TLR activation, immune response and viral protection elicited in cattle by a commercial vaccine against Bovine Herpesvirus-1.

The innate and acquired immune response induced by a commercial inactivated vaccine against Bovine Herpesvirus-1 (BoHV-1) and protection conferred against the virus were analyzed in cattle. Vaccination induced high levels of BoHV-1 antibodies at 30, 60, and 90 days post-vaccination (dpv). IgG1 and IgG2 isotypes were detected at 90 dpv, as well as virus-neutralizing antibodies. An increase of anti-BoHV-1 IgG1 in nasal swabs was detected 6 days post-challenge in vaccinated animals. After viral challenge, lower virus excretion and lower clinical score were observed in vaccinated as compared to unvaccinated animals, as well as BoHV-1-specific proliferation of lymphocytes and production of IFNγ, TNFα, and IL-4. Downregulation of the expression of endosome Toll-like receptors 8-9 was detected after booster vaccination. This is the first thorough study of the immunity generated by a commercial vaccine against BoHV-1 in cattle.

Leptospirosis, bovine viral diarrhea and infectious bovine rhinotracheitis: prevalence in Colombian cattle and buffaloes

One of the limiting factors of productive efficiency in cattle and buffalo herds is related to the high prevalence of infectious diseases which affect reproduction. The aim of this study was to determine the prevalence of anti-Leptospiraantibodies for bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BoHV-1) in bovine and buffalo herds in Colombia. Blood serum samples were collected from 1100 buffaloes and 1000 cattle. The ELISA technique was used to detect antibodies against BVDV and BoHV-1, and the microscopic agglutination technique to detect anti-Leptospiraantibodies. The prevalence of anti-Leptospiraantibodies and of BVDV and BoHV-1 in bovine samples was observed in 16, 39.7, and 65% of animals, respectively, while the positivity in samples for the same antibodies in buffalos was detected in 18.7, 27.5 and 51.5%, respectively. Exposure of cattle and buffaloes to BoHV-1 was positively associated with age, higher prevalence rates were observed in older ages. Seropositivity of cattle for BVDV and BoHV-1 was higher in male animals. Activities such as embryo transfer, milking, and needle reuses, as well as the presence of cats and rodents are factors which favor positivity of the herd for BVDV and BoHV-1.(AU)

Economic impacts of Bovine alphaherpesvirus 1 infection in Brazil: Meta-analysis based on epidemiological indicators / Impactos econômicos da infecção por Bovine alphaherpesvirus 1 no Brasil: Metanálise a partir de indicadores epidemiológicos

This study aimed to conduct a systematic review to estimate the economic impact of Bovine alphaherpesvirus 1 (BoAHV1) infection in Brazil using epidemiological indicators through a meta-analysis. Specific descriptors were used to retrieve studies from the Scopus, PubMed, Biblioteca Digital Brasileira de Teses e Dissertações, and Catálogo de Teses e Dissertações da Capes databases, selecting those that met the inclusion criteria established between the years 2000 and 2020. The selected studies were subjected to descriptive statistical analysis using prevalence data as the primary outcome with a 95% confidence interval (CI) with a meta-analysis of random effects and measures of heterogeneity, significance, magnitude of the effect, and measurement of publication bias. Abortion costs and estimates were calculated based on the prevalence of BoAHV1 infection in Brazil and the characteristics of the agent as viral subtypes that cause abortion, period of occurrence, average pregnancy rate, and morbidity applied to susceptible animals and animal replacement values. The results were obtained from 49 studies included for meta-analysis where a prevalence of BoAHV1 infection of 54.12% (95% CI: 49.07% - 59.26%) in the bovine population and 88.53% (95% CI: 82.97%­92.43%) was present. From the structured formula, it is estimated that 258,779 bovine abortions occur, which causes a total loss, based only on the occurrence of abortion, of US $ 48,402,244.00 to the country. It is noteworthy that because of the losses caused, strict control and eradication measures need to be implemented based on the elaboration of normative instruction that includes health education measures, vaccination, tests for traffic, and animal trade so that BoAHV1 infections do not continue to negatively impact national producers economically, productively, and socially.(AU)

Objetivou-se com o presente estudo realizar uma revisão sistemática para estimar os impactos econômicos da infecção por Alfaherpesvirus Bovino 1 no Brasil a partir dos indicadores epidemiológicos por meio de uma metanálise. Foram usados descritores específicos para a recuperação de estudos a partir das bases Scopus, Pubmed, Biblioteca Digital Brasileira de Teses e Dissertações e do Catálogo de Teses e Dissertações da Capes, selecionando-se aqueles que se enquadraram nos critérios de inclusão estabelecidos entre os anos 2000 e 2020. Os estudos selecionados foram submetidos à análise estatística descritiva utilizando-se os dados de prevalência como desfecho primário com intervalo de confiança de 95%, sendo realizado metanálise de efeito randômico e avaliadas as medidas de heterogeneidade, significância, magnitude do efeito e aferição do viés de publicação. Os custos e estimativas de aborto foram calculados com base na prevalência encontrada para BoAHV1 no Brasil e as características do agente como subtipos virais que provocam o aborto, período de ocorrência, taxa média de prenhez e morbidade aplicados aos animais suscetíveis e aos valores de reposição animal. Os resultados foram obtidos a partir de 49 estudos incluídos para metanálise, onde se verificou uma prevalência de BoAHV1 de 54,12% (IC95%: 49,07% ­ 59,26%) na população bovina e 88,53% (IC95%: 82,97% ­ 92,43%) para os rebanhos. A partir da fórmula estruturada, se estima que ocorram 258.779 abortos bovinos, que totalizam um prejuízo, apenas baseado na ocorrência do aborto, de US$ 48.402.244,00 ao país. Destaca-se que em virtude dos prejuízos ocasionados, medidas de controle e erradicação mais rigorosas precisam ser implementadas a partir da elaboração de uma instrução normativa que contemple medidas de educação sanitária, vacinação, realização de exames para o trânsito e comércio de animais para que as infecções pelo BoAHV1 não continuem a impactar negativamente os produtores nacionais de forma econômica, produtiva e socialmente.(AU)

Distinctive features of bovine alphaherpesvirus types 1 and 5 and the virus-host interactions that might influence clinical outcomes.

Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are two closely related alphaherpesviruses. BoHV-1 causes several syndromes in cattle, including respiratory disease and sporadic cases of encephalitis, whereas BoHV-5 is responsible for meningoencephalitis in calves. Although both viruses are neurotropic, they differ in their neuropathogenic potential. This review summarizes the findings on the specific mechanisms and pathways known to modulate the pathogenesis of BoHV-1 and BoHV-5, particularly in relation to respiratory and neurological syndromes, which characterize BoHV-1 and BoHV-5 infections, respectively.

Proteomic and phosphoproteomic analyses reveal several events involved in the early stages of bovine herpesvirus 1 infection.

Herpesviruses are predicted to express more than 80 proteins during their infection cycle. The proteins synthesized by the immediate early genes and early genes target signaling pathways in host cells that are essential for the successful initiation of a productive infection and for latency. In this study, proteomic and phosphoproteomic tools showed the occurrence of changes in Madin-Darby bovine kidney cells at the early stage of the infection by bovine herpesvirus 1 (BoHV-1). Proteins that had already been described in the early stage of infection for other herpesviruses but not for BoHV-1 were found. For example, stathmin phosphorylation at the initial stage of infection is described for the first time. In addition, two proteins that had not been described yet in the early stages of herpesvirus infections in general were ribonuclease/angiogenin inhibitor and Rab GDP dissociation inhibitor beta. The biological processes involved in these cellular responses were repair and replication of DNA, splicing, microtubule dynamics, and inflammatory responses. These results reveal pathways that might be used as targets for designing antiviral molecules against BoHV-1 infection.

Bovine herpesvirus 1 can cross the intact zona pellucida of bovine oocytes after artificial infection.

Bovine herpesvirus 1 (BHV1) is an important bovine pathogen, responsible for respiratory diseases and reproductive problems. This study investigated the penetration capacity of BHV1 into oocytes after co-incubation for either 1 h or 24 h. Immunofluorescence assays in cumulus-oocyte complexes (COCs) and denuded oocytes (without the presence of cumulus cells) were performed and evaluated using confocal laser scanning microscopy. Blood samples and ovaries from BHV1 seronegative cows were used. The oocytes recovered were divided into two groups. Group I comprised COCs (n = 312) and denuded oocytes (n = 296), which were experimentally infected with BHV1 and incubated for 1 h at 38.5°C and 5% CO2. Group II comprised COCs (n = 425) and denuded oocytes (n = 405), which were co-incubated with BHV1 under the same conditions for 24 h. The negative control of these two groups was respectively subjected to the same protocol, except for exposure to BHV1. To our knowledge, this study provides the first evidence of BHV1 detection within COCs and denuded oocytes exhibiting intact zona pellucida when co-incubated with the virus for 24 h. Immunolocalization also confirmed the presence of BHV1 in the cytoplasm of the cumulus cells of all COCs exposed to the virus after both incubation periods. In conclusion, detection of BHV1 inside oocytes has a great meaning for the field of animal reproduction. The detection of BHV1 in different layers of cumulus cells also demonstrates that these cells are sources of viral infection.

Neurological diseases of cattle in the state of Goiás, Brazil (2010-2017) / Doenças neurológicas de bovinos no estado de Goiás (2010-2017)

A retrospective study was conducted on neurological diseases of cattle in the state of Goiás, Brazil, from March 2010 to August 2017. Samples of three veterinary diagnostic laboratories were analyzed. Diagnosis was established in 170 out of 407 cattle with neurological signs. Epidemiological, clinical, and anatomic pathology features of each case were researched in the files. Main disorders included diseases caused by viruses (rabies 29.41%, meningoencephalitis by bovine herpesvirus 15.88%, and malignant catarrhal fever 1.76%), by bacteria (botulism 5.88%, suppurative meningitis 3.53%, encephalic abscesses 2.94%, listeriosis 1.76%, and thrombotic meningoencephalitis 1.76%), of metabolic origin (polioencephalomalacia 17.06%), of indefinite cause (lymphoplasmacytic meningoencephalitis 11.18%, traumatic hemorrhages 3.53%, and multifocal malacia with gliosis 1.18%), congenital (hydrocephaly 1.18% and multiple malformations 0.59%), toxic (urea poisoning 1.18% and insecticide poisoning 0.59%), and parasitic (meningoencephalitis associated with infection by Trypanosoma sp. 0.59%).(AU)

Foi realizado um estudo retrospectivo de doenças neurológicas de bovinos no estado de Goiás durante o período de março de 2010 a agosto de 2017, analisando amostras de três laboratórios de diagnóstico veterinário. De 407 bovinos que apresentaram sinais clínicos neurológicos, o diagnóstico foi estabelecido em 170 casos. Desses casos, foram pesquisadas nas fichas as características epidemiológicas, clínicas e anatomopatológicas. As principais doenças diagnosticadas foram causadas por vírus (raiva 29,41%, meningoencefalite por herpesvírus bovino 15,88% e febre catarral maligna 1,76%), de origem metabólica (polioencefalomalacia 17,06%), por bactérias (botulismo 5,88%, meningite supurativa 3,53%, abscessos encefálicos 2,94%, listeriose 1,76% e meningoencefalite trombótica 1,76%), sem causa definida (meningoencefalite linfoplasmocítica 11,18%, hemorragias traumáticas 3,53% e malacia multifocal com gliose 1,18%), congênitas (hidrocefalia 1,18% e malformações múltiplas 0,59%), tóxicas (intoxicação por ureia 1,18% e intoxicação por inseticida 0,59%), e parasitária (meningoencefalite associada à infecção por Trypanosoma sp. 0,59%).(AU)

Neurological diseases of cattle in the state of Goiás, Brazil (2010-2017) / Doenças neurológicas de bovinos no estado de Goiás (2010-2017)

A retrospective study was conducted on neurological diseases of cattle in the state of Goiás, Brazil, from March 2010 to August 2017. Samples of three veterinary diagnostic laboratories were analyzed. Diagnosis was established in 170 out of 407 cattle with neurological signs. Epidemiological, clinical, and anatomic pathology features of each case were researched in the files. Main disorders included diseases caused by viruses (rabies 29.41%, meningoencephalitis by bovine herpesvirus 15.88%, and malignant catarrhal fever 1.76%), by bacteria (botulism 5.88%, suppurative meningitis 3.53%, encephalic abscesses 2.94%, listeriosis 1.76%, and thrombotic meningoencephalitis 1.76%), of metabolic origin (polioencephalomalacia 17.06%), of indefinite cause (lymphoplasmacytic meningoencephalitis 11.18%, traumatic hemorrhages 3.53%, and multifocal malacia with gliosis 1.18%), congenital (hydrocephaly 1.18% and multiple malformations 0.59%), toxic (urea poisoning 1.18% and insecticide poisoning 0.59%), and parasitic (meningoencephalitis associated with infection by Trypanosoma sp. 0.59%).(AU)

Foi realizado um estudo retrospectivo de doenças neurológicas de bovinos no estado de Goiás durante o período de março de 2010 a agosto de 2017, analisando amostras de três laboratórios de diagnóstico veterinário. De 407 bovinos que apresentaram sinais clínicos neurológicos, o diagnóstico foi estabelecido em 170 casos. Desses casos, foram pesquisadas nas fichas as características epidemiológicas, clínicas e anatomopatológicas. As principais doenças diagnosticadas foram causadas por vírus (raiva 29,41%, meningoencefalite por herpesvírus bovino 15,88% e febre catarral maligna 1,76%), de origem metabólica (polioencefalomalacia 17,06%), por bactérias (botulismo 5,88%, meningite supurativa 3,53%, abscessos encefálicos 2,94%, listeriose 1,76% e meningoencefalite trombótica 1,76%), sem causa definida (meningoencefalite linfoplasmocítica 11,18%, hemorragias traumáticas 3,53% e malacia multifocal com gliose 1,18%), congênitas (hidrocefalia 1,18% e malformações múltiplas 0,59%), tóxicas (intoxicação por ureia 1,18% e intoxicação por inseticida 0,59%), e parasitária (meningoencefalite associada à infecção por Trypanosoma sp. 0,59%).(AU)

Bovine herpesvirus type 5 replication and induction of apoptosis in vitro and in the trigeminal ganglion of experimentally-infected cattle.

Bovine herpesvirus (BoHV) types 1 and 5 are neuroinvasive. Cases of BoHV-1-induced encephalitis are not as frequent as those caused by BoHV-5. In this study, the capability of BoHV-5 to induce apoptosis in cell cultures and in the trigeminal ganglion during acute infection of experimentally-infected cattle was analyzed. Apoptotic changes in cell cultures agree with the ability of the viral strains to replicate in each cell line. Marked differences were observed between the in vitro induction of apoptosis by BoHV-1Cooper and BoHV-5 97/613 strains. Apoptotic neurons were clearly evident in the trigeminal ganglion of BoHV-1-infected calves. For BoHV-5 a fewer number of positive neurons was observed. There is an association between the magnitude of bovine herpesviruses replication and the induction of apoptosis in trigeminal ganglion. These findings suggest that the induction of apoptosis and the innate immune response orchestrate the final outcome of alpha herpesviruses infection of the bovine nervous system.

Outbreak Control and Clinical, Pathological, and Epidemiological Aspects and Molecular Characterization of a Bovine Herpesvirus Type 5 on a Feedlot Farm in São Paulo State.

This paper describes the control, epidemiological, pathological, and molecular aspects of an outbreak of meningoencephalitis in calves due to bovine herpesvirus 5 at a feedlot with 540 animals in São Paulo State, Brazil. The introduction of new animals and contact between the resident animals and the introduced ones were most likely responsible for virus transmission. Bovine herpesvirus 1 vaccine was used, resulting in the efficacy of the outbreak control, although two bovine herpesvirus 1 positive animals, vaccinated and revaccinated, presented meningoencephalitis, thereby characterizing vaccinal failure.

Sequence analysis of the 5' third of glycoprotein C gene of South American bovine herpesviruses 1 and 5.

Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) share high genetic and antigenic similarities, but exhibit marked differences in tissue tropism and neurovirulence. The amino-terminal region of glycoprotein C (gC), which is markedly different in each of the viruses, is involved in virus binding to cellular receptors and in interactions with the immune system. This study investigated the genetic and antigenic differences of the 5' region of the gC (5' gC) gene (amino-terminal) of South American BoHV-1 (n=19) and BoHV-5 (n=25) isolates. Sequence alignments of 374 nucleotides (104 amino acids) revealed mean similarity levels of 97.3 and 94.2% among BoHV-1 gC (gC1), respectively, 96.8 and 95.6% among BoHV-5 gC (gC5), and 62 and 53.3% between gC1 and gC5. Differences included the absence of 40 amino acid residues (27 encompassing predicted linear epitopes) scattered throughout 5' gC1 compared to 5' gC5. Virus neutralizing assays testing BoHV-1 and BoHV-5 antisera against each isolate revealed a high degree of cross-neutralization between the viruses, yet some isolates were neutralized at very low titers by heterologous sera, and a few BoHV-5 isolates reacted weakly with either sera. The virus neutralization differences observed within the same viral species, and more pronounced between BoHV-1 and BoHV-5, likely reflect sequence differences in neutralizing epitopes. These results demonstrate that the 5' gC region is well conserved within each viral species but is divergent between BoHV-1 and BoHV-5, likely contributing to their biological and antigenic differences.

Frequência de anticorpos contra o herpesvírus bovino tipo 1 (bohv-1) em bovinos de corte não vacinados na microrregião de imperatriz

O herpesvírus bovino tipo 1 (BoHV-1), é responsável por manifestações clínicas como a rinotraqueíte infecciosa bovina, abortamentos, conjuntivite, balanopostite e vulvovaginite pustular infecciosa. Esse vírus tem sido responsável por grandes prejuízos produtivos e reprodutivos em diversos rebanhos do país. Deste modo, o objetivo desse trabalho foi estimar a frequência de anticorpos contra o BoHV-1 em fêmeas bovinas de corte, não vacinadas na Microrregião de Imperatriz, Maranhão; além de identificar a faixa etária mais acometida pelo vírus, bem como realizar um estudo dos fatores associados à infecção do vírus e avaliar a técnica ELISA indireto utilizando a soroneutralização (SN) como padrão de referência. O estudo foi realizado em 48 rebanhos de corte, distribuídos em 12 municípios da Microrregião de Imperatriz. As amostras foram coletadas de fêmeas bovinas estratificadas em três faixas etárias, ? 12 meses, entre 12 e 36 meses e ? 36 meses de idade. As amostras foram submetidas a dois testes sorológicos, ELISA indireto e SN. Em cada rebanho, foi aplicado um questionário epidemiológico, com o objetivo de obter informações sobre manejo sanitário e reprodutivo, para o estudo de fatores de risco. A frequência de anticorpos contra o BoHV-1 na Microrregião de Imperatriz foi de 63,23%, os municípios de Açailândia e Buritirana, apresentaram as maiores frequências, ambos com 80,44%, a faixa etária mais acometida, na Microrregião, foi a dos animais com idade ? 36 meses (69,65%). Com base nos resultados obtidos pode-se concluir que a frequência de anticorpos contra o BoHV-1 é alta; entre as faixas etárias mais acometidas foram a dos animais com idade ? 36 meses; foram considerados fatores de risco para a transmissão do vírus, retorno ao cio, reposição de animais oriundos de outros estados/região e a criação de caprinos/ovinos associados com bovinos; a técnica ELISA indireta apresentou concordância moderada quando comparada a técnica de SN, que é a técnica padrão ouro de diagnóstico para BoHV-1

Molecular and in vitro characterization of field isolates of bovine herpesvirus-1.

Bovine Herpesvirus-1 (BoHV-1) is distributed worldwide and is a major pathogen in cattle, being the causal agent of a variety of clinical syndromes. The aim of this study was to isolate and to characterize (molecular and biological characterization) BoHV-1 from 29 immunosuppressed animals. It was possible to obtain 18 isolates, each from a different animal, such as from the respiratory and reproductive tracts. In some cases the cytopathic effect was visible 12 hours post-inoculation, and became characteristic after 36-48 hours. Biological characteristics were evaluated and compared with Iowa and Colorado-1 reference strains, and differences were found in plaque size, virus titer measured by TCID50 and PFU/mL, and one step virus curves. These results showed that some isolates had a highly virulent-like behavior in vitro, compared to the reference strains, with shorter eclipse periods, faster release of virus into the supernatants, and higher burst size and viral titer. There were no differences in glycoprotein expression of BoHV-1 isolates, measured by Western blot on monolayers. Moreover, using restriction endonucleases analysis, most of the viruses were confirmed as BoHV-1.1 and just one of them was confirmed as BoHV-1.2a subtype. These findings suggest that some wild-type BoHV-1 isolates could be useful as seeds to develop new monovalent vaccines.

Fatores de risco associados à infecção pelo Herpesvírus Bovino Tipo 1 em rebanhos bovinos leiteiros da região Amazônica maranhense / Risk factors for Bovine Herpesvirus 1 infection in cattle herds in the maranhense Amazon region, Brazil

O presente estudo teve como objetivo determinar a frequência e os fatores de risco para a infecção pelo Herpesvírus Bovino Tipo 1 (BoHV-1) em fêmeas bovinas leiteiras na região Amazônica maranhense. Amostras de soro de animais não vacinados contra o BoHV-1 e provenientes de 40 propriedades foram submetidas à técnica de ELISA indireto. Em cada propriedade avaliada, aplicou-se ainda um questionário epidemiológico para investigar os fatores de risco que poderiam estar associados à infecção pelo vírus. Das 400 amostras de soro analisadas, 69,25% (n = 277) foram reagentes com a detecção de bovinos sorologicamente positivos em 100% (n = 40) das propriedades. Dentre os fatores de risco avaliados, faixa etária entre 3 a 7 anos, ausência de assistência veterinária e sinais respiratórios apresentaram significância estatística (P < 0,05) associada à infecção pelo BoHV-1. Os resultados do estudo indicam que a frequência de anticorpos contra o BoHV-1 foi elevada na região Amazônica maranhense e que os fatores relacionados às características das propriedades e ao manejo foram associados ao risco de infecção pelo BoHV-1.

RISK FACTORS FOR BOVINE HERPESVIRUS 1 INFECTION IN CATTLE HERDS IN THE MARANHENSE AMAZON REGION, BRAZIL. This study was performed in order to determine the frequency of Bovine Herpesvirus 1 (BoHV-1), and the main factors related to the frequency of the infection in dairy cows not vaccinated against BoHV-1 from the Maranhense Amazon region, Brazil. Serum samples from 40 ranches were submitted to indirect ELISA test for detection of BoHV-1 antibodies. An epidemiological questionnaire was applied for each herd to investigate variables that could be associated with this infection. From 400 serum samples examined, 69.25% (n = 277) in 100% (n = 40) of the herds were positive for ELISA. Variables identified as risk factors such of age range of 3–7 years, absence of veterinary assistance and respiratory signs presented a statistical significance (P < 0.05) associated to BoHV-1 infection. These results indicate that BoHV-1 infection is high in the region studied and that factors related to characteristics of the ranches and to the livestock management practices were associated with the risk of infection by BoHV-1.

Fatores de risco associados à infecção pelo Herpesvírus Bovino Tipo 1 em rebanhos bovinos leiteiros da região Amazônica maranhense / Risk factors for Bovine Herpesvirus 1 infection in cattle herds in the maranhense Amazon region, Brazil

O presente estudo teve como objetivo determinar a frequência e os fatores de risco para a infecção pelo Herpesvírus Bovino Tipo 1 (BoHV-1) em fêmeas bovinas leiteiras na região Amazônica maranhense. Amostras de soro de animais não vacinados contra o BoHV-1 e provenientes de 40 propriedades foram submetidas à técnica de ELISA indireto. Em cada propriedade avaliada, aplicou-se ainda um questionário epidemiológico para investigar os fatores de risco que poderiam estar associados à infecção pelo vírus. Das 400 amostras de soro analisadas, 69,25% (n = 277) foram reagentes com a detecção de bovinos sorologicamente positivos em 100% (n = 40) das propriedades. Dentre os fatores de risco avaliados, faixa etária entre 3 a 7 anos, ausência de assistência veterinária e sinais respiratórios apresentaram significância estatística (P < 0,05) associada à infecção pelo BoHV-1. Os resultados do estudo indicam que a frequência de anticorpos contra o BoHV-1 foi elevada na região Amazônica maranhense e que os fatores relacionados às características das propriedades e ao manejo foram associados ao risco de infecção pelo BoHV-1. (AU)

RISK FACTORS FOR BOVINE HERPESVIRUS 1 INFECTION IN CATTLE HERDS IN THE MARANHENSE AMAZON REGION, BRAZIL. This study was performed in order to determine the frequency of Bovine Herpesvirus 1 (BoHV-1), and the main factors related to the frequency of the infection in dairy cows not vaccinated against BoHV-1 from the Maranhense Amazon region, Brazil. Serum samples from 40 ranches were submitted to indirect ELISA test for detection of BoHV-1 antibodies. An epidemiological questionnaire was applied for each herd to investigate variables that could be associated with this infection. From 400 serum samples examined, 69.25% (n = 277) in 100% (n = 40) of the herds were positive for ELISA. Variables identified as risk factors such of age range of 3–7 years, absence of veterinary assistance and respiratory signs presented a statistical significance (P < 0.05) associated to BoHV-1 infection. These results indicate that BoHV-1 infection is high in the region studied and that factors related to characteristics of the ranches and to the livestock management practices were associated with the risk of infection by BoHV-1. (AU)

A recombinant bovine herpesvirus 5 defective in thymidine kinase and glycoprotein E is immunogenic for calves and confers protection upon homologous challenge and BoHV-1 challenge.

A recombinant bovine herpesvirus 5 lacking thymidine kinase and glycoprotein E genes (BoHV-5gEΔTKΔ) was evaluated as a live experimental vaccine. In a first experiment, ten-months-old calves were vaccinated intramuscularly (n=9) or remained as controls (n=8) and 42 days later were challenged with BoHV-5 or BoHV-1 intranasally. The four control calves challenged with BoHV-5 developed severe depression and neurological signs and were euthanized in extremis at days 13 and 14 pos-infection (pi); the five vaccinated animals challenged with BoHV-5 remained healthy. The titers of virus shedding were reduced (p<0.01) from days 3 to 7 post-infection (pi) in vaccinated animals. Control and vaccinated calves challenged with BoHV-1 presented mild transient respiratory signs; yet the magnitude of virus shedding was reduced (p<0.05) in vaccinated animals (days 5, 9 and 11pi). In a second experiment, young calves (100-120 days-old) were vaccinated (n=15) or kept as controls (n=5) and subsequently challenged with a BoHV-1 isolate. Control calves developed moderate to severe rhinitis and respiratory distress; two were euthanized in extremis at days 5 and 9 pi, respectively. In contrast, vaccinated animals were protected from challenge and only a few developed mild and transient nasal signs. The duration and titers of virus shedding after challenge were reduced (p<0.05) in vaccinated animals comparing to controls. In both experiments, vaccinated animals developed antibodies to gE only after challenge. These results demonstrate homologous and heterologous protection and are promising towards the use of the recombinant BoHV-5gEΔTKΔ in vaccine formulations to control BoHV-5 and BoHV-1 infections.

In vitro-generated interspecific recombinants between bovine herpesviruses 1 and 5 show attenuated replication characteristics and establish latency in the natural host.

BACKGROUND: Interspecific recombinant viruses R1ΔgC and R2ΔgI were isolated after in vitro co-infection with BoHV-1 and BoHV-5, two closely related alphaherpesviruses that infect cattle. The genetic characterization of R1ΔgC and R2ΔgI showed that they are composed of different sections of the parental genomes. The aim of this study was the characterization of the in vivo behavior of these recombinants in the natural host. RESULTS: Four groups of four 3-month-old calves of both genders were intranasally inoculated with either the recombinant or parental viruses. A control group of two animals was also included. Viral excretion and clinical signs were monitored after infection. Histopathological examination of the central nervous system (CNS) was performed and the establishment of latency in trigeminal ganglia was analyzed by PCR. The humoral response was also evaluated using ELISA tests. Three out of four animals from the BoHV-5 infected group excreted virus for 4-10 days. Two calves shed R1ΔgC virus for one day. In R2ΔgI and BoHV-1.2ΔgCΔgI groups, infectious virus was isolated only after two or three blind passages. None of the infected animals developed neurological signs, although those infected with BoHV-5 showed histopathological evidence of viral infection. Latent viral DNA was detected in at least one calf from each infected group. Serum and/or mucosal antibodies were detected in all groups. CONCLUSION: Both BoHV-1/-5 recombinants and the BoHV-1 parental strain are attenuated in calves, although they are able to replicate in animals at low rates and to establish latent infections.

Blood or serum collected on filter paper for detection of antibodies to bovine herpesvirus type 1 (BoHV-1)

The method of collection as well as the packaging conditions in which samples are submitted to laboratories play a critical role on the acquisition of reliable results on diagnostic tests. Alternative methods however have been proposed, as the adsorption of blood or serum in filter paper. In this work, it was evaluated the viability of using serum or whole blood samples from bovines collected in filter paper for serological testing against bovine herpesvirus type 1 (BoHV-1).[...](AU)

Blood or serum collected on filter paper for detection of antibodies to bovine herpesvirus type 1 (BoHV-1)

The method of collection as well as the packaging conditions in which samples are submitted to laboratories play a critical role on the acquisition of reliable results on diagnostic tests. Alternative methods however have been proposed, as the adsorption of blood or serum in filter paper. In this work, it was evaluated the viability of using serum or whole blood samples from bovines collected in filter paper for serological testing against bovine herpesvirus type 1 (BoHV-1).[...]

Development and statistical validation of a guinea pig model for vaccine potency testing against Infectious Bovine Rhinothracheitis (IBR) virus.

Infectious Bovine Rhinothracheitis (IBR) caused by bovine herpesvirus 1 (BoHV-1) infection is distributed worldwide. BoHV-1 either alone or in association with other respiratory cattle pathogens causes significant economic losses to the livestock industry. The aim of this work was to validate a guinea pig model as an alternative method to the current BoHV-1 vaccine potency testing in calves. Guinea pigs were immunized with two doses of vaccine, 21 days apart and sampled at 30 days post vaccination (dpv). BoHV-1 antibody (Ab) response to vaccination in guinea pigs, measured by ELISA and virus neutralization (VN), was statistically compared to the Ab response in cattle. The guinea pig model showed a dose-response relationship to the BoVH-1 antigen concentration in the vaccine and it was able to discriminate among vaccines containing 1log(10) difference in its BoHV-1 concentration with very good repeatability and reproducibility (CV < or = 20%). A regression analysis of the Ab titers obtained in guinea pigs and bovines at 30 and 60dpv, respectively, allowed us to classify vaccines in three potency categories: "very satisfactory", "satisfactory" and "unsatisfactory". Bovines immunized with vaccines corresponding to each of these three categories were experimentally challenged with BoVH-1 virus, the level of protection, as measured by reduction of virus shedding and disease severity, correlated well with the vaccine category used. Data generated by 85 experiments, which included vaccination of calves and guinea pigs with 18 reference vaccines of known potency, 8 placebos and 18 commercial vaccines, was subjected to statistical analysis. Concordance analysis indicated almost perfect agreement between the model and the target species for Ab titers measured by ELISA and almost perfect to substantial agreement when Ab titers were measured by VN. Taken together these results indicate that the developed guinea pig model represents a novel and reliable tool to estimate batch-to-batch vaccine potency and to predict efficacy of killed BoHV-1 veterinary vaccines.