra G.P. van de; Maris-Veldhuis, Mieke A. ; Gennip, René G.P. van - \ 2016
Veterinary Microbiology 192 (2016). - ISSN 0378-1135 - p. 145 - 151.
Bluetongue virus - Experimental infection - Goat - Protein expression - Reverse genetics - Sheep - Toggenburg Orbivirus

Bluetongue virus (BTV) is the prototype orbivirus (Reoviridae family, genus Orbivirus) consisting of more than 24 recognized serotypes or neutralization groups. Recently, new BTV serotypes in goats have been found; serotype 25 (Toggenburg Orbivirusor TOV), serotype 26 (KUW2010/02), and serotype 27 from Corsica, France. KUW2010/02 has been isolated in mammalian cells but is not replicating in Culicoides cells. TOVhas been detected in goats but could not been cultured, although TOV has been successfully passed to naïve animals by experimental infection using viremic blood. Genome segments Seg-2[VP2], Seg-6[VP5], Seg-7[VP7], and Seg-10[NS3/NS3a] expressing the respective TOV proteins were incorporated in BTV using reverse genetics, demonstrating that these TOV proteins are functional in BTV replication. Depending on the incorporated TOV proteins, in vitro replication is, however, decreased compared to the ancestor BTV, in particular by TOV-VP5. Sheep and goats were experimentally infected with BTV expressing both outer capsid proteins VP2 and VP5 of TOV, so-named ‘TOV-serotyped BTV’. Viremia was not detected in sheep, and hardly detected in goats after infection with TOV-serotyped BTV. Seroconversion by cELISA, however, was detected, suggesting that TOV-serotyped BTV replicates in small ruminants. One goat was coincidentally pregnant, and the fetus was strong PCR-positive in blood samples and several organs, which conclusively demonstrates that TOV-serotyped BTV replicates in vivo.

Bluetongue virus without NS3/NS3a expression is not virulent and protects against virulent bluetongue virus challenge.
Feenstra, F. ; Gennip, H.G.P. van; Maris-Veldhuis, M.A. ; Verheij, E. ; Rijn, P.A. van - \ 2014
Journal of General Virology 95 (2014)Pt. 9. - ISSN 0022-1317 - p. 2019 - 2029.
vaccinated animals - rna segment - serotype 8 - sheep - antibodies - particles - europe - cattle - ns3 - differentiation
Bluetongue is a disease in ruminants caused by the bluetongue virus (BTV), and is spread by Culicoides biting midges. Bluetongue outbreaks cause huge economic losses and death in sheep in several parts of the world. The most effective measure to control BTV is vaccination. However, both commercially available vaccines and recently developed vaccine candidates have several shortcomings. Therefore, we generated and tested next-generation vaccines for bluetongue based on the backbone of a laboratory-adapted strain of BTV-1, avirulent BTV-6 or virulent BTV-8. All vaccine candidates were serotyped with VP2 of BTV-8 and did not express NS3/NS3a non-structural proteins, due to induced deletions in the NS3/NS3a ORF. Sheep were vaccinated once with one of these vaccine candidates and were challenged with virulent BTV-8 3 weeks after vaccination. The NS3/NS3a knockout mutation caused complete avirulence for all three BTV backbones, including for virulent BTV-8, indicating that safety is associated with the NS3/NS3a knockout phenotype. Viraemia of vaccine virus was not detected using sensitive PCR diagnostics. Apparently, the vaccine viruses replicated only locally, which will minimize spread by the insect vector. In particular, the vaccine based on the BTV-6 backbone protected against disease and prevented viraemia of challenge virus, showing the efficacy of this vaccine candidate. The lack of NS3/NS3a expression potentially enables the differentiation of infected from vaccinated animals, which is important for monitoring virus spread in vaccinated livestock. The disabled infectious single-animal vaccine for bluetongue presented here is very promising and will be the subject of future studies.
Immunogenicity of recombinant VP2 proteins of all nive serotypes of African horse sickness virus
Kanai, T. ; Rijn, P.A. van; Maris-Veldhuis, M.A. ; Kaname, Y. ; Athmaram, T.N. ; Roy, P. - \ 2014
Vaccine 32 (2014)39. - ISSN 0264-410X - p. 4932 - 4937.
outer capsid proteins - horsesickness-virus - bluetongue virus - protective efficacy - vaccine - baculovirus - expression - challenge - responses - sheep
African horse sickness (AHS) is an equine disease with a mortality of up to 90% for susceptible horses. The causative agent AHS virus (AHSV) is transmitted by species of Culicoides. AHSV serogroup within the genus Orbivirus of the Reoviridae family consists of nine serotypes that show no or very limited cross-neutralization. Of the seven structural proteins (VP1-VP7) of AHSV, VP2 is the serotype specific protein, and the major target for neutralizing antibodies. In this report, recombinant VP2 proteins of all nine serotypes were expressed individually by the baculovirus expression system and the immunogenicity of each was studied by immunization of guinea pigs with single VP2 as well as with cocktails of VP2 proteins. Homologous neutralizing antibodies measured by 50% plaque reduction assay showed varying degrees (from 37 to 1365) of titers for different VP2 proteins. A low cross-neutralizing antibody titer was found for genetically related AHSV serotypes. Immunization with VP2 cocktails containing equal amounts of each of the VP2 proteins also triggered neutralizing antibodies albeit to lower titers (4-117) to each of the serotypes in the cocktail. This study is a first step to develop a VP2 subunit vaccine for AHS and our results indicate that VP2 subunit vaccines are feasible individually or in a multi-serotype cocktail.
VP2-serotyped live-attenuated bluetongue virus without NS3/NS3a expression provided serotype-specific protection and enables DIVA.
Feenstra, F. ; Maris-Veldhuis, M.A. ; Daus, F.J. ; Tacken, M.G.J. ; Moormann, R.J.M. ; Gennip, H.G.P. van; Rijn, P.A. van - \ 2014
Vaccine 32 (2014)52. - ISSN 0264-410X - p. 7108 - 7114.
Bluetongue virus (BTV) causes Bluetongue in ruminants and is transmitted by Culicoides biting midges. Vaccination is the most effective measure to control vector borne diseases; however, there are 26 known BTV serotypes showing little cross protection. The BTV serotype is mainly determined by genome segment 2 encoding the VP2 protein. Currently, inactivated and live-attenuated Bluetongue vaccines are available for a limited number of serotypes, but each of these have their specific disadvantages, including the inability to differentiate infected from vaccinated animals (DIVA). BTV non-structural proteins NS3 and NS3a are not essential for virus replication in vitro, but are important for cytopathogenic effect in mammalian cells and for virus release from insect cells in vitro. Recently, we have shown that virulent BTV8 without NS3/NS3a is non-virulent and viremia in sheep is strongly reduced, whereas local in vivo replication leads to seroconversion. Live-attenuated BTV6 without NS3/NS3a expression protected sheep against BTV challenge. Altogether, NS3/NS3a knockout BTV6 is a promising vaccine candidate and has been named Disabled Infectious Single Animal (DISA) vaccine. Here, we show serotype-specific protection in sheep by DISA vaccine in which only genome segment 2 of serotype 8 was exchanged. Similarly, DISA vaccines against other serotypes could be developed, by exchange of only segment 2, and could therefore safely be combined in multi-serotype cocktail vaccines with respect to reassortment between vaccine viruses. Additionally, NS3 antibody responses are raised after natural BTV infection and NS3-based ELISAs are therefore appropriate tools for DIVA testing accompanying the DISA vaccine. To enable DIVA, we developed an experimental NS3 ELISA. Indeed, vaccinated sheep remained negative for NS3 antibodies, whereas seroconversion for NS3 antibodies was associated with viremia after heterologous BTV challenge.
Bluetongue Viruses Based on Modified-Live Vaccine Serotype 6 with Exchanged Outer Shell Proteins Confer Full Protection in Sheep against Virulent BTV8
Gennip, H.G.P. van; Water, S.G.P. van de; Maris-Veldhuis, M.A. ; Rijn, P.A. van - \ 2012
PLoS One 7 (2012)9. - ISSN 1932-6203
functional-characterization - capsid protein - replication - netherlands - expression - particles - infection - epidemic - release - genome
Since 1998, Bluetongue virus (BTV)-serotypes 1, 2, 4, 9, and 16 have invaded European countries around the Mediterranean Basin. In 2006, a huge BT outbreak started after incursion of BTV serotype 8 (BTV8) in North-Western Europe. IN 2008, BTV6 and BTV11 were reported in the Netherlands and Germany, and in Belgium, respectively. In addition, Toggenburg orbivirus (TOV) was detected in 2008 in Swiss goats, which was recognized as a new serotype of BTV (BTV25). The (re-)emergency of BTV serotypes needs a rapid response to supply effective vaccines. Reverse genetics has been developed for BTV1 and more recently also for BTV6. This latter strain, BTV6/net08, is closely related to live-attenuated vaccine for serotype 6 as determined by full genome sequencing. Here, we used this strain as backbone and exchanged segment 2 and 6, respectively Seg-2 (VP2) and Seg-6 (VP5), for those of BTV serotype 1 and 8 using reverse genetics. These so-called ‘serotyped’ vaccine viruses, as mono-serotype and multi-serotype vaccine, were compared for their protective capacity in sheep. In general, all vaccinated animals developed a neutralizing antibody response against their respective serotype. After challenge at three weeks post vaccination with cell-passaged, virulent BTV8/net07 (BTV8/net07/e1/bhkp3) the vaccinated animals showed nearly no clinical reaction. Even more, challenge virus could not be detected, and seroconversion or boostering after challenge was negligible. These data demonstrate that all sheep were protected from a challenge with BTV8/net07, since sheep of the control group showed viremia, seroconversion and clinical signs that are specific for Bluetongue. The high level of cross-protection is discussed.
Development of an animal model for bluetongue virus serotype 8
Stockhofe, N. ; Gennip, H.G.P. van; Boonstra, J. ; Backx, A. ; Wright, I.M. ; Potgieter, A. ; Wieringa, T. ; Maris-Veldhuis, M.A. ; Swanenburg, M. ; Rijn, P.A. van - \ 2010
S.n.
Transmission of bovine herpesvirus 1 within and between herds on an island with a BHV1 control programme
Hage, J.J. ; Schukken, Y.H. ; Schols, H. ; Maris-Veldhuis, M.A. ; Rijsewijk, F.A.M. ; Klaassen, C.H.L. - \ 2003
Epidemiology and Infection 130 (2003)3. - ISSN 0950-2688 - p. 541 - 552.
rhinotracheitis virus - infectious-diseases - marker vaccine - risk-factors - dairy farms - calves - immunity - type-1 - reactivation - immunization
Transmission of bovine herpesvirus 1 (BHV1) within and between herds was studied on the island of Ameland, The Netherlands. There were 50 herds with 3300 head of cattle on the island. Herds were divided into three groups: (1) only containing seronegative cattle, (2) containing seronegative cattle and vaccinated seropositive cattle, and (3) containing only vaccinated cattle. All 23 herds in groups 1 and 2 were monitored. Three major outbreaks of BHV1 infections were observed due to the introduction of infectious cattle. Another major outbreak was most likely induced by reactivation of latent BHV1 in seropositive cattle. The basic reproduction ratio within these herds was estimated at least 4. Only one of these outbreaks led to three secondary outbreaks in susceptible herds in which all cattle were seronegative. These outbreaks were most likely due to respectively, direct animal contact, human transmission, and aerogenic transmission. The basic reproduction ratio between herds in this study was estimated to be 0(.)6.
Strains of bovine herpesvirus 1 that do not express an epitope on glycoprotein E in cell culture still induce antibodies that can be detected in a gE-blocking ELISA
Oirschot, J.T. van; Kaashoek, M.J. ; Maris-Veldhuis, M.A. ; Rijsewijk, F.A.M. - \ 1999
Veterinary Microbiology 65 (1999)2. - ISSN 0378-1135 - p. 103 - 113.
Two bovine herpesvirus 1 (BHV1) field strains that do not express an epitope on glycoprotein E (gE) in cell culture were inoculated into calves to examine whether their sera became positive in a gE-blocking ELISA that detects antibodies against gE. This gE-blocking ELISA uses one monoclonal antibody that is directed against the above mentioned epitope. All calves, except one, infected with these gE-epitope negative BHV1 strains, became positive in this gE-blocking ELISA, about two weeks later than in another gE-ELISA and a gB-ELISA. However, cattle infected with BHV1 strains that do express this particular gE-epitope showed a similar type of antibody responses. These findings demonstrate that BHV1 strains that do not express a particular gE-epitope in cell culture, still can induce antibodies that are detected in a blocking ELISA that measures antibodies against that epitope.
Epitopes on glycoprotein C of bovine herpesvirus-1 (BHV-1) that allow differentiation between BHV-1.1 and BHV-1.2
Rijsewijk, F.A.M. ; Kaashoek, M.J. ; Langeveld, J.P.M. ; Meloen, R. ; Judek, J. ; Bienkowska-Szewczyk, K. ; Maris-Veldhuis, M.A. ; Oirschot, J.T. van - \ 1999
Journal of General Virology 80 (1999). - ISSN 0022-1317 - p. 1477 - 1483.
Comparison of DNA application methods to reduce BRSV shedding in cattle
Schrijver, R.S. ; Langedijk, J.P.M. ; Keil, G.M. ; Middel, W.G.J. ; Maris-Veldhuis, M. ; Oirschot, J.T. van; Rijsewijk, F.A.M. - \ 1998
Vaccine 16 (1998). - ISSN 0264-410X - p. 130 - 134.
An inactivated gE-negative marker vaccine and an experimental gD-subunit vaccine reduce the incidence of bovine herpesvirus 1 infections in the field.
Bosch, J.C. ; Jong, M.C.M. de; Franken, P. ; Frankena, K. ; Hage, J.J. ; Kaashoek, M.J. ; Maris-Veldhuis, M.A. ; Noordhuizen, J.P.T.M. ; Poel, W.H.M. van der; Verhoeff, J. ; Weerdmeester, K. ; Zimmer, G.M. ; Oirschot, J.T. van - \ 1998
Vaccine 16 (1998). - ISSN 0264-410X - p. 265 - 271.
An inactivated glycoprotein E-negative vaccine and an experimental glycoprotein D-subunit vaccine against bovine herpesvirus 1 (V1) were examined for their effectiveness in a randomized, double-bline, placebo-controlled field trial comprising 130 dairy farms. The use of these marker vaccines enabled us to monitor the incidence of infections in vaccinated populations. The aims of this trial were to evaluate whether these vaccines: (1) reduce the proportion of outbreaks in dairy herds; and (2) reduced virus transmission within dairy herds and to what extent. Vaccination with either of the two vaccines significantly reduced the proportion of herds wherein an outbreak occurred as well as the virus transmission within herds, as compared to placebo-treated herds. The estimated number of secondary cases caused by one infectious animal, expressed as the reproduction ratio R, was for both vaccines significantly >1. This indicates that when BHV1 is introduced into vaccinated herds, major outbreaks may still occur
Immunization of cattle with a BHV1 vector vaccine or a DNA vaccine both coding for the G protein of BRSV
Schrijver, R.S. ; Langedijk, J.P.M. ; Keil, G.M. ; Middel, W.G.J. ; Maris-Veldhuis, M. ; Kramps, J.A. ; Oirschot, J.T. van; Rijsewijk, F.A.M. - \ 1997
Vaccine 15 (1997). - ISSN 0264-410X - p. 1908 - 1915.
An enzyme-linked immunosorbent assay to detect antibodies against glycoprotein gE of bovine herpesvirus 1 allows differentiation between infected and vaccinated cattle
Oirschot, J.T. van; Kaashoek, M.J. ; Maris-Veldhuis, M.A. ; Weerdmeester, K. ; Rijsewijk, F.A.M. - \ 1997
Journal of Virological Methods 67 (1997)1. - ISSN 0166-0934 - p. 23 - 34.
A blocking enzyme-linked immunosorbent assay (ELISA) was developed for detecting antibodies against glycoprotein gE (gE) of bovine herpesvirus 1 (BHV1). The assay is based on the use of two monoclonal antibodies directed against different antigenic domains on gE. Sera from uninfected cattle and cattle that had been repeatedly vaccinated with gE-negative marker vaccines scored negative, whereas sera from cattle naturally or experimentally infected with BHV1 field strains scored positive in the gE-ELISA. Antibodies against gE appeared in the serum around 11 days after infection. Cattle that were first vaccinated and then challenged, thus having less virus replication, also became gE-seropositive. The sensitivity and specificity of the gE-ELISA is high, and therefore the gE-ELISA is suitable for differentiating between infected cattle and vaccinated cattle with a gE-negative vaccine.
Identification of epitopes on glycoprotein C of bovine herpesvirus 1 that allow differentiation between "IBR-like" and "IPV-like" strains
Rijsewijk, F.A.M. ; Kaashoek, M.J. ; Langeveld, J.P.M. ; Meloen, R. ; Judek, J. ; Bienkowska-Szewczyck, K. ; Maris-Veldhuis, M.A. ; Oirschot, J.T. van - \ 1997
In: Proc. int. herpesvirus workshop, 1997
Bovine herpesvirus 1 marker vaccines reduce the incidence of infections.
Bosch, J.G. ; Frankena, K. ; Franken, P. ; Hage, J.J. ; Jong, M.C.M. de; Kaashoek, M.J. ; Maris-Veldhuis, M.A. ; Noordhuizen, J.P.T.M. ; Poel, W.H.M. van der; Verhoeff, J. ; Weerdmeester, K. ; Zimmer, G.M. ; Oirsch, J.T. van - \ 1996
In: Proc. 19th World Buiatric Congress, Edinburgh UK, Vol. I - p. 42 - 44.
An inactivated vaccine based on a glycoprotein E-negative strain of bovine herpesvirus 1 induces protective immunity and allows serological differentiation
Kaashoek, M.J. ; Moerman, A. ; Madic, J. ; Rijsewijk, F.A.M. ; Weerdmeester, K. ; Maris-Veldhuis, M. ; Oirschot, J.T. van - \ 1995
Vaccine 13 (1995). - ISSN 0264-410X - p. 342 - 346.
Bovine herpes virus 1 marker vaccines reduce the incidence of infections.
Bosch, J.C. ; Frankena, K. ; Franken, P. ; Hage, J.J. ; Jong, M.C.M. de; Kaashoek, M.J. ; Maris-Veldhuis, M.A. ; Noordhuizen, J.P.T.M. ; Poel, W.H.M. van der; Verhoeff, J. - \ 1995
In: Proc. Symp. on IBR and other ruminant herpesvirus infections, Belgium - p. 50 - 50.
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