J.T. van Oirschot

A conventionally attenuated glycoprotein E-negative strain of bovine herpesvirus type 1 is an efficacious and safe vaccine.

We examined the vaccine properties of a conventionally attenuated bovine herpesvirus type 1 (BHV-1) mutant strain A. This strain has a deletion that encompasses the glycoprotein E gene and may therefore be used as a marker vaccine. We compared strain A with a commercial live BHV-1 vaccine B. Calves were intranasally vaccinated with strain A or with vaccine B. Strain A was virtually avirulent for young calves. After challenge, vaccinated calves were protected against disease and virus shedding was considerably reduced. After dexamethasone treatment, strain A was not recovered, whereas vaccine B and challenge virus were. We conclude that strain A is suitable for inclusion in a vaccine and has the potential advantage of being used as a marker vaccine.

Diva vaccines that reduce virus transmission.

Abstract This brief review deals with the effect of diva ( D ifferentiating I nfected from VA ccinated individuals) vaccines (also termed marker vaccines) on transmission of herpesviruses and pestiviruses in swine and cattle. Pseudorabies and bovine herpesvirus 1 diva vaccines have been demonstrated to reduce transmission of wild-type virus in populations of pigs and cattle in the laboratory as well as in the field. A subunit diva vaccine based on the immunodominant E2 protein of classical swine fever virus that is expressed in the baculovirus system may reduce transmission of wild-type virus among pigs and also transmission from mother to foetuses. A similar diva vaccine against bovine virus diarrhoea infections protected sheep against transplacental transmission of antigenically homologous wild-type virus. Diva vaccines along with their companion diagnostic tests can play a role in control of infections, ultimately leading to eradication of viruses.

Viral infections and bovine mastitis: a review

This review deals with the role of viruses in the aetiology of bovine mastitis. Bovine herpesvirus 1, bovine herpesvirus 4, foot-and-mouth disease virus, and parainfluenza 3 virus have been isolated from milk from cows with clinical mastitis. Intramammary inoculations of bovine herpesvirus 1 or parainfluenza 3 virus-induced clinical mastitis, while an intramammary inoculation of foot-and-mouth disease virus resulted in necrosis of the mammary gland. Subclinical mastitis has been induced after a simultaneous intramammary and intranasal inoculation of lactating cows with bovine herpesvirus 4. Bovine leukaemia virus has been detected in mammary tissue of cows with subclinical mastitis, but whether this virus was able to induce bovine mastitis has not been reported. Bovine herpesvirus 2, vaccinia, cowpox, pseudocowpox, vesicular stomatitis, foot-and-mouth disease viruses, and bovine papillomaviruses can play an indirect role in the aetiology of bovine mastitis. These viruses can induce teat lesions, for instance in the ductus papillaris, which result in a reduction of the natural defence mechanisms of the udder and indirectly in bovine mastitis due to bacterial pathogens. Bovine herpesvirus 1, bovine viral diarrhoea virus, bovine immunodeficiency virus, and bovine leukaemia virus infections may play an indirect role in bovine mastitis, due to their immunosuppressive properties. But, more research is warranted to underline their indirect role in bovine mastitis. We conclude that viral infections can play a direct or indirect role in the aetiology of bovine mastitis; therefore, their importance in the aetiology of bovine mastitis and their economical impact needs further attention.

Vaccination of cattle against bovine viral diarrhoea.

This brief review describes types and quality (efficacy and safety) of bovine viral diarrhoea virus (BVDV) vaccines that are in the market or under development. Both conventional live and killed vaccines are available. The primary aim of vaccination is to prevent congenital infection, but the few vaccines tested are not highly efficacious in this respect, as shown in vaccination-challenge experiments. Vaccination to prevent severe postnatal infections may be indicated when virulent BVDV strains are prevalent. Live BVDV vaccines have given rise to safety problems. A complication for the development of BVDV vaccines is the wide antigenic diversity among wild-type BVDV. There is ample room for improvement of both the efficacy and safety of BVDV vaccines, and it may be expected that better vaccines, among which marker vaccines, will be launched in the future.

Marker vaccines, virus protein-specific antibody assays and the control of Aujeszky's disease

Vaccination of pigs is widely practised to control Aujeszkys disease (AD). Molecular biological research revealed that several conventionally attenuated virus vaccines harbour deletions in their genomes. The deleted genes are nonessential for virus replication and can be involved in the expression of virulence. These findings have prompted several groups to construct well-characterized deletion mutants of AD virus that do not express either glycoprotein gI, gX or gIII. These mutants have also been rendered thymidine kinase negative. Although data on vaccine efficacy and safety have been published, widely varying test conditions have made it impossible to identify the most efficacious deletion mutant vaccine(s). Vaccination enhances the amount of virus required for infection and reduces, but does not prevent, the shedding of virulent virus and the establishment of latency in pigs infected with virulent AD virus. Therefore, while a vaccination programme will reduce the circulation of virus in the field, it will not eliminate AD virus from pig populations. To eradicate AD, the ability to differentiate infected from vaccinated pigs is crucial. The use of marker vaccines enables us to identify infected pigs in vaccinated populations by detecting antibodies against the protein whose gene is deleted from vaccine strains. The antibody response to gI appears to persist for more than 2 years, and all of about 300 field strains tested so far express gI. The use of vaccines lacking gI in combination with an enzyme linked immunosorbent assay to detect antibodies to gI and culling of gI-seropositive pigs, may help to eradicate AD in countries where vaccination is widely practised.

Respiratory syncytial virus infections in human beings and in cattle

Respiratory syncytial virus (RSV) causes yearly outbreaks of respiratory disease in human beings and cattle all over the world. Most severe human respiratory syncytial virus (HRSV)-associated disease is observed in children less than 1 year of age while most severe bovine respiratory syncytial virus (BRSV)-associated disease is observed in calves less than 6 months of age. Two subgroups of HRSV have been identified. The existence of two subgroups of BRSV has been repeatedly suggested but is not yet well established. BRSV and HRSV are closely related antigenically but antigenic differences have been observed. Seasonal periodicity of RSV infection is usual with highest incidences in autumn and winter. Stress such as caused by movement, crowding and temperature changes are considered to play a role in bovine outbreaks. Human beings and cattle are the natural hosts of HRSV and BRSV, respectively. Primarily infected individuals are the most important source of RSV during outbreaks. The role of other species in the spread of HRSV and BRSV is unknown. Protective efficacy of maternally derived antibodies is considered to be incomplete. Such antibodies do not reduce shedding of virus after HRSV and BRSV infection. RSV is often transmitted by contact with nasal secretions but may also be transmitted by aerosols. Seroprevalence of HRSV and BRSV among adult human beings and cattle is over 70% and is always higher than it is among younger individuals. Both human beings and cattle of all ages may be reinfected with RSV. During BRSV reinfections, signs of respiratory tract disease and shedding of virus are seldom observed whereas these are common during HRSV reinfections. Persistent HRSV and BRSV infections in human beings and cattle have been suggested but have not so far been reported.

A glycoprotein E deletion mutant of bovine herpesvirus 1 is avirulent in calves

A marker vaccine elicits an antibody response in the host that can be distinguished from the antibody response induced by a wild-type strain. To obtain a bovine herpesvirus 1 (BHV-1) marker vaccine, we constructed a glycoprotein E (gE) deletion mutant. This was obtained by removing the complete gE coding region from the BHV-1 genome. To attenuate the gE deletion mutant further, we also introduced a small deletion in the thymidine kinase (TK) gene. We selected three mutants: the gE deletion mutant, a TK deletion mutant and a gE/TK double deletion mutant, and examined their virulence and immunogenicity in calves. After intranasal inoculation, the TK deletion mutant showed some residual virulence, whereas the gE and gE/TK deletion mutants were avirulent. The calves inoculated with the deletion mutants were protected against disease after challenge exposure and shed significantly less virus than control calves. Deleting the gE gene, therefore, has little effect on the immunogenicity of BHV-1, but is sufficient to reduce the virulence of BHV-1 in calves. These findings led us to conclude that the gE deletion mutant is a good candidate for a modified live BHV-1 marker vaccine.

Virulence, immunogenicity and reactivation of bovine herpesvirus 1 mutants with a deletion in the gC, gG, gI, gE, or in both the gI and gE gene

Within the framework of developing a marker vaccine against bovine herpesvirus 1 (BHV1), several mutants with deletions in non-essential glycoprotein genes were constructed. Glycoprotein gC, gG, gI and gE single deletion mutants, a gI/gE double deletion mutant and a gE frame-shift mutant were made. The virulence and immunogenicity of these mutants were evaluated in specific-pathogen-free calves. Except for the gC deletion mutant, all mutants were significantly less virulent than the parental wild-type (wt) BHV1 strain Lam. The virulence of the gI and the gI-/gE- mutants was almost completely reduced. Upon challenge infection, the calves of the control group became severely ill, whereas all other calves remained healthy. The reduction of the virus shedding after challenge infection was related to the virulence of the strain of primary inoculation. Virus shedding was almost completely reduced in calves first inoculated with Lam-wt or with gC- and the least reduced in calves inoculated with gI- or gI-/gE-. Six weeks after challenge, all calves were treated with dexamethasone to study whether mutant or challenge virus or both could be reactivated. The gC- and the gG- mutants were reactivated, whereas none of the other mutants were reisolated. Reactivation of challenge virus was reduced in all calves inoculated with mutant viruses. The gC deletion mutant was too virulent and the gI and the gI/gE deletion mutants were the least immunogenic, but based on residual virulence and immunogenicity, both the gG and the gE deletion mutants are candidates for incorporation in live BHV1 vaccines. However, it also depends on the kinetics of the anti-gG and anti-gE antibody response after wild-type virus infection, whether these deletion mutants are really suitable to be incorporated in a marker vaccine.

Advances in the development and evaluation of bovine herpesvirus 1 vaccines.

This review deals with conventional and modern bovine herpesvirus 1 (BHV1) vaccines. Conventional vaccines are widely used to prevent clinical signs of infectious bovine rhinotracheitis. The use of conventional vaccines, however, does not appear to have resulted in reduction of the prevalence of infection. Novel BHV1 marker vaccines comprise either mutants with a deletion in one of the non-essential genes, or subunit vaccines that contain one or more glycoproteins. These marker vaccines can be used in conjunction with companion diagnostic tests to differentiate between infected and vaccinated cattle. Their efficacy has been evaluated in vaccination-challenge experiments, transmission experiments and in field trials. The results demonstrate that the marker vaccines can contribute to the eventual eradication of BHV1. However, there remains room for improvement of BHV1 marker vaccines.

Bovine herpesvirus 1 in semen of bulls and the risk of transmission: A brief review

Summary Outbreaks of bovine herpesvirus 1 (BHV1) infections in artificial insemination centres can run a clinical or sub‐clinical course. Clinical signs vary from mild to severe balanoposthitis and may be associated with a decrease in semen quality. After intranasal as well as intrapreputial infection, BHV1 can replicate in the preputial and penile mucosae, although the pattern of virus shedding in semen differs considerably per individual bull. Beyond the primary phase of a genital infection, BHV1 remains latent in sacral ganglia, and consequently a protracted course of intermittent virus excretion may follow. The seminal plasma rather than the sperm cells contains the BHV1. Diluting the semen before inoculating cell cultures appears to be the best method to neutralize its toxic activity and to achieve optimal virus isolation results. Detection of BHVI in semen by polymerase chain reaction seems to be more sensitive than virus isolation. Not each extended semen straw contains virus when the virus titre i...