Bovine herpesvirus-1 (BoHV-1) is a pathogen of cattle, which most commonly affects the upper respiratory tract to cause infectious bovine rhinotracheitis (IBR). It can also spread systemically to cause fatalities in calves and abortion in pregnant cattle. The virally encoded mechanisms of this systemic spread are poorly understood and therefore have been addressed by comparing isolates from the respiratory form of disease with isolates that have previously demonstrated systemic spread. A survey of 400 bovine abortions in Scotland from 2007-2009 demonstrated a BoHV-1 prevalence of 2.5%. It also demonstrated the importance of real-time PCR as a diagnostic technique when analysing samples from natural cases. The study of BoHV-1 distribution in the placenta and foetal tissue provided support for a haematogenous route of viral spread. Whole genome sequencing of 11 BoHV-1 isolates using Illumina Solexa technology was completed and added significantly to the sequencing data of BoHV-1. In terms of identifying genetic variation between isolates causing respiratory infection and those causing systemic infection, no differences were observed by SNP or phylogenetic analysis. However, there were significant differences in the extent of variation between essential and non-essential genes, which may reflect the evolution of BoHV-1. An in vivo challenge of the natural host to compare two isolates representing the respiratory and systemic forms of infection showed differences in clinical presentation, histopathological analysis, viral distribution and viral transcript expression, measured throughout the infection period. In particular, it was noted that a more severe ocular infection, rather than respiratory based infection was caused by infection with the ‘systemic’ isolate. Differences in the tropism of the virus were observed early in the infection with the ‘systemic’ isolate showing more association with the nasal mucosa than the trachea. The tonsils demonstrated different responses to the virus and differences in viral transcript expression. However, this may simply represent different stages of virus infection. Both isolates demonstrated spread to the brain at day 10 post infection. In vitro methods were used to study the differences in transcript expression in more detail. In a bovine turbinate cell infection faster replication of the respiratory isolate was observed by a significantly faster development of cytopathic effect. This was also reflected in the higher gene expression levels of the respiratory isolate in the first 12 hours of infection. More isolates were studied to investigate whether these differences were consistent, or as suggested by the sequencing, random differences between isolates. Six isolates were used to infect bovine lung slices. Differences in transcript expression were minimal between the two isolate groups. Immunofluorescence did not provide the sensitivity to detect virus in all samples where PCR showed replication. This compromised the study of co-localization but did show promise as a model to study the tropism of respiratory viruses. Overall, this work has showed that systemic spread of BoHV-1 does not appear to be controlled by virally encoded mechanisms. The in vivo experimental infection suggested host factors may play a large part. Further work is also needed to consider any differences that may exist between reactivated virus and the original infecting isolate.

Dissertation (Ph.D.)--Auburn University, 2007. / Abstract. Vita. Includes bibliographic references (p.281-340).

Bovine Infected Cell Protein 0 (bICP0) is an immediate-early protein encoded by Bovine Herpesvirus-1 that modulates host immune response, activates transcription for all viral promoters, and causes ubiquitin-dependent degradation of proteins. Presented herein is a bICP0 protein-protein interaction (PPI) profile, consisting of 98 cellular and 15 viral proteins, generated through co-immunoprecipitation of bICP0 and its binding partners. The PPI profile was analyzed computationally to identify potential sites of interaction with bICP0 and any cellular pathways that may be influenced by bICP0. Some interactors fall in conjunction with bICP0’s known roles during infection, and others are consistent with known associations of bICP0 homologs. However, some proteins in the PPI profile are involved in apoptosis signaling and mRNA spicing—processes both significant during viral infection and novel to the known functions of bICP0 and its homologs. The interaction and co-localization of some of these proteins with bICP0 was further examined.

O presente trabalho avaliou índices reprodutivos e características de desempenho em fêmeas bovinas de corte, infectadas e não infectadas pelo HVB-1, criadas sob manejo extensivo, em uma fazenda na região norte do Estado de São Paulo, Brasil. Animais das raças Gir, Guzerá, Nelore e Caracu foram avaliados no início da estação de monta e a ocorrência de touros e fêmeas reagentes ao HVB-1 pelo teste ELISA foi, respectivamente, 92,5% (37/40) e 54,2% (386/712), sendo o número de touros reagentes maior do que o de fêmeas (p < 0,0001). Foi verificado um aumento na proporção de fêmeas reagentes nas diferentes faixas etárias: 23,2% (32/138) de 2 a 3 anos; 45,2% (57/126) de 3 a 4 anos; 54,6% (59/108) de 4 a 5 anos e 70,0% (238/340) para &#61619; 5 anos (p < 0,0001). O número de matrizes que soroconverteram no período de um ano foi igual a 10,3% (58/561). O HVB-1 não reduziu o índice de prenhez de matrizes reagentes - 80,3% (310/386) e não reagentes - 74,5% (243/326) (p > 0,05) e nem a taxa de parição de matrizes reagentes - 97,7% (300/307) e não reagentes - 93,8% (225/240) (p < 0,05). O índice de prenhez e parição de fêmeas reagentes não diferiu segundo a raça, soroconversão, grupo genético do rebanho Nelore e faixa etária (p > 0,05). O coeficiente de natimortalidade de matrizes reagentes ao HVB-1 - 1,3% (4/300) não diferiu da encontrada para as não reagentes - 2,2% (5/225) (p > 0,05) e não foram observados efeitos de raça, soroconversão, grupo genético do rebanho Nelore e faixa etária (p > 0,05). O HVB-1 não afetou a média de algumas características de desempenho de fêmeas reagentes e não reagentes (p > 0,05), respectivamente, como ganho de peso médio diário durante a estação de monta (459,90 &#61617; 2,82 g e 466,63 &#61617; 2,87 g), condição corporal na entrada da estação de monta (6,89 &#61617; 0,08 e 6,99 &#61617; 0,08), condição corporal na saída da estação de monta (7,73 &#61617; 0,06 e 7,71 &#61617; 0,06) e peso à parição (419,17 &#61617; 3,34 kg e 425,97 &#61617; 3,22 kg). Concluiu-se que matrizes de corte infectadas pelo HVB-1 e não vacinadas, criadas sob condições adequadas de manejo zootécnico, apresentaram bons índices de prenhez, parição e natalidade, independente da raça, grupo genético e faixa etária. / This work evaluated reproductive rates and performance traits in beef cattle females, infected and non infected by BHV-1, bred under extensive management conditions, in a farm in the northern region of São Paulo State, Brazil. Gir, Guzerá, Nelore and Caracu purebred animals were monitorated at the beginning of the breeding season and the occurrence of bulls and females reagent to BHV-1 by ELISA test was, respectively, 92.5% (37/40) and 54.2% (386/712), with positive cases among bulls higher than among females (p < 0.0001). There was an increase in the proportion of reagent females in the different ages analyzed: 23.2% (32/138) from 2 to 3 years old; 45.2% (57/126) from 3 to 4 years old; 54.6% (59/108) from 4 to 5 years old and 70.0% (238/340) above 5 years old (p < 0.0001). The rate of females that seroconverted was 10,3% (58/561). BHV-1 did not interfere in the pregnancy rates of both reagent - 80.3% (310/386) and non-reagent - 74.5% (243/326) females (p > 0.05). It did not reduce the parturition rate of both reagent - 97.7% (300/307) and non-reagent - 93.8% (225/240) females, either (p < 0.05). The pregnancy and parturition rates of reagent females did not differ according to breed, seroconversion, Nelore herd genetic group and age (p > 0.05). Total rate of stillbirths in BHV-1 reagent females - 1.3% (4/300) did not differ from that found in non-reagent females - 2.2%(5/225) (p > 0.05) and did not differ acording to breed, seroconversion, Nelore herd genetic group and age (p > 0.05). BHV-1 did not affect performance traits for reagent and non-reagent females (p > 0.05), respectivelly, to daily weight gain during the breeding season (459.90 &#61617; 2.82 g and 466.63 &#61617; 2.87 g), body condition score at the beginning of the breeding season (6.89 &#61617; 0.08 and 6.99 &#61617; 0.08), body condition score at the end of the breeding season (7.73 &#61617; 0.06 and 7.71 &#61617; 0.06), weight at parturition (419.17 &#61617; 3.34 kg and 425.97 &#61617; 3.22 kg). It was concluded that non-vaccinated beef cattle females infected by BHV-1 and bred under adequate management conditions, presented good pregnancy, parturition and birth rates, no matter the breed, genetic group and age.

O presente trabalho avaliou índices reprodutivos e características de desempenho em fêmeas bovinas de corte, infectadas e não infectadas pelo HVB-1, criadas sob manejo extensivo, em uma fazenda na região norte do Estado de São Paulo, Brasil. Animais das raças Gir, Guzerá, Nelore e Caracu foram avaliados no início da estação de monta e a ocorrência de touros e fêmeas reagentes ao HVB-1 pelo teste ELISA foi, respectivamente, 92,5% (37/40) e 54,2% (386/712), sendo o número de touros reagentes maior do que o de fêmeas (p < 0,0001). Foi verificado um aumento na proporção de fêmeas reagentes nas diferentes faixas etárias: 23,2% (32/138) de 2 a 3 anos; 45,2% (57/126) de 3 a 4 anos; 54,6% (59/108) de 4 a 5 anos e 70,0% (238/340) para &#61619; 5 anos (p < 0,0001). O número de matrizes que soroconverteram no período de um ano foi igual a 10,3% (58/561). O HVB-1 não reduziu o índice de prenhez de matrizes reagentes - 80,3% (310/386) e não reagentes - 74,5% (243/326) (p > 0,05) e nem a taxa de parição de matrizes reagentes - 97,7% (300/307) e não reagentes - 93,8% (225/240) (p < 0,05). O índice de prenhez e parição de fêmeas reagentes não diferiu segundo a raça, soroconversão, grupo genético do rebanho Nelore e faixa etária (p > 0,05). O coeficiente de natimortalidade de matrizes reagentes ao HVB-1 - 1,3% (4/300) não diferiu da encontrada para as não reagentes - 2,2% (5/225) (p > 0,05) e não foram observados efeitos de raça, soroconversão, grupo genético do rebanho Nelore e faixa etária (p > 0,05). O HVB-1 não afetou a média de algumas características de desempenho de fêmeas reagentes e não reagentes (p > 0,05), respectivamente, como ganho de peso médio diário durante a estação de monta (459,90 &#61617; 2,82 g e 466,63 &#61617; 2,87 g), condição corporal na entrada da estação de monta (6,89 &#61617; 0,08 e 6,99 &#61617; 0,08), condição corporal na saída da estação de monta (7,73 &#61617; 0,06 e 7,71 &#61617; 0,06) e peso à parição (419,17 &#61617; 3,34 kg e 425,97 &#61617; 3,22 kg). Concluiu-se que matrizes de corte infectadas pelo HVB-1 e não vacinadas, criadas sob condições adequadas de manejo zootécnico, apresentaram bons índices de prenhez, parição e natalidade, independente da raça, grupo genético e faixa etária. / This work evaluated reproductive rates and performance traits in beef cattle females, infected and non infected by BHV-1, bred under extensive management conditions, in a farm in the northern region of São Paulo State, Brazil. Gir, Guzerá, Nelore and Caracu purebred animals were monitorated at the beginning of the breeding season and the occurrence of bulls and females reagent to BHV-1 by ELISA test was, respectively, 92.5% (37/40) and 54.2% (386/712), with positive cases among bulls higher than among females (p < 0.0001). There was an increase in the proportion of reagent females in the different ages analyzed: 23.2% (32/138) from 2 to 3 years old; 45.2% (57/126) from 3 to 4 years old; 54.6% (59/108) from 4 to 5 years old and 70.0% (238/340) above 5 years old (p < 0.0001). The rate of females that seroconverted was 10,3% (58/561). BHV-1 did not interfere in the pregnancy rates of both reagent - 80.3% (310/386) and non-reagent - 74.5% (243/326) females (p > 0.05). It did not reduce the parturition rate of both reagent - 97.7% (300/307) and non-reagent - 93.8% (225/240) females, either (p < 0.05). The pregnancy and parturition rates of reagent females did not differ according to breed, seroconversion, Nelore herd genetic group and age (p > 0.05). Total rate of stillbirths in BHV-1 reagent females - 1.3% (4/300) did not differ from that found in non-reagent females - 2.2%(5/225) (p > 0.05) and did not differ acording to breed, seroconversion, Nelore herd genetic group and age (p > 0.05). BHV-1 did not affect performance traits for reagent and non-reagent females (p > 0.05), respectivelly, to daily weight gain during the breeding season (459.90 &#61617; 2.82 g and 466.63 &#61617; 2.87 g), body condition score at the beginning of the breeding season (6.89 &#61617; 0.08 and 6.99 &#61617; 0.08), body condition score at the end of the breeding season (7.73 &#61617; 0.06 and 7.71 &#61617; 0.06), weight at parturition (419.17 &#61617; 3.34 kg and 425.97 &#61617; 3.22 kg). It was concluded that non-vaccinated beef cattle females infected by BHV-1 and bred under adequate management conditions, presented good pregnancy, parturition and birth rates, no matter the breed, genetic group and age.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-11-13T12:15:03Z No. of bitstreams: 1 texto completo.pdf: 1654335 bytes, checksum: bce0b05d7832cce3f6841e6c11b57384 (MD5) / Made available in DSpace on 2018-11-13T12:15:03Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1654335 bytes, checksum: bce0b05d7832cce3f6841e6c11b57384 (MD5) Previous issue date: 2018-07-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O bovine herpesvirus 1 (BoHV-1) é de fácil disseminação, difícil controle e está amplamente disseminado nos rebanhos bovinos no mundo. Essa infecção viral é responsável por prejuízos à pecuária principalmente quanto aos aspectos reprodutivos. Estudos prévios envolvendo infecção experimental com o BoHV-1 em um sistema de produção in vitro de embriões reportou que a presença do vírus afetou o desenvolvimento embrionário. Neste trabalho, foi avaliada a interferência do BoHV-1 sobre a maturação in vitro de complexos cumulus-ovócito (COCs) e ovócitos desnudados com células do cumulus em suspensão (DOs). Foram coletados sangue e os ovários de fêmeas bovinas sabidamente não vacinadas contra o BoHV-1. Após o teste de soroneutralização, os animais soropositivos foram classificados de acordo com a sua titulação de anticorpos. Os ovócitos recuperados após aspiração folicular foram divididos em COCs e DOs e avaliados através da sua capacidade de maturação nuclear associado a detecção viral pela microscopia confocal de varredura a laser. Dois experimentos foram realizados: (I) maturação após infecção in vitro de COCs e DOs de animais soronegativos; (II) maturação in vitro de COCs e DOs de animais soropositivos. No experimento I, diferença (P<0,01) foi observada entre as taxas de maturação do grupo COCs controle (78,2%) e os grupos infectados dos COCs (43,6%). No experimento II verificou-se diferença (P<0,01) na taxa de maturação entre os animais de titulação de anticorpos ≥ 16 (56,9%) e o grupo controle (79,4%). Os ensaios de imunofluorescência permitiram a identificação do BoHV-1 nos COCs e DOs. Diante disso, conclui-se que o BoHV-1 foi capaz de afetar o processo de maturação in vitro tanto nas condições de infecção in vitro e em condições naturais de infecção. / Bovine herpesvirus 1 (BoHV-1) disseminates easily, is difficult to control, and is widely spread in cattle herds worldwide. BoHV-1 causes a broad range of losses to the cattle industry, mainly concerning reproduction. Previous studies involving experimental infection of BoHV-1 in an in vitro embryo production system have reported impairment of embryonic development by BoHV-1. In this study, we evaluated the interference of BoHV-1 in the in vitro maturation system of cumulus- oocyte complexes (COCs) and denuded oocytes (DOs) cultured with a cumulus cell suspension. Blood samples and ovaries were collected from slaughterhouse cows unvaccinated against BoHV-1. Using virus neutralization assays, the seropositive animals were classified according to their antibody titers. The oocytes were recovered by follicular aspiration and divided into two groups, COCs and DOs, which were evaluated for their nuclear maturation capacity using immunofluorescence assays by laser scanning confocal microscopy. Two experiments were carried out: (I) in vitro maturation of COCs and DOs after artificial infection of seronegative animals and (II) in vitro maturation of COCs and DOs of seropositive animals. In experiment I, a difference (P <0.01) was observed between the maturation rates of the control group COCs (78.2%) and the infected COCs (43.6%). In experiment II, there was a difference (P <0.01) in the maturation rate between animals with antibody titers ≥ 16 (56.9%) and the control group (79.4%). Immunofluorescence assays identified BoHV-1 in the COCs and DOs. Therefore, it was concluded that BoHV-1 affects the in vitro maturation process in both in vitro and natural infections.

Bovine Respiratory Disease Complex is a multifactorial disease affecting cattle worldwide resulting in high mortality and morbidity rates in the cattle farming industry. This complex is caused by multiple viral and bacterial pathogens such as Bovine Herpesvirus-1, Bovine Respiratory Syncytial Virus, Mannheimia haemolytica, and Pasteurella multocida; two of the main contributors to the initiation of this disease are Bovine Herpesvirus-1 and the bacteria, Mannheimia haemolytica. Together, these microbes co-infect immunocompromised cattle during times of increased stress and induce a severe pneumonic response along with other health complications. Research has been primarily focused on these microorganisms individually or their effect on the host, however there is a need to study them together due to the increased mortality rate associated with co-infections. In this study, we used Bovine Herpesvirus-1, Mannheimia haemolytica, Pasteurella multocida, and Bovine Respiratory Syncytial Virus to co-infect bovine tissue cultures to determine how they affect each other.

Bovine herpesvirus-1 (BoHV-1) is an important pathogen of cattle associated with respiratory and reproductive disease and is the most common viral agent implicated in the bovine respiratory disease complex (BRDC). BRDC is an economically significant multifactorial disease of feedlot cattle estimated to cost Australian feedlot producers $AU60 million/year in lost production, therapeutics and disease management. Worldwide BRDC is attributed to cost $US2 billion to cattle industries. In an effort to limit the associated economic costs and enhance animal health and welfare of feedlot cattle, the concerted use of vaccination and diseased animal management are practiced. Numerous vaccines are available in North America and Canada however, in Australia, feedlot producers are reliant on three vaccines. These vaccines target either the bacterial or viral agents of the BRDC and encompass antibody, subunit and attenuated live BoHV-1 preparations. Live attenuated vaccines are developed by numerous methods including, deletion or disruption of certain genes. The development of an attenuated live virus vaccine was traditionally a laborious task requiring numerous rounds of in vitro purification. Contrastingly, technological advances introduced this decade, allowing the stable maintenance of the complete herpesvirus genome in bacteria as a bacterial artificial chromosome (BAC), has advanced herpes virology exponentially in that investigation and manipulation of the herpesvirus genome can be conducted independent of a cell culture system. With respect to BRDC and the generation of vaccines to combat the disease, the tools to fully utilise the potential of BoHV-1 as a live vaccine vector are now routine. It is now possible to vii construct BoHV-1 as a delivery vector by inserting appropriate antigens of those bacterial and viral pathogens implicated in the BRDC into a BAC maintained BoHV-1 genome. However, there is a significant lack of genetic information regarding BoHV-1 and inserting several antigenic sequences would expand the genome of BoHV-1 inducing non-viability. Therefore, to further develop BoHV-1 as a vaccine vector, a study was conducted to identify the essential and nonessential genes required for the in vitro viability of BoHV-1. Identifying the essential and nonessential genes will establish which genes may be preferentially deleted or replaced with exogenous antigenic sequences in a BoHV-1 derived vaccine vector. To define the requirement of genes encoded by BoHV-1, random-insertion mutagenesis utilising a Tn5 transposition system and targeted gene deletion catalysed by GET recombination was employed to construct gene disruption and gene deletion libraries, respectively, of an infectious clone of BoHV-1. Transposon insertion position and confirmation of gene deletion was determined by direct sequencing. with the essential or nonessential requirement of either transposed or deleted open reading frames (ORFs) assessed by transfection of respective BoHV- 1 BAC DNA into host cells. Of the 73 recognised ORFs encoded by the BoHV-1 genome, 33 were determined to be essential and 36 to be nonessential for virus viability in cell culture with the requirement of the two dual copy ORFs inconclusive. The majority of ORFs were shown to conform to the in vitro requirements of BoHV-1 homologues encoded by Human herpesvirus 1. However, ORFs encoding for glycoprotein K (UL53), regulatory, membrane, tegument and capsid proteins (UL54, UL49.5, UL49, UL35, UL20, UL16 and UL7) were shown to differ in requirement when compared to Human herpesvirus-1 encoded homologues. Further analysis of clones encompassing restriction digestion profiling, one-step growth and replication kinetic analysis defined the genetic constitution and replicative capacity of the mutant clones. Thirty-three individual ORFs of the 36 defined nonessential ORF were identified as being amenable to deletion without causing significant replicative detriment to a potential BoHV-1 vaccine vector. This study has provided the foundational information required for the future development of BoHV-1 as a multivalent vaccine vector for the protection of feedlot cattle from BRDC. Furthermore, the genetic information generated in this study contributes to the general knowledge of the prototype ruminant herpesvirus, BoHV-1, and contributes to the comparative study of gene function between the large and diverse family that is Herpesviridae.

Bovine herpesvirus-1 (BoHV-1) is an important pathogen of cattle associated with respiratory and reproductive disease and is the most common viral agent implicated in the bovine respiratory disease complex (BRDC). BRDC is an economically significant multifactorial disease of feedlot cattle estimated to cost Australian feedlot producers $AU60 million/year in lost production, therapeutics and disease management. Worldwide BRDC is attributed to cost $US2 billion to cattle industries. In an effort to limit the associated economic costs and enhance animal health and welfare of feedlot cattle, the concerted use of vaccination and diseased animal management are practiced. Numerous vaccines are available in North America and Canada however, in Australia, feedlot producers are reliant on three vaccines. These vaccines target either the bacterial or viral agents of the BRDC and encompass antibody, subunit and attenuated live BoHV-1 preparations. Live attenuated vaccines are developed by numerous methods including, deletion or disruption of certain genes. The development of an attenuated live virus vaccine was traditionally a laborious task requiring numerous rounds of in vitro purification. Contrastingly, technological advances introduced this decade, allowing the stable maintenance of the complete herpesvirus genome in bacteria as a bacterial artificial chromosome (BAC), has advanced herpes virology exponentially in that investigation and manipulation of the herpesvirus genome can be conducted independent of a cell culture system. With respect to BRDC and the generation of vaccines to combat the disease, the tools to fully utilise the potential of BoHV-1 as a live vaccine vector are now routine. It is now possible to vii construct BoHV-1 as a delivery vector by inserting appropriate antigens of those bacterial and viral pathogens implicated in the BRDC into a BAC maintained BoHV-1 genome. However, there is a significant lack of genetic information regarding BoHV-1 and inserting several antigenic sequences would expand the genome of BoHV-1 inducing non-viability. Therefore, to further develop BoHV-1 as a vaccine vector, a study was conducted to identify the essential and nonessential genes required for the in vitro viability of BoHV-1. Identifying the essential and nonessential genes will establish which genes may be preferentially deleted or replaced with exogenous antigenic sequences in a BoHV-1 derived vaccine vector. To define the requirement of genes encoded by BoHV-1, random-insertion mutagenesis utilising a Tn5 transposition system and targeted gene deletion catalysed by GET recombination was employed to construct gene disruption and gene deletion libraries, respectively, of an infectious clone of BoHV-1. Transposon insertion position and confirmation of gene deletion was determined by direct sequencing. with the essential or nonessential requirement of either transposed or deleted open reading frames (ORFs) assessed by transfection of respective BoHV- 1 BAC DNA into host cells. Of the 73 recognised ORFs encoded by the BoHV-1 genome, 33 were determined to be essential and 36 to be nonessential for virus viability in cell culture with the requirement of the two dual copy ORFs inconclusive. The majority of ORFs were shown to conform to the in vitro requirements of BoHV-1 homologues encoded by Human herpesvirus 1. However, ORFs encoding for glycoprotein K (UL53), regulatory, membrane, tegument and capsid proteins (UL54, UL49.5, UL49, UL35, UL20, UL16 and UL7) were shown to differ in requirement when compared to Human herpesvirus-1 encoded homologues. Further analysis of clones encompassing restriction digestion profiling, one-step growth and replication kinetic analysis defined the genetic constitution and replicative capacity of the mutant clones. Thirty-three individual ORFs of the 36 defined nonessential ORF were identified as being amenable to deletion without causing significant replicative detriment to a potential BoHV-1 vaccine vector. This study has provided the foundational information required for the future development of BoHV-1 as a multivalent vaccine vector for the protection of feedlot cattle from BRDC. Furthermore, the genetic information generated in this study contributes to the general knowledge of the prototype ruminant herpesvirus, BoHV-1, and contributes to the comparative study of gene function between the large and diverse family that is Herpesviridae.