RESUMO
BACKGROUND: Infectious bovine rhinotracheitis (IBR), caused by Bovine alphaherpesvirus-1 (BoAHV-1), is an acute, highly contagious disease primarily characterized by respiratory tract lesions in infected cattle. Due to its severe pathological damage and extensive transmission, it results in significant economic losses in the cattle industry. Accurate detection of BoAHV-1 is of paramount importance. In this study, we developed a real-time fluorescent quantitative PCR detection method for detecting BoAHV-1 infections. Utilizing this method, we tested clinical samples and successfully identified and isolated a strain of BoAHV-1.1 from positive samples. Subsequently, we conducted a genetic evolution analysis on the isolate strain's gC, TK, gG, gD, and gE genes. RESULTS: The study developed a real-time quantitative PCR detection method using SYBR Green II, achieving a detection limit of 7.8 × 101 DNA copies/µL. Specificity and repeatability analyses demonstrated no cross-reactivity with other related pathogens, highlighting excellent repeatability. Using this method, 15 out of 86 clinical nasal swab samples from cattle were found to be positive (17.44%), which was higher than the results obtained from conventional PCR detection (13.95%, 12/86). The homology analysis and phylogenetic tree analysis of the gC, TK, gG, gD, and gE genes of the isolated strain indicate that the JL5 strain shares high homology with the BoAHV-1.1 reference strains. Amino acid sequence analysis revealed that gC, gE, and gG each had two amino acid mutations, while the TK gene had one synonymous mutation and one H to Y mutation, with no amino acid mutations observed in the gD gene. Phylogenetic tree analysis indicated that the JL5 strain belongs to the BoAHV-1.1 genotype and is closely related to American strains such as C33, C14, and C28. CONCLUSIONS: The established real-time fluorescent quantitative PCR detection method exhibits good repeatability, specificity, and sensitivity. Furthermore, genetic evolution analysis of the isolated BoAHV-1 JL-5 strain indicates that it belongs to the BoAHV-1.1 subtype. These findings provide a foundation and data for the detection, prevention, and control Infectious Bovine Rhinotracheitis.
Assuntos
Alphaherpesvirinae , Rinotraqueíte Infecciosa Bovina , Reação em Cadeia da Polimerase em Tempo Real , Rinotraqueíte Infecciosa Bovina/virologia , Animais , Bovinos , Alphaherpesvirinae/classificação , Alphaherpesvirinae/genética , Alphaherpesvirinae/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase em Tempo Real/normas , Sensibilidade e Especificidade , Manejo de Espécimes/veterinária , FilogeniaRESUMO
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.
Assuntos
Anticorpos Neutralizantes/biossíntese , Herpesvirus Bovino 1/imunologia , Vacinas contra Herpesvirus/administração & dosagem , Imunoglobulina G/biossíntese , Rinotraqueíte Infecciosa Bovina/prevenção & controle , Receptor 8 Toll-Like/imunologia , Receptor Toll-Like 9/imunologia , Imunidade Adaptativa/efeitos dos fármacos , Animais , Anticorpos Antivirais , Bovinos , Proliferação de Células , Endossomos/imunologia , Endossomos/metabolismo , Expressão Gênica , Herpesvirus Bovino 1/patogenicidade , Imunidade Inata/efeitos dos fármacos , Imunização Secundária/métodos , Rinotraqueíte Infecciosa Bovina/genética , Rinotraqueíte Infecciosa Bovina/imunologia , Rinotraqueíte Infecciosa Bovina/virologia , Interferon gama/genética , Interferon gama/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Linfócitos/imunologia , Linfócitos/virologia , Masculino , Cavidade Nasal/imunologia , Cavidade Nasal/virologia , Receptor 8 Toll-Like/agonistas , Receptor 8 Toll-Like/genética , Receptor Toll-Like 9/agonistas , Receptor Toll-Like 9/genética , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Vacinação/métodos , Vacinas de Produtos InativadosRESUMO
Herpesviruses are the most prevalent viruses that infect the human and animal body. They can escape a host immune response in numerous ways. One way is to block the TAP complex so that viral peptides, originating from proteasomal degradation, cannot be transported to the endoplasmic reticulum. As a result, a reduced number of MHC class I molecules appear on the surface of infected cells and, thus, the immune system is not efficiently activated. BoHV-1-encoded UL49.5 protein is one such TAP transporter inhibitor. This protein binds to TAP in such a way that its N-terminal fragment interacts with the loops of the TAP complex, and the C-terminus stimulates proteasomal degradation of TAP. Previous studies have indicated certain amino acid residues, especially the RRE(9-11) motif, within the helical structure of the UL49.5 N-terminal fragment, as being crucial to the protein's activity. In this work, we investigated the effects of modifications within the RRE region on the spatial structure of the UL49.5 N-terminal fragment. The introduced RRE(9-11) variations were designed to abolish or stabilize the structure of the α-helix and, consequently, to increase or decrease protein activity compared to the wild type. The terminal structure of the peptides was established using circular dichroism (CD), 2D nuclear magnetic resonance (NMR), and molecular dynamics (MD) in membrane-mimetic or membrane-model environments. Our structural results show that in the RRE(9-11)AAA and E11G peptides the helical structure has been stabilized, whereas for the RRE(9-11)GGG peptide, as expected, the helix structure has partially unfolded compared to the native structure. These RRE modifications, in the context of the entire UL49.5 proteins, slightly altered their biological activity in human cells.
Assuntos
Infecções por Herpesviridae/virologia , Herpesvirus Bovino 1/química , Rinotraqueíte Infecciosa Bovina/virologia , Proteínas do Envelope Viral/química , Motivos de Aminoácidos , Animais , Bovinos , Humanos , Modelos Moleculares , Fragmentos de Peptídeos/química , Conformação Proteica , Estabilidade ProteicaRESUMO
Many studies report age as a risk factor for BoHV-1 infection or seropositivity. However, it is unclear whether this pattern reflects true epidemiological causation or is a consequence of study design and other issues. Here, we seek to understand the age-related dynamics of BoHV-1 seroprevalence in seasonal calving Irish dairy herds and provide decision support for the design and implementation of effective BoHV-1 testing strategies. We analysed seroprevalence data from dairy herds taken during two Irish seroprevalence surveys conducted between 2010 and 2017. Age-dependent seroprevalence profiles were constructed for herds that were seropositive and unvaccinated. Some of these profiles revealed a sudden increase in seroprevalence between adjacent age-cohorts, from absent or low to close to 100% of seropositive animals. By coupling the outcome of our data analysis with simulation output of an individual-based model at the herd scale, we have shown that these sudden increases are related to extensive virus circulation within a herd for a limited time, which may then subsequently remain latent over the following years. BoHV-1 outbreaks in dairy cattle herds affect animals independent of age and lead to almost 100% seroconversion in all age groups, or at least in all animals within a single epidemiological unit. In the absence of circulating infection, there is a year-on-year increase in the age-cohort at which seroprevalence changes from low to high. The findings of this study inform recommendations regarding testing regimes in the context of contingency planning or an eradication programme in seasonal calving dairy herds.
Assuntos
Herpesvirus Bovino 1/fisiologia , Rinotraqueíte Infecciosa Bovina/epidemiologia , Vacinação/veterinária , Fatores Etários , Animais , Bovinos , Indústria de Laticínios , Feminino , Rinotraqueíte Infecciosa Bovina/virologia , Irlanda/epidemiologia , Prevalência , Estudos SoroepidemiológicosRESUMO
This study aimed to determine the seroprevalence and identify the risk factors associated with Neospora caninum, Bovine herpesvirus type 1 (BHV-1), and Bovine viral diarrhea virus (BVDV) infection on industrial Holstein dairy cattle farms in Isfahan province, Central Iran. Blood samples were taken from 216 apparently healthy cattle from 16 randomly selected Holstein dairy farms in the North, South, East, and West of Isfahan in the summer of 2017. The antibodies to N. caninum, BHV-1, and BVDV were detected using a commercially available ELISA kit. The overall seroprevalence for N. caninum, BHV-1, and BVDV was 19%, 72.2%, and 52.8%, respectively. The significant major risk factors of BHV-1 in cattle were identified as farm direction, age groups, parity, and milk yield by the univariate analysis (p < 0.05). The significant major risk factors of BVDV in cattle were identified as age groups, parity, milk yield, and stage of pregnancy (p < 0.05). The only significant major risk factor of N. caninum was farm direction (p < 0.05). A significant association of concurrent infection with BVDV and BHV-1 has shown in the current study (p < 0.05). This study is the first to report the risk factors for N. caninum, BHV-1, and BVDV infection in the central part of Iran and allows us to conclude that these agents are widely distributed in this region.
Assuntos
Aborto Animal/epidemiologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Doenças dos Bovinos/epidemiologia , Coccidiose/veterinária , Infecções por Herpesviridae/veterinária , Aborto Animal/parasitologia , Aborto Animal/virologia , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Doenças dos Bovinos/virologia , Coccidiose/epidemiologia , Coccidiose/parasitologia , Vírus da Diarreia Viral Bovina/fisiologia , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Herpesvirus Bovino 1/fisiologia , Rinotraqueíte Infecciosa Bovina/epidemiologia , Rinotraqueíte Infecciosa Bovina/virologia , Irã (Geográfico)/epidemiologia , Neospora/fisiologia , Prevalência , Fatores de Risco , Estudos SoroepidemiológicosRESUMO
Camels (Camelus dromedarius) are bred in Western Turkey, particularly in the province of Aydin, for touristic, social and cultural purposes. Bovine enterovirus1 (BEV1), Bovine herpesvirus type1 (BHV1), Bovine viral diarrhea virus (BVDV), and Parainfluenza3 (PI3) virus infections are significant causes of health and/or economic concerns in several animal species. These agents have not been investigated in the camel population in Turkey. The objective of this study was to serologically investigate the presence and infection rates of these viruses in camels in Aydin province, Western Turkey. Ninetytwo serum samples were taken from clinically healthy camels that were kept in private farms or brought to the local slaughterhouses. Serum neutralization test was performed to assess the presence and the titers of specific antibodies against BEV1, BHV1, BVDV, and PI3 virus in camel sera. Of the 92 camels tested, 30 (32.61%), 2 (2.17%), 54 (58.7%), and 20 (21.74%) were seropositive for BEV1, BHV1, BVDV, and PI3, respectively. These results suggest that, except for BHV1, these viral infections are common among camels in Western Turkey. To our knowledge, this the first comprehensive, largescale study investigating these viral infections in camels in Turkey.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Camelus , Infecções por Enterovirus/epidemiologia , Rinotraqueíte Infecciosa Bovina/epidemiologia , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Vírus da Diarreia Viral Bovina/isolamento & purificação , Infecções por Enterovirus/sangue , Infecções por Enterovirus/virologia , Enterovirus Bovino/isolamento & purificação , Feminino , Herpesvirus Bovino 1/isolamento & purificação , Rinotraqueíte Infecciosa Bovina/sangue , Rinotraqueíte Infecciosa Bovina/virologia , Masculino , Turquia/epidemiologiaRESUMO
The aim of the study was to determine the antibody sero-prevalence of Bovine Herpesvirus-1 which cause Infectious Bovine Rhinotracheitis (IBR) and to identify risk factors associated with BHV-1 antibody seropositivity among smallholder dairy farms in Meru County, Kenya. A cross-sectional study was conducted in the Naari area of Meru County, Kenya between September-October 2016 and March-April 2017. The 149 farmers were randomly selected from members of the Naari Dairy Farmers Cooperative Society who were actively delivering milk to the society at the time of the study. Serum samples were obtained from 403 female dairy cattle. Farm level management and animal factors were collected through direct interviews with the owner or someone who was knowledgeable about the animals. All serum samples were processed with an indirect enzyme-linked immunosorbent assay (gB ELISA) to determine the presence of antibodies to BHV-1. The overall farm-level and animal-level sero-prevalences of BHV-1 antibodies were 30.9 % (95 % CI: 23.6%-39.0%) and 17.4 % (95 % CI: 13.8%-21.4%), respectively. In the final multivariable analysis, the factors significantly associated with BHV-1 antibodies included; age of the dairy cattle (ORâ¯=â¯1.200, pâ¯=â¯0.001), age of the principal female farmers (ORâ¯=â¯0.182, pâ¯=â¯0.001) and rearing goats in the farm (ORâ¯=â¯26.77, pâ¯=â¯0.000). There was a significant interaction between rearing goats on the farm and age of the dairy cattle (pâ¯<â¯0.010); younger cattle seemed to have been exposed to BHV or a cross-reacting caprine herpesvirus when goats were on the farm. The results showed that BHV-1 was circulating among the cattle population in the Naari area of Meru County. Given that there is not BHV-1 vaccination use in this study population, training on the importance of biosecurity and vaccination for BHV-1 are recommended to reduce the transmission and impacts of BHV-1.
Assuntos
Anticorpos Antivirais/sangue , Herpesvirus Bovino 1/isolamento & purificação , Rinotraqueíte Infecciosa Bovina/epidemiologia , Animais , Bovinos , Estudos Transversais , Indústria de Laticínios , Feminino , Rinotraqueíte Infecciosa Bovina/virologia , Quênia/epidemiologia , Prevalência , Fatores de Risco , Estudos SoroepidemiológicosRESUMO
Bovine alphaherpesvirus 1 (BoHV-1) as a cusative agent for some diseases in cattle infects sheep and goat; and it is believed that these animals may be reservoir host for this virus. Thus, BoHV-1 infection in sheep and goat should be considerd when there is a program for control and eradication of this virus in cattle. Therefore, the aim of this study was to determine the seroprevalence of BoHV-1 in sheep, relationship between host and environmental factors with infection, and the role of sheep in the epidemiology of the BoHV-1. Blood samples were randomly collected from 310 healthy sheep in 6 cities of Khuzestan province (Southwest of Iran) including Ahvaz, Hendijan, Shushtar, Dezful, Masjed Soleyman and Behbahan. Sera were analyzed by virus neutralisation (VN) test for detection antibodies to BoHV-1. According to VN test, apparent and true seroprevalence seroprevalence of BoHV-1 infection was 28.4 % (95%CI: 23.4-33.4%) and 28.4 % (95%CI: 23.3-33.4%), respectively. Logistic regression revealed that the odds of infection between the age was 1.06 (95%CI: 0.9-1.25) (P > 0.05), implying that the odds of infection increased 6 % with rising one year of age. Besides, the relative frequency of infection in males was more than females', and the odds of infection in male sheep was identified to be 1.13 (95%CI: 0.47-2.71) (P > 0.05), compared to that in females. Moreover, in comparison to sheep with no history of abortion, the odds of infection in sheep with a history of abortion was 1.28 (95%CI: 0.57-2.87) (P > 0.05). The seroprevalence in Shushtar, Masjed Soleyman, Dezful, Ahvaz, Hendijan, and Behbahan were found to be 48.3, 46.7, 31.7, 20, 16.7, and 12 percent, respectively and 13.1 of fluctuation in infection can be justified by different geographical locations investigated in this study (P < 0.001). Considering the significant seroprevalence of BoHV-1, present study confirmed the role of sheep in the epidemiology of this virus and control of BoHV-1 in sheep should be considered by animal health authorities in areas where BoHV-1 is prevalent.
Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 1/fisiologia , Rinotraqueíte Infecciosa Bovina/epidemiologia , Doenças dos Ovinos/epidemiologia , Animais , Bovinos , Feminino , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Rinotraqueíte Infecciosa Bovina/virologia , Irã (Geográfico)/epidemiologia , Masculino , Prevalência , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/virologiaRESUMO
Bovine herpesvirus type 1 (BoHV-1), a member of the Alphaherpesvirinae subfamily, causes significant economic losses to the cattle industry worldwide. Envelope glycoprotein D (gD) of BoHV-1 plays an essential role in the viral entry into permissive cells and possibly cooperates with other envelope glycoproteins. The herpesvirus gD induces a protective immune response against diseases in cattle or animal models. Mapping epitopes on gD will facilitate the understanding of the BoHV-1 pathogenesis and development of alternative vaccines against various diseases associated with the virus. In this study, a monoclonal antibody (MAb), designated as 3C1, was generated using naive BoHV-1 in vaccination of mice, demonstrating that 3C1 was specific to gD and represents a neutralizing activity against BoHV-1 infection in Madin-Darby bovine kidney cells. Panels of overlapping gD recombinant proteins with glutathione S-transferase tag were prepared to define the epitope recognized by 3C1. The data demonstrated that the N-terminus of gD 23APRVTVYVD31 was recognized by 3C1. Furthermore, the 26VTVYVD31 motif was the minimal amino acid sequence for the recognition. The epitope identified in this study is highly conserved among the typical strains of BoHV-1 and BoHV-5, suggesting that this epitope may be useful in the diagnosis of diseases. In addition, the defined region on gD of BoHV-1 might be essential in viral entry upon comparison with the prototype virus in herpes simplex virus (Alphaherpesvirinae). The data will elucidate the roles of gD of BoHV-1 in viral entry and pathogenesis and its potential application for the development of vaccine candidates and diagnostic techniques based on the conserved epitopes on gD or in combination with those of other herpesvirus glycoproteins.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos Virais/imunologia , Mapeamento de Epitopos , Epitopos/imunologia , Herpesvirus Bovino 1/imunologia , Rinotraqueíte Infecciosa Bovina/virologia , Proteínas Virais/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/imunologia , Antígenos Virais/química , Bovinos , Linhagem Celular , Epitopos/química , Masculino , Camundongos , Testes de Neutralização , Proteínas Recombinantes , Proteínas Virais/químicaRESUMO
Bovine respiratory disease (BRD) remains a major health problem despite extensive use of vaccines during the post-weaning period. Apparent vaccine failure is attributed, in part, to primary vaccination during the period of greatest risk for BRD, providing inadequate time for onset of protective immunity. The current study investigated whether intranasal (IN) vaccination of 3-6â¯week old calves with a modified-live viral (MLV) vaccine induced sufficient immune memory to prevent respiratory disease and accelerate onset of protective immunity 5â¯months later. Vaccine groups included naïve controls, a single IN vaccination at 3-6â¯weeks of age, primary IN vaccination at 6â¯months, and either an IN or subcutaneous (SC) booster vaccination at 6â¯months (nâ¯=â¯10/group). All calves were challenged with BHV-1 four days after vaccination at 6â¯months of age. Primary IN vaccination at 6â¯months did not significantly reduce clinical disease but significantly (Pâ¯<â¯0.01) reduced virus shedding. A single IN vaccination at 3-6â¯weeks of age significantly (Pâ¯<â¯0.05) reduced weight loss but did not reduce fever or virus shedding. Both IN and SC booster vaccinations, significantly (Pâ¯<â¯0.01) reduced clinical disease but virus shedding was significantly (Pâ¯<â¯0.001) reduced only by IN booster vaccination. Reduction in virus shedding was significantly (Pâ¯<â¯0.01) greater following booster versus primary IN vaccination at 6â¯months. All vaccination regimes significantly (Pâ¯<â¯0.01) reduced secondary bacterial pneumonia and altered interferon responses relative to naïve controls. Only IN booster vaccination significantly (Pâ¯<â¯0.05) increased BHV-1 specific IgA in nasal secretions. These results confirm primary MLV IN vaccination at 3 to 6â¯weeks of age, when virus neutralizing maternal antibody was present, induced immune memory with a 5â¯month duration. This immune memory supported rapid onset of protective immunity four days after an IN booster vaccination.
Assuntos
Herpesvirus Bovino 1/imunologia , Vacinas contra Herpesvirus/administração & dosagem , Imunização Secundária/métodos , Memória Imunológica/efeitos dos fármacos , Rinotraqueíte Infecciosa Bovina/prevenção & controle , Pneumonia Bacteriana/prevenção & controle , Administração Intranasal , Animais , Animais Recém-Nascidos , Anticorpos Antivirais/sangue , Bovinos , Colostro/química , Colostro/imunologia , Feminino , Herpesvirus Bovino 1/efeitos dos fármacos , Herpesvirus Bovino 1/patogenicidade , Imunidade nas Mucosas/efeitos dos fármacos , Imunoglobulina A/sangue , Rinotraqueíte Infecciosa Bovina/imunologia , Rinotraqueíte Infecciosa Bovina/mortalidade , Rinotraqueíte Infecciosa Bovina/virologia , Masculino , Pneumonia Bacteriana/imunologia , Pneumonia Bacteriana/microbiologia , Pneumonia Bacteriana/mortalidade , Gravidez , Análise de Sobrevida , Vacinação/métodos , Vacinas Atenuadas , Carga Viral/efeitos dos fármacos , Eliminação de Partículas Virais/efeitos dos fármacos , Redução de Peso/efeitos dos fármacosRESUMO
The Cooper and Los Angeles (LA) strains were the two original respiratory strains of bovine herpesvirus type 1.1 (BoHV-1.1) isolated in the 1950s from cattle with infectious bovine rhinotracheitis. We report the complete genome sequence for the BoHV-1.1 LA strain and compare it to the prototype Cooper strain and six wild-type BoHV-1.1 isolates. A nucleotide sequence divergence of 0.74% was noted across the two complete genomes, caused by 19 single-nucleotide polymorphisms (SNPs) involving 12 genes and insertions/deletions that primarily affected the number of repeats within reiterated repeat regions of the genome. Phylogenetic analysis revealed that Cooper and LA strains are genetically the most ancient strains from which all of the more-recently isolated field strains of BoHV-1.1 evolved.
Assuntos
Genoma Viral/genética , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 1/genética , Rinotraqueíte Infecciosa Bovina/virologia , Animais , Sequência de Bases , Bovinos , Doenças dos Bovinos/virologia , Genótipo , Herpesvirus Bovino 1/classificação , Polimorfismo de Nucleotídeo Único/genética , Análise de Sequência de DNA , Sequenciamento Completo do GenomaRESUMO
Bovine herpesvirus type 1 and type 5 (BoHV-1 and BoHV-5) are two alphaherpesviruses that affect cattle with two different syndromes. While BoHV-1 mainly produces respiratory symptoms, BoHV-5 is highly neuropathogenic and responsible for meningoencephalitis in young cattle. The latency-related (LR) gene, which is not conserved between these two herpesviruses, is the only viral gene abundantly expressed in latently infected neurons. The antiapoptotic action of this gene has been demonstrated during acute infection and reactivation from latency and seems to be mainly mediated by a LR protein (ORF-2) which is truncated in amino acid 51 in the case of BoHV-5. In this work, we show that the BoHV-5 LR gene is less efficient at cell survival and apoptosis inhibition in transient as well as in established neuronal cell lines compared to its BoHV-1 homolog. We hypothesize that the BoHV-5 LR gene may have novel functions that are lacking in the BoHV-1 LR gene and that these differences may contribute to its enhanced neuropathogenesis.
Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 5/genética , Rinotraqueíte Infecciosa Bovina/metabolismo , Meningoencefalite/veterinária , Proteínas Virais/genética , Latência Viral/genética , Animais , Apoptose/genética , Bovinos , Linhagem Celular , Expressão Gênica , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologia , Herpesvirus Bovino 1/crescimento & desenvolvimento , Herpesvirus Bovino 1/metabolismo , Herpesvirus Bovino 5/crescimento & desenvolvimento , Herpesvirus Bovino 5/metabolismo , Interações Hospedeiro-Patógeno/genética , Rinotraqueíte Infecciosa Bovina/patologia , Rinotraqueíte Infecciosa Bovina/virologia , Meningoencefalite/patologia , Meningoencefalite/virologia , Neurônios/metabolismo , Neurônios/virologia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteínas Virais/metabolismo , Ativação ViralRESUMO
Bovine herpesvirus 1 (BoHV1) is the cause of economically significant viral infections in cattle. Respiratory symptoms associated with the infection are known as Infectious Bovine Rhinotracheitis (IBR). Sheep and goats are less sensitive to the infection although their role in inter-species viral transmission under field conditions is subject to controversy. The objective of this study was to investigate seroprevalence of BoHV1 infections in cattle, sheep, and goats raised together for at least a year. Blood serum samples were taken from 226 cattle, 1.053 sheep, and 277 goats from 17 small- to medium-scale farms. BoHV1-specific antibody presence and titers were determined using virus neutralization test. In total, 73 of the 226 cattle (32.3%) were seropositive. The infection was detected in 13 of the 17 farms. Infection rates ranged from 5.8 to 88.8%. Only one of the 1053 sheep (0.09%) was seropositive. However, 58 of the 277 (20.9%) goats were seropositive. Goat samples taken from 8 of the 17 farms were seropositive with infection rates ranging from 17 to 38.9%. Statistical analysis showed a significant correlation in infection rates between cattle and goats but not sheep. These results suggest that goats may be more sensitive to the BHV1 infection than sheep and the role of goats as possible reservoirs for BoHV1 in the control and eradication of BHV1 in cattle should be considered in future studies.
Assuntos
Herpesvirus Bovino 1/imunologia , Rinotraqueíte Infecciosa Bovina/epidemiologia , Animais , Anticorpos Antivirais/sangue , Bovinos , Reservatórios de Doenças/veterinária , Doenças das Cabras/epidemiologia , Doenças das Cabras/transmissão , Doenças das Cabras/virologia , Cabras , Rinotraqueíte Infecciosa Bovina/transmissão , Rinotraqueíte Infecciosa Bovina/virologia , Testes de Neutralização/veterinária , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/transmissão , Doenças dos Ovinos/virologia , Turquia/epidemiologiaRESUMO
Following acute infection of mucosal surfaces by bovine herpesvirus 1 (BoHV-1), sensory neurons are a primary site for lifelong latency. Stress, as mimicked by the synthetic corticosteroid dexamethasone, consistently induces reactivation from latency. Two viral regulatory proteins (VP16 and bICP0) are expressed within 1 h after calves latently infected with BoHV-1 are treated with dexamethasone. Since the immediate early transcription unit 1 (IEtu1) promoter regulates both BoHV-1 infected cell protein 0 (bICP0) and bICP4 expressions, we hypothesized that the bICP4 protein is also expressed during early stages of reactivation from latency. In this study, we tested whether bICP4 and bICP22, the only other BoHV-1 protein known to be encoded by an immediate early gene, were expressed during reactivation from latency by generating peptide-specific antiserum to each protein. bICP4 and bICP22 protein expression were detected in trigeminal ganglionic (TG) neurons during early phases of dexamethasone-induced reactivation from latency, operationally defined as the escape from latency. Conversely, bICP4 and bICP22 were not readily detected in TG neurons of latently infected calves. In summary, it seems clear that all proteins encoded by known BoHV-1 IE genes (bICP4, bICP22, and bICP0) were expressed during early stages of dexamethasone-induced reactivation from latency.
Assuntos
Regulação Viral da Expressão Gênica , Herpesvirus Bovino 1/genética , Proteínas Imediatamente Precoces/genética , Rinotraqueíte Infecciosa Bovina/virologia , Células Receptoras Sensoriais/virologia , Gânglio Trigeminal/virologia , Proteínas Virais/genética , Animais , Anticorpos Antivirais/química , Bovinos , Linhagem Celular , Dexametasona/farmacologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Células Epiteliais/virologia , Herpesvirus Bovino 1/crescimento & desenvolvimento , Herpesvirus Bovino 1/metabolismo , Proteínas Imediatamente Precoces/metabolismo , Rinotraqueíte Infecciosa Bovina/patologia , Rim/efeitos dos fármacos , Rim/patologia , Rim/virologia , Masculino , Células Receptoras Sensoriais/efeitos dos fármacos , Células Receptoras Sensoriais/metabolismo , Células Receptoras Sensoriais/patologia , Gânglio Trigeminal/efeitos dos fármacos , Gânglio Trigeminal/metabolismo , Gânglio Trigeminal/patologia , Proteínas Virais/metabolismo , Ativação Viral/efeitos dos fármacos , Latência Viral/efeitos dos fármacosRESUMO
Bovine herpesvirus 1 (BoHV-1) is one of several microbes that contributes to the development of the bovine respiratory disease (BRD) and can also induce abortions in cattle. As other alpha-herpesvirinae subfamily members, BoHV-1 efficiently replicates in many cell types and subsequently establishes a life-long latent infection in sensory neurons. BoHV-1 encodes more than 70 proteins that are expressed in a well-defined manner during productive infection. However, in silico open reading frame (ORF) prediction of the BoHV-1 genome suggests that the virus may encode more than one hundred proteins. In this study we used mass spectrometry followed by proteogenomic mapping to reveal the existence of 92 peptides that map to previously un-annotated regions of the viral genome. Twenty-one of the newly termed "intergenic peptides" were predicted to have a viable ORF around them. Twelve of these produced an mRNA transcript as demonstrated by strand-specific RT-PCR. We further characterized the 5' and 3' termini of one mRNA transcript, ORF-A, and detected a 55 kDa protein produced during active infection using a custom-synthesized antibody. We conclude that the coding potential of BoHV-1 is underestimated.
Assuntos
Regulação Viral da Expressão Gênica , Herpesvirus Bovino 1/fisiologia , Rinotraqueíte Infecciosa Bovina/virologia , Proteogenômica , Proteínas Virais/genética , Proteínas Virais/metabolismo , Animais , Sequência de Bases , Bovinos , Linhagem Celular , Células Cultivadas , Cromatografia Líquida , Fases de Leitura Aberta , Peptídeos/genética , Peptídeos/metabolismo , Proteogenômica/métodos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem , Proteínas Virais/química , Replicação ViralRESUMO
Infectious bovine rhinotracheitis (IBR)/infectious pustular vulvovaginitis (IPV) caused by Bovine alphaherpesvirus 1 (BoHV-1) is a significant disease in domestic and wild cattle. In June 2015, the Ministry of Agriculture, Food and Forestry in Italy approved a national surveillance plan to control and eradicate IBR in beef cattle breeds. The objective of this study was to evaluate the results of the first year of the IBR voluntary surveillance plan in Italy. The aim of the plan is to eradicate IBR in all bovines recorded in the National Herd Book for Italian beef cattle breeds over six years. Monetary incentives are used to encourage breeders to achieve the annual seroprevalence ranges stated in the plan. A Ministerial decree states that all bovines in breeding herds and aged older than 12 months should be serologically tested. Serum samples were tested for presence of the antibody to glycoprotein E of BoHV-1 using commercially available enzyme-linked immunosorbent assays. The national herd seroprevalence was 55.49% (95% confidence interval [CI] 52.01-58.92). Of 25,121 bovines tested for antibodies against BoHV-1, 8014 were positive. The seroprevalence in animals from autochthonous Italian cattle breeds was 31.89% (95% CI 31.31-32.47). Seroprevalence was highest in Podolica cattle (55.14%; 95% CI 54.07-56.21), lowest in Maremmana cattle (9.95%; 95% CI 7.99-12.31), and intermediate in Chianina (22.01%; 95% CI 21.03-23.01), Marchigiana (24.85%; 95% CI 23.52-26.23), and Romagnola (15.60%; 95% CI 14.62-16.64) cattle. These seroprevalence rates indicate a need for intervention to decrease the inevitable severe economic losses arising from BoHV-1 infection. Although some regions in Italy have a long history of combatting BoHV-1 infection, only the province of Bolzano has eradicated IBR.
Assuntos
Anticorpos Antivirais/sangue , Erradicação de Doenças/estatística & dados numéricos , Monitoramento Epidemiológico/veterinária , Rinotraqueíte Infecciosa Bovina/epidemiologia , Rinotraqueíte Infecciosa Bovina/imunologia , Alphaherpesvirinae/imunologia , Alphaherpesvirinae/isolamento & purificação , Animais , Animais Domésticos , Cruzamento , Bovinos , Erradicação de Doenças/métodos , Ensaio de Imunoadsorção Enzimática , Feminino , Rinotraqueíte Infecciosa Bovina/virologia , Itália/epidemiologia , Estudos SoroepidemiológicosRESUMO
Bovine herpesvirus type 1 (BoHV-1) is recognized as an important pathogen causing respiratory, reproductive, and neurological disorders in cattle and is associated with economic losses to animal industry. Accurate diagnostic methods are needed for prevention of disease transmission. While the virus neutralization test is considered the gold standard method, it requires maintenance of the virus and cell cultures, which is time consuming and expensive. Serological techniques such as enzyme-linked immunosorbent assay (ELISA) are widely applied, as these are easy to perform and provide quick results. In the present study, a nanogold slot blot inhibition assay was developed for the serological diagnosis of BoHV-1 and compared with standard ELISA and horseradish peroxidase (HRP) slot blot assays. Of 42 serum samples tested by ELISA, 32 (76.2%) were positive and 10 (23.8%), were negative. The sensitivity and specificity of the nanogold slot blot inhibition assay was similar to that observed for ELISA and HRP slot blot assays, and a strong correlation was observed between the tests. Thus, the nanogold slot blot inhibition assay may serve as an efficient and rapid alternative to ELISA in settings, where plate-reading equipment is lacking.
Assuntos
Anticorpos Antivirais/química , Bioensaio , Western Blotting/métodos , Coloide de Ouro/química , Herpesvirus Bovino 1/imunologia , Rinotraqueíte Infecciosa Bovina/diagnóstico , Animais , Anticorpos Antivirais/isolamento & purificação , Benchmarking , Western Blotting/instrumentação , Bovinos , Cães , Ensaio de Imunoadsorção Enzimática , Herpesvirus Bovino 1/isolamento & purificação , Soros Imunes/química , Imunoconjugados/química , Rinotraqueíte Infecciosa Bovina/sangue , Rinotraqueíte Infecciosa Bovina/virologia , Células Madin Darby de Rim Canino , Nanopartículas Metálicas/química , Sensibilidade e EspecificidadeRESUMO
Bovine herpesvirus-1 (BHV-1) causes infectious bovine rhinotracheitis (IBR), and infectious pustular vulvovaginitis (IPV) in cows and infectious pustular balanopostitis (IPB) in bulls worldwide. Infection of seronegative cattle with BHV-1 leads to abortion, retention of fetal membranes, increased service per conception, metritis and oophoritis. As part of an ongoing study on infectious causes of reproductive disorders in Ethiopia, this investigation aims at assessing the role of BHV-1 in the disorders and the risk factors affecting its seroprevalence. A cross-sectional study was conducted on a total of 1379 randomly selected dairy cattle from 149 herds. These dairy cattle were sampled from milks sheds of central (nâ¯=â¯555), western (nâ¯=â¯195) and southern (nâ¯=â¯629) Ethiopia. Blocking enzyme-linked immunosorbent assay (B-ELISA) was applied to detect antibodies specific to BHV-1. Additionally, a semi-structured questionnaire was administered and farm records were assessed to capture potential risk factors associated with BHV-1 seropositivity. Univariable and multivariable random-effects logistic regression analyses were used to assess potential risk factors associated with BHV-1 serostatus. Model fitness and reliability were assessed using the Hosmer and Lemeshow method and the receiver operating curve (ROC) respectively. An overall herd level BHV-1 seroprevalence of 81.8% (95% confidence interval (CI): 74.7-87.7%) and individual animal level seroprevalence of 41.0% (95% CI: 38.4-43.7%) were found. In a random-effects multivariable logistic regression model, the seroprevalence of BHV-1 exposure was higher in dairy cattle from breeding (Odds ratio [OR]â¯=â¯1.3; pâ¯=â¯0.036) than in commercial (ORâ¯=â¯0.9; pâ¯=â¯0.137) and small-holder farms. Geographically, the prevalence was higher in western (ORâ¯=â¯1.4; pâ¯<â¯0.001) and southern Ethiopia (ORâ¯=â¯1.2; pâ¯<â¯0.001) than in central regions. BHV-1 seropositive cows had higher (pâ¯<â¯0.05) odds of clinical reproductive disorders including abortion, retained fetal membranes, stillbirth, birth of weak calf and metritis compared to seronegative cows. Thus, it is suggested that BHV-1 should be considered as differential diagnosis among improved dairy cattle herds with reproductive disorders in Ethiopia.
Assuntos
Herpesvirus Bovino 1/isolamento & purificação , Rinotraqueíte Infecciosa Bovina/epidemiologia , Doenças do Pênis/veterinária , Vulvovaginite/veterinária , Animais , Bovinos , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Etiópia/epidemiologia , Feminino , Rinotraqueíte Infecciosa Bovina/virologia , Masculino , Doenças do Pênis/epidemiologia , Doenças do Pênis/virologia , Prevalência , Fatores de Risco , Estudos Soroepidemiológicos , Vulvovaginite/epidemiologia , Vulvovaginite/virologiaRESUMO
A follow-up study from 2005 to 2010 was carried out in two herds where eradication programme for the bovine herpes virus-1 (BoHV-1) infection depends on the vaccination with inactivated glycoprotein E-deleted vaccine that was started in 2001 following the vaccination with inactivated conventional vaccine between 1999 and 2001. For serological screening, a total of 12,976 sera sampled over several sampling times approximately 6 months of interval during 5 years (2005-2010) were tested for glycoprotein E (gE)- and glycoprotein B-specific antibodies using ELISA. According to the serological evidence, the long-term persistence of BoHV-1 antibodies, success of marker vaccine, first vaccination time of the calves in herds regularly vaccinated, etc. were discussed in this paper. In conclusion, the vaccination programme using gE (-) marker vaccines, with making efforts to prevent the other factors about transmission of infection, was suggested for the eradication of BoHV-1 infection in Turkey as many EU countries. This is the first report on the BoHV-1 eradication programme in some dairy cattle in Turkey.
Assuntos
Herpesvirus Bovino 1/imunologia , Rinotraqueíte Infecciosa Bovina/prevenção & controle , Proteínas Virais/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Bovinos , Doenças dos Bovinos/prevenção & controle , Indústria de Laticínios , Ensaio de Imunoadsorção Enzimática , Feminino , Seguimentos , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/veterinária , Rinotraqueíte Infecciosa Bovina/virologia , Estudos Longitudinais , Projetos Piloto , Turquia , Vacinação , Vacinas de Produtos Inativados , Vacinas Marcadoras/imunologiaRESUMO
In the Netherlands, the feasibility of a national control program for infectious bovine rhinotracheitis (IBR) is discussed. The aim of this program would be to achieve freedom from BoHV1 circulation (the causal agent of IBR), in the Dutch cattle population. When IBR would be eradicated, maintaining the free status is essential and insight in the probability of introduction of IBR through cattle imports is crucial. Values for input parameters such as the number of imports per country of origin, herd level prevalence and probability that a random imported animal per age category was either acutely or latently infected with IBR were quantified. A stochastic simulation model was built to predict the basic risk and the efficacy of four risk mitigating scenarios were evaluated. These scenarios involved testing prior to import, import restrictions and vaccination. The model output predicted that IBR infected animals are imported regularly. In an IBR free situation, 571 (5th and 95th percentile: 431-781) cattle herds will be newly infected. Latent infections account for most newly infected herds (77%). When the virus in the imported latently infected animal does not reactivate, subsequent impact of such infections remains limited. The model predicted that most of the herds infected by introduction of acutely infected animals would be veal herds. The scenario in which imports were only allowed from status 9 or 10 countries combined with testing cattle that originated from status 9 countries was most effective in reduction of the import risk to 70 herds per year. The scenario in which vaccination of calves was combined with testing of older cattle was estimated to reduce the number of newly infected herds to 82 per year. The stakeholders classified the latter scenario as most realistic because this scenario was deemed both feasible and rather effective. This study did not evaluate the impact of introduction of IBR in the cattle population, which might differ depending on the type of infection (acute vs. latent) and the herd type in which the virus is introduced. Moreover, when making the final decision about the optimal intervention, the economic perspective should also be taken into account. This study predicted that introduction of IBR will remain a risk for the Dutch cattle population after virus circulation is eliminated from the Netherlands. The import risk is reduced most in scenarios in which testing and vaccination are combined.