RESUMEN
BACKGROUND: Widely regarded as one of the chief causes of diseases in cattle population, bovine herpesvirus-1 (BoHV-1) has the potential to infect sheep and goat, making them potential reservoirs or hosts for this virus. Thus, preventive measures against BoHV-1 in cattle should not overlook the ability of this virus to infect other animals. AIMS: Therefore, the focal point of this study was to ascertain the seroprevalence of BoHV-1 in 300 healthy goats, the relationship between host and the environmental determinants of infection, and the contributing role of goats in the epidemiology of the BoHV-1. MATERIALS & METHODS: In order to pinpoint the existing antibodies to BoHV-1, the obtained sera were analyzed by Virus Neutralization test. RESULTS: According to this test, the seroprevalence of BoHV-1 appeared to be 64.33% in southwestern Iran. What logistic regression disclosed was that the odds ratio between age and infection with BoHV-1 was 0.83 (p = 0.01), representing a decrease of 17% as goats grew one year older. In addition, females manifested a higher relative frequency of infection compared to males, with the odds of infection in female goats being registered at 1.88, compared to those in males (p = 0.2). Moreover, contrasted with goats lacking any history of abortion, those with a history of abortion featured 1.1 as the odds ratio (p = 0.87). The seroprevalence in Hendijan, Ahvaz, Shushtar and Dasht e Azadegan was detected to stand at 73.24, 71.30, 55.56 and 47.06 percent, respectively, with 6% of fluctuation in the infection rates being attributed to various geographical locations under the scrutiny of this study (p = 0.003). DISCUSSION AND CONCLUSION: Having attested the marked seroprevalence of BoHV-1, the definitive role of goats in the epidemiology of this virus as a secondary host or reservoir was confirmed by the present study, necessitating the strict monitoring of BoHV-1 in goats by animal health authorities in areas where BoHV-1 abounds in cattle.
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Enfermedades de las Cabras , Cabras , Infecciones por Herpesviridae , Herpesvirus Bovino 1 , Animales , Estudios Seroepidemiológicos , Irán/epidemiología , Enfermedades de las Cabras/epidemiología , Enfermedades de las Cabras/virología , Femenino , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/epidemiología , Infecciones por Herpesviridae/virología , MasculinoRESUMEN
The C-terminal residues of proteins can function as degrons recognized by ubiquitin ligases for proteasomal degradation. Kelch domain-containing protein 3 (KLHDC3) is a substrate receptor for E3 ubiquitin ligase (Cullin2-RING ligase) that targets the C-terminal degrons. UL49.5 is 96 amino-acid type 1 transmembrane protein from bovine herpesvirus 1. Herpesviruses have evolved highly effective strategies to evade the antiviral immune response. One of these strategies is inhibition of the antigen processing and presentation pathway by MHC I, thereby reducing the presentation of the antigenic peptides on the surface of the infected cell. Recently, it has been demonstrated that UL49.5 triggers TAP degradation via recruiting the E3 ubiquitin ligase to TAP. Moreover, the mutagenesis revealed that the mutations within the UL49.5 C-degron sequence (93RGRG96) affect binding of UL49.5 to KLHDC3. In this work the molecular dynamics of KLHDC3 in complexes with the C-terminal decapeptide of the herpesviral protein UL4.95 and its three mutants has been employed to provide a framework for understanding molecular recognition of UL49.5 by KLHDC3. The findings of this study give insights into the interactions of the various degrons with KLHDC3. During the molecular dynamics, an active RGKG mutant adopts a conformation similar to that of the wild type decapeptide, whereas the conformations of two inactive mutants, KGRG and RGRD are significantly different. Both R93K and G96D mutations impair the interactions of the C-terminal glycine with KLHDC3. The findings of this study expand the existing knowledge about the mechanism of protein recognition by Cullin2-RING ligases thus contributing to the design of antiviral and anticancer drugs that can selectively promote or inhibit degradation of the proteins of interest.
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Simulación de Dinámica Molecular , Mutación , Herpesvirus Bovino 1/metabolismo , Herpesvirus Bovino 1/genética , Proteínas Virales/metabolismo , Proteínas Virales/química , Humanos , Degrones , Proteínas del Envoltorio ViralRESUMEN
Bovine alphaherpesvirus type 1 (BoAHV-1) infections lead to compromised herd health and significantly reduced productivity of affected cattle. While BoAHV-1 may cause rhinotracheitis, conjunctivitis, genital infections, and abortions, respiratory tract infections constitute the predominant clinical disease. Immune suppression induced by BoAHV-1 may contribute to co-infections initiating the bovine respiratory disease complex. In this review, the emphasis is to recapitulate the biology and the vaccine technologies currently in use and in development for BoAHV-1, and to discuss the major limitations. Studies on the life cycle and host interactions of BoAHV-1 have resulted in the identification of virulence factors. While several vaccine types, such as vectored vaccines and subunit vaccines, are under investigation, modified live and inactivated BoAHV-1 vaccines are still most frequently used in most areas of the world, whereas attenuated and inactivated marker vaccines are in use in Europe. The knowledge gained from studies on the biology of BoAHV-1 can form a basis for the rational design of future vaccines.
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Enfermedades de los Bovinos , Animales , Bovinos , Enfermedades de los Bovinos/prevención & control , Enfermedades de los Bovinos/virología , Enfermedades de los Bovinos/inmunología , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/prevención & control , Infecciones por Herpesviridae/virología , Infecciones por Herpesviridae/inmunología , Vacunas Virales/inmunología , Herpesvirus Bovino 1/inmunología , Desarrollo de Vacunas , Vacunas Atenuadas/inmunologíaRESUMEN
Bovine alphaherpesvirus 1 (BoHV-1) infection causes respiratory tract disorders and immune suppression and may induce bacterial pneumonia. BoHV-1 establishes lifelong latency in sensory neurons after acute infection. Reactivation from latency consistently occurs following stress or intravenous injection of the synthetic corticosteroid dexamethasone (DEX), which mimics stress. The immediate early transcription unit 1 (IEtu1) promoter drives expression of infected cell protein 0 (bICP0) and bICP4, two viral transcriptional regulators necessary for productive infection and reactivation from latency. The IEtu1 promoter contains two glucocorticoid receptor (GR) responsive elements (GREs) that are transactivated by activated GR. GC-rich motifs, including consensus binding sites for specificity protein 1 (Sp1), are in the IEtu1 promoter sequences. E2F family members bind a consensus sequence (TTTCCCGC) and certain specificity protein 1 (Sp1) sites. Consequently, we hypothesized that certain E2F family members activate IEtu1 promoter activity. DEX treatment of latently infected calves increased the number of E2F2+ TG neurons. GR and E2F2, but not E2F1, E2F3a, or E2F3b, cooperatively transactivate a 436-bp cis-regulatory module in the IEtu1 promoter that contains both GREs. A luciferase reporter construct containing a 222-bp fragment downstream of the GREs was transactivated by E2F2 unless two adjacent Sp1 binding sites were mutated. Chromatin immunoprecipitation studies revealed that E2F2 occupied IEtu1 promoter sequences when the BoHV-1 genome was transfected into mouse neuroblastoma (Neuro-2A) or monkey kidney (CV-1) cells. In summary, these findings revealed that GR and E2F2 cooperatively transactivate IEtu1 promoter activity, which is predicted to influence the early stages of BoHV-1 reactivation from latency. IMPORTANCE: Bovine alpha-herpesvirus 1 (BoHV-1) acute infection in cattle leads to establishment of latency in sensory neurons in the trigeminal ganglia (TG). A synthetic corticosteroid dexamethasone consistently initiates BoHV-1 reactivation in latently infected calves. The BoHV-1 immediate early transcription unit 1 (IEtu1) promoter regulates expression of infected cell protein 0 (bICP0) and bICP4, two viral transcriptional regulators. Hence, the IEtu1 promoter must be activated for the reactivation to occur. The number of TG neurons expressing E2F2, a transcription factor and cell cycle regulator, increased during early stages of reactivation from latency. The glucocorticoid receptor (GR) and E2F2, but not E2F1, E2F3a, or E2F3b, cooperatively transactivated a 436-bp cis-regulatory module (CRM) in the IEtu1 promoter that contains two GR responsive elements. Chromatin immunoprecipitation studies revealed that E2F2 occupies IEtu1 promoter sequences in cultured cells. GR and E2F2 mediate cooperative transactivation of IEtu1 promoter activity, which is predicted to stimulate viral replication following stressful stimuli.
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Ciclo Celular , Factor de Transcripción E2F2 , Regulación Viral de la Expresión Génica , Herpesvirus Bovino 1 , Proteínas Inmediatas-Precoces , Regiones Promotoras Genéticas , Receptores de Glucocorticoides , Activación Transcripcional , Animales , Bovinos , Ratones , Sitios de Unión , Línea Celular , Dexametasona/farmacología , Factor de Transcripción E2F2/metabolismo , Regulación Viral de la Expresión Génica/efectos de los fármacos , Regulación Viral de la Expresión Génica/genética , Infecciones por Herpesviridae/virología , Infecciones por Herpesviridae/metabolismo , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/genética , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/fisiología , Proteínas Inmediatas-Precoces/genética , Neuronas/virología , Receptores de Glucocorticoides/metabolismo , Elementos de Respuesta/genética , Factor de Transcripción Sp1/metabolismo , Transactivadores/metabolismo , Ganglio del Trigémino/citología , Ganglio del Trigémino/virología , Activación Viral , Latencia del VirusRESUMEN
Bovine viral diarrhea virus (BVDV) is considered to be the most common agent of severe diarrhea in cattle worldwide, causing fever, diarrhea, ulcers, and abortion. Bovine herpesvirus 1 (BoHV-1) is also a major bovine respiratory disease agent that spreads worldwide and causes extensive damage to the livestock industry. Recombinase polymerase amplification (RPA) is a novel nucleic acid amplification method with the advantages of high efficiency, rapidity and sensitivity, which has been widely used in the diagnosis of infectious diseases. A dual RPA assay was developed for the simultaneous detection of BVDV and BoHV-1. The assay was completed at a constant temperature of 37 °C for 30 min. It was highly sensitive and had no cross-reactivity with other common bovine viruses. The detection rate of BVDV RPA in clinical samples (36.67%) was higher than that of PCR (33.33%), the detection rate of BoHV-1 RPA and PCR were equal. Therefore, the established dual RPA assay for BVDV and BoHV-1 could be a potential candidate for use as an immediate diagnostic.
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Virus de la Diarrea Viral Bovina , Herpesvirus Bovino 1 , Técnicas de Amplificación de Ácido Nucleico , Recombinasas , Animales , Bovinos , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , Recombinasas/metabolismo , Virus de la Diarrea Viral Bovina/genética , Virus de la Diarrea Viral Bovina/aislamiento & purificación , Sensibilidad y Especificidad , Diarrea Mucosa Bovina Viral/virología , Diarrea Mucosa Bovina Viral/diagnóstico , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/virología , Infecciones por Herpesviridae/diagnóstico , ADN Viral/genéticaRESUMEN
Bovine alphaherpesvirus 1 (BoAHV-1) is the most important respiratory and reproductive disease-causing pathogen in dairy cattle. Despite BoAHV-1 has become widespread and a major challenge to the dairy industry, little is known about its epidemiology in dairy herds in Ethiopia. A cross-sectional study was conducted from November 2022 to May 2023 to determine the seroprevalence and potential risk factors associated with BoAHV-1 seropositivity in dairy herds in North Shewa, the central highlands of Ethiopia. A total of 511 blood samples were collected from randomly selected cattle herds (n = 142) and examined antibodies against BoAHV-1 using ELISA test. A retrospective survey was also done to gather information related to reproductive disorders. The overall seroprevalence of BoAHV-1 was 61.84% (95% CI: 57.53-65.97) at the animal level and 85.21% (95% CI: 78.28-90.21) at the herd level. Multivariable logistic analysis revealed that the risk of being BoAHV-1 seropositive was nine times higher in cows older than six years (OR = 9.16; 95% CI: 3.09-27.16; P = 0.000), five times higher (OR = 4.51; 95% CI: 1.23-16.53; P = 0.019) in cows with a history of abortion, three times higher (OR = 2.75; 95% CI: 1.72-4.22; P = 0.029) in cows with a history of retained fetal membrane, and three times higher (OR = 2.83; 1.86-9.31; P = 0.03) in animals with clinical signs of ocular and/or nasal discharge. This study demonstrates a significant circulating of BoAHV-1 in the dairy cattle population in study districts. Thus, a comprehensive approach that includes strict farm biosecurity and vaccination should be practiced for effective BoAHV-1 control and prevention and to promote the growing dairy industry in the central highlands of Ethiopia.
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Enfermedades de los Bovinos , Infecciones por Herpesviridae , Animales , Bovinos , Etiopía/epidemiología , Estudios Seroepidemiológicos , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/virología , Estudios Transversales , Femenino , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/epidemiología , Infecciones por Herpesviridae/virología , Industria Lechera , Factores de Riesgo , Estudios Retrospectivos , Anticuerpos Antivirales/sangre , Herpesvirus Bovino 1/inmunologíaRESUMEN
Bovine respiratory disease (BRD) is one of the most common diseases in the cattle industry worldwide; it is caused by multiple bacterial or viral coinfections, of which Mycoplasma bovis (M. bovis) and bovine herpesvirus type 1 (BoHV-1) are the most notable pathogens. Although live vaccines have demonstrated better efficacy against BRD induced by both pathogens, there are no combined live and marker vaccines. Therefore, we developed an attenuated and marker M. bovis-BoHV-1 combined vaccine based on the M. bovis HB150 and BoHV-1 gG-/tk- strain previously constructed in our lab and evaluated in rabbits. This study aimed to further evaluate its safety and protective efficacy in cattle using different antigen ratios. After immunization, all vaccinated cattle had a normal rectal temperature and mental status without respiratory symptoms. CD4+, CD8+, and CD19+ cells significantly increased in immunized cattle and induced higher humoral and cellular immune responses, and the expression of key cytokines such as IL-4, IL-12, TNF-α, and IFN-γ can be promoted after vaccination. The 1.0 × 108 CFU of M. bovis HB150 and 1.0 × 106 TCID50 BoHV-1 gG-/tk- combined strain elicited the most antibodies while significantly increasing IgG and cellular immunity after challenge. In conclusion, the M. bovis HB150 and BoHV-1 gG-/tk- combined strain was clinically safe and protective in calves; the mix of 1.0 × 108 CFU of M. bovis HB150 and 1.0 × 106 TCID50 BoHV-1 gG-/tk- strain was most promising due to its low amount of shedding and highest humoral and cellular immune responses compared with others. This study introduces an M. bovis-BoHV-1 combined vaccine for application in the cattle industry.
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Herpesvirus Bovino 1 , Mycoplasma bovis , Vacunas Atenuadas , Vacunas Combinadas , Animales , Bovinos , Herpesvirus Bovino 1/inmunología , Vacunas Combinadas/inmunología , Vacunas Combinadas/administración & dosificación , Vacunas Atenuadas/inmunología , Vacunas Atenuadas/administración & dosificación , Mycoplasma bovis/inmunología , Vacunas Virales/inmunología , Vacunas Virales/administración & dosificación , Vacunas Virales/efectos adversos , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/efectos adversos , Citocinas/metabolismo , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Anticuerpos Antibacterianos/sangre , Anticuerpos Antibacterianos/inmunología , Infecciones por Mycoplasma/prevención & control , Infecciones por Mycoplasma/veterinaria , Infecciones por Mycoplasma/inmunología , Vacunas Marcadoras/inmunología , Vacunas Marcadoras/administración & dosificación , Vacunación/veterinaria , Eficacia de las Vacunas , Inmunidad Humoral , Complejo Respiratorio Bovino/prevención & control , Complejo Respiratorio Bovino/inmunología , Complejo Respiratorio Bovino/virologíaRESUMEN
OBJECTIVE: Compare immune responses induced by 2 commercial intranasal (IN) modified-live viral (MLV) vaccines given individually or coadministered and evaluate prevention of infection and lung pathology following bovine herpesvirus-1 (BHV-1) challenge. ANIMALS: 36 male Holstein calves (ages, 5 to 12 days). METHODS: In a randomized complete block design, each calf received an IN injection of either vaccine diluent (Placebo), an MLV vaccine containing bovine herpesvirus-1 (BHV-1; N3), bovine coronavirus vaccine (BC), or both N3 and BC (BC + N3) with a booster 4 weeks later. Nasal secretions and blood were collected weekly. Three weeks after the booster, the calves were challenged with BHV-1, sampled for virus shedding, and euthanized 10 days later to quantify lung pathology. The study period was September 7, 2020, to April 6, 2021. RESULTS: Calves were seropositive for BHV-1 and BC before vaccination. No significant difference in BC-specific serum immunoglobin G and nasal immunoglobin A antibody responses in the BC versus BC + N3 group or BHV-1-specific serum immunoglobin G and nasal immunoglobin A antibody responses in the N3 versus BC + N3 group. Cytokine responses to BHV-1 and BC did not differ among groups. BHV-1 shedding after challenge was significantly reduced in N3 groups versus Placebo and BC. There was a significant reduction in lung pathology in the N3 + BC group versus Placebo. CLINICAL RELEVANCE: This study provides evidence an MLV vaccine containing BHV-1 and an MLV BC vaccine can be coadministered to neonatal calves without significantly altering immune responses to the 2 viruses or compromising the prevention of BHV-1 respiratory disease. Calves receiving the BC + N3 vaccine had a significant reduction in lung pathology after BHV-1 aerosol challenge.
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Administración Intranasal , Animales Recién Nacidos , Enfermedades de los Bovinos , Infecciones por Coronavirus , Coronavirus Bovino , Infecciones por Herpesviridae , Herpesvirus Bovino 1 , Vacunas Atenuadas , Vacunas Virales , Animales , Bovinos , Herpesvirus Bovino 1/inmunología , Administración Intranasal/veterinaria , Masculino , Vacunas Virales/inmunología , Vacunas Virales/administración & dosificación , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/inmunología , Coronavirus Bovino/inmunología , Enfermedades de los Bovinos/prevención & control , Enfermedades de los Bovinos/virología , Enfermedades de los Bovinos/inmunología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/virología , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/prevención & control , Infecciones por Herpesviridae/inmunología , Infecciones por Herpesviridae/virología , Rinotraqueítis Infecciosa Bovina/prevención & control , Rinotraqueítis Infecciosa Bovina/inmunología , Esparcimiento de Virus , Anticuerpos Antivirales/sangre , Distribución AleatoriaRESUMEN
Probiotics are live microorganisms that, confer health benefits to the host when supplemented in adequate amounts. They can promote immunomodulation by inducing phagocyte activity, leukocyte proliferation, antibody production, and cytokine expression. Lactic acid bacteria (BAL) are important probiotic specimens with properties that can improves ruminant nutrition, productivity and immunity. The aim of the present study was to evaluate the immunomodulatory effect of the supplementation with Lacticaseibacillus casei CB054 in calve vaccinated against bovine infectious rhinotracheitis (IBR). Calve were vaccinated with a commercial IBR vaccine, on day 0 and received a booster dose on day 21. L. casei CB054 was orally administered (4 ×109 UFC) for 35 days, while a non-supplemented control group received Phosphate Buffer Saline (PBS). Stimulation of bovine splenocytes with L. casei CB054 markedly enhanced mRNA transcription levels of cytokines IL2, IL4, IL10 and IL17 genes. Calves supplemented with L. casei CB054 showed significantly higher (p < 0.05) specific anti-BoHV-1 IgG levels, higher serum neutralization, as well as higher mRNA transcription for IL2, IL4, IL10 and IL17 genes in Peripheral Blood Mononuclear Cells (PBMCs) comparing with control calves. Supplemented calve had an average weight gain of â¼14 kg more than non-supplemented during the experimental period. These results suggest that L. casei CB054 supplementation increase immunogenicity of a commercial IBR vaccine in cattle and improve weight gain.
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Enfermedades de los Bovinos , Herpesvirus Bovino 1 , Rinotraqueítis Infecciosa Bovina , Lacticaseibacillus casei , Vacunas , Animales , Bovinos , Interleucina-10 , Interleucina-2 , Interleucina-4 , Leucocitos Mononucleares , Citocinas , Suplementos Dietéticos , Inmunomodulación , Aumento de Peso , ARN Mensajero , Enfermedades de los Bovinos/prevención & controlRESUMEN
Using marker vaccines to control bovine alphaherpesvirus-1 (BoHV-1) is a novel strategy for differentiation between infected and vaccinated animals (DIVA). In this study, multiplex real-time PCR targeting gD and gE genes was applied for BoHV-1 screening on 60 clinical samples from cattle with a history of vaccination, in some cases by US2-deleted marker vaccines, that were suffering from severe respiratory symptoms. Conventional PCR targeting the gC and US2 flanking region was done for molecular characterization and identification of the US2-deleted vaccine strain. Six samples were positive for BoHV-1 by both RT-PCR and conventional PCR. Surprisingly, a conventional PCR DIVA trial based on the US2 gene revealed that only one sample that exhibited the US2 gene was a wild virus, while others that did not exhibit the US2 gene were vaccine viruses. Phylogenetic characterization classifies the samples as BoHV-1.1. This finding reveals the circulation of vaccine virus in field-diseased animals, which threatens the eradication program.
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Enfermedades de los Bovinos , Infecciones por Herpesviridae , Herpesvirus Bovino 1 , Animales , Bovinos , Herpesvirus Bovino 1/genética , Vacunas Marcadoras/genética , Egipto/epidemiología , Filogenia , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
OBJECTIVE: To determine the efficacy of primary or booster intranasal vaccination of beef steers on clinical protection and pathogen detection following simultaneous challenge with bovine respiratory syncytial virus and bovine herpes virus 1. METHODS: 30 beef steers were randomly allocated to 3 different treatment groups starting at 2 months of age. Group A (n = 10) was administered a single dose of a parenteral modified-live vaccine and was moved to a separate pasture. Groups B (n = 10) and C (10) remained unvaccinated. At 6 months of age, all steers were weaned and transported. Subsequently, groups A and B received a single dose of an intranasal modified-live vaccine vaccine while group C remained unvaccinated. Group C was housed separately until challenge. Two days following vaccination, all steers were challenged with bovine respiratory syncytial virus and bovine herpes virus 1 and housed in a single pen. Clinical and antibody response outcomes and the presence of nasal pathogens were evaluated. RESULTS: The odds of clinical disease were lower in group A compared with group C on day 7 postchallenge; however, antibody responses and pathogen detection were not significantly different between groups before and following viral challenge. All calves remained negative for Histophilus somni and Mycoplasma bovis; however, significantly greater loads of Mannheimia haemolytica and Pasteurella multocida were detected on day 7 postchallenge compared with day -2 prechallenge. CLINICAL RELEVANCE: Intranasal booster vaccination of beef steers at 6 months of age reduced clinical disease early after viral challenge. Weaning, transport, and viral infection promoted increased detection rates of M haemolytica and P multocida regardless of vaccination status.
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Administración Intranasal , Coinfección , Herpesvirus Bovino 1 , Inmunización Secundaria , Infecciones por Virus Sincitial Respiratorio , Virus Sincitial Respiratorio Bovino , Animales , Bovinos , Herpesvirus Bovino 1/inmunología , Masculino , Administración Intranasal/veterinaria , Virus Sincitial Respiratorio Bovino/inmunología , Inmunización Secundaria/veterinaria , Coinfección/veterinaria , Coinfección/prevención & control , Coinfección/microbiología , Infecciones por Virus Sincitial Respiratorio/veterinaria , Infecciones por Virus Sincitial Respiratorio/prevención & control , Rinotraqueítis Infecciosa Bovina/prevención & control , Rinotraqueítis Infecciosa Bovina/inmunología , Enfermedades de los Bovinos/prevención & control , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/virología , Enfermedades de los Bovinos/inmunología , Vacunas Virales/inmunología , Vacunas Virales/administración & dosificación , Derrame de Bacterias , Anticuerpos Antivirales/sangre , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/prevención & control , Distribución Aleatoria , Vacunación/veterinariaRESUMEN
The CRISPR/Cas9 system is widely used to manipulate viral genomes. Although Alphaherpesvirinae genomes are large and complicated to edit, in recent years several Pseudorabies virus (PRV) mutants have been successfully generated using the CRISPR/Cas9 system. However, the application of CRISPR/Cas9 editing on another member of alpha herpesviruses, bovine herpesvirus-1 (BHV-1), is rarely reported. This paper reports a rapid and straightforward approach to manipulating herpesviruses genome using CRISPR/Cas9. The recombinant plasmids contained the left and right arm of the thymidine kinase (TK) gene of PRV or of the glycoprotein I (gI) and glycoprotein E (gE) of BHV-1. Upon the cleavage of the TK or gIgE gene by Cas9 protein, this was replaced by the enhanced green fluorescence protein (eGFP) by homologous recombination. With this approach, we generated recombinant TK-/eGFP+ PRV and gIgE-/eGFP+ BHV-1 mutants and then proceeded to characterize their biological activities in vitro and in vivo. In conclusion, we showed that alpha herpesvirus, including PRV and BHV-1, can be rapidly edited using the CRISPR/Cas9 approach paving the way to the development of animal herpesvirus vaccines.
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Herpesvirus Bovino 1 , Herpesvirus Suido 1 , Seudorrabia , Animales , Edición Génica , Sistemas CRISPR-Cas , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/metabolismo , Seudorrabia/prevención & control , Glicoproteínas/genéticaRESUMEN
BACKGROUND: Studies have confirmed that Infectious bovine rhinotracheitis virus (IBRV) infection induces mitochondrial damage. MicroRNAs (miRNAs) are a class of noncoding RNA molecules, which are involved in various biological processes and pathological changes associated with mitochondrial damage. It is currently unclear whether miRNAs participate in IBRV-induced mitochondrial damage in Madin-Darby bovine kidney (MDBK) cells. RESULTS: In the present study, we used high-throughput sequencing technology, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis to screen for mitochondria-related miRNAs and messenger RNAs (mRNAs). In total, 279 differentially expressed miRNAs and 832 differentially expressed mRNAs were identified in 6 hours (IBRV1) versus 24 hours (IBRV2) after IBRV infection in MDBK cells. GO and KEGG enrichment analysis revealed that 42 differentially expressed mRNAs and 348 target genes of differentially expressed miRNAs were correlated with mitochondrial damage, and the miRNA-mitochondria-related target genes regulatory network was constructed to elucidate their potential regulatory relationships. Among the 10 differentially expressed miRNAs, 8 showed expression patterns consistent with the high-throughput sequencing results. Functional validation results showed that overexpression of miR-10a and miR-182 aggravated mitochondrial damage, while inhibition of miR-10a and miR-182 alleviated mitochondrial damage. CONCLUSIONS: This study not only revealed the expression changes of miRNAs and mRNAs in IBRV-infected MDBK cells, but also revealed possible biological regulatory relationship between them. MiR-10a and miR-182 may have the potential to be developed as biomarkers for the diagnosis and treatment of IBRV. Together, Together, these data and analyses provide additional insights into the roles of miRNA and mRNA in IBRV-induced mitochondria damage.
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Herpesvirus Bovino 1 , MicroARNs , Animales , Bovinos , MicroARNs/genética , MicroARNs/metabolismo , Herpesvirus Bovino 1/genética , Células Epiteliales/metabolismo , Riñón/metabolismo , Redes Reguladoras de Genes , ARN Mensajero/genética , Perfilación de la Expresión GénicaRESUMEN
Bovine herpesvirus 1 (BoHV-1) is a highly contagious pathogen which causes infectious bovine rhinotracheitis in cattle worldwide. Although it has the ability to evade the host's antiviral innate immune response and establish persistent latent infections, the mechanisms are not fully understood, especially the function of the tegument protein to escape innate immunity and participate in viral replication. In this study, we showed that overexpression of tegument protein UL3 facilitates BoHV-1 replication and suppresses the expression of type-I interferon (IFN-I) and IFN-stimulated genes. Then, STING was identified as the target by which UL3 inhibits the IFN-I signaling pathway, and STING was degraded through the UL3-induced autophagy pathway. Furthermore, overexpression of UL3 promotes the expression of the autophagy-related protein ATG101, thereby inducing autophagy. Further study showed that UL3 enhances the interaction between ATG101 and STING, and then the degradation of STING was reversed following ATG101 silencing in UL3-overexpressing cells during BoHV-1 infection. Our research results demonstrate a novel function of UL3 in regulating host's antiviral response and provide a potential mechanism for BoHV-1 immune evasion.
Asunto(s)
Infecciones por Herpesviridae , Herpesvirus Bovino 1 , Proteínas Virales , Animales , Bovinos , Antivirales , Autofagia , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/metabolismo , Inmunidad Innata/genética , Replicación Viral/genética , Interferón Tipo I/metabolismo , Proteínas de la Membrana/metabolismo , Infecciones por Herpesviridae/metabolismo , Infecciones por Herpesviridae/veterinaria , Proteínas Virales/metabolismoRESUMEN
There is sufficient evidence that both bovine herpesvirus (BoHV-1) and bubaline herpesvirus (BuHV-1) can overcome the species barrier represented by their respective hosts, cattle and buffalo. Although several studies have focused on the impact of BoHV-1 on buffalo, little is known about the impact of BuHV-1 on cattle. In this work, we evaluated the seroprevalence of BuHV-1 in the cattle population in an area where intensive buffalo farming is highly developed (Campania region, Italy). BuHV-1 seroprevalence of cattle sampled in this study was estimated to be 21.4% using a specific commercial ELISA for the detection of antibodies against glycoprotein E of the virus. Risk factor assessment by univariate analysis revealed a correlation between housing type and higher prevalence. Similarly, cattle housed with buffalo and adult animals had a higher likelihood of being seropositive. BoHV-1 vaccination did not prove to be a protective factor against BuHV-1 exposure. The role of age, grazing, and co-living with buffalo in influencing BuHV-1 exposure was also confirmed by multivariate analysis. All BuHV-1 positive animals were also tested with cross-serum neutralization aimed at evaluating the specific antibody titers against BoHV-1 and BuHV-1. We, therefore, assessed the potential cross-reaction between BoHV-1 and BuHV-1, the co-infection rate, and the agreement of the assays used. This study described the presence of BuHV-1 in the cattle population of the Campania region (Italy) and indicated the requirement to take BuHV-1 into consideration for any measures and control and/or eradication plans to be applied against BoHV-1.
Asunto(s)
Bison , Enfermedades de los Bovinos , Infecciones por Herpesviridae , Herpesviridae , Herpesvirus Bovino 1 , Animales , Bovinos , Infecciones por Herpesviridae/epidemiología , Infecciones por Herpesviridae/veterinaria , Búfalos , Estudios Seroepidemiológicos , Enfermedades de los Bovinos/epidemiología , Anticuerpos AntiviralesRESUMEN
Surveillance of endemic pathogens is essential for disease control, providing an evidence base for policy and advice. Bovine Herpes Virus Type 1 (BoHV-1), the causative agent of Infectious Bovine Rhinotracheitis (IBR), has been found to have high seroprevalence within the Irish cattle population. The aim of the present study was to establish seroprevalence levels for culled cattle in Ireland aged < 30 months and to establish whether BVD exposure and other factors was associated with BoHV-1 exposure. We employed random effects logit models coupled with repeated bootstrap sampling to provide robust estimates. The final dataset contained results for 5273 animals tested over two study years, 2018 and 2020. The animal-level seroprevalence of BoHV-1 was 21.43% (1130/5273; 95%CI: 20.32-22.53%). Univariable analysis suggested that BoHV-1 seropositivity risk was associated with BVDV serodiagnosis status, age, sex, year sampled, herd type, herd-size, and metrics of movement into the herd. Final random-effects multivariable models suggested increased risk associated with increasing herd size of the last herd, movements made by animals during the previous year, and the year the animal was sampled. Despite BVDV status and sex being retained in the final model, repeated bootstrap sampling of the regression model to estimate biased-corrected 95%CI suggested that these associations were not robust. The overall apparent prevalence of BoHV-1 exposure for culled cattle in Ireland declined in 2020 relative to 2018 (from 23.32 to 17.61%). Herd-size and the movement of animals were found to be important factors associated with animal-level risk, but there was less statistical support for sex-based or BVDV status associations.
Asunto(s)
Coinfección , Herpesvirus Bovino 1 , Infecciones por Pestivirus , Animales , Bovinos , Irlanda/epidemiología , Estudios Seroepidemiológicos , Factores de Riesgo , DiarreaRESUMEN
Bovine herpesvirus type 1 (BoHV-1) is a pathogen of cattle responsible for infectious bovine rhinotracheitis. The BoHV-1 UL49.5 is a transmembrane protein that binds to the transporter associated with antigen processing (TAP) and downregulates cell surface expression of the antigenic peptide complexes with the major histocompatibility complex class I (MHC-I). KLHDC3 is a kelch domain-containing protein 3 and a substrate receptor of a cullin2-RING (CRL2) E3 ubiquitin ligase. Recently, it has been identified that CRL2KLHDC3 is responsible for UL49.5-triggered TAP degradation via a C-degron pathway and the presence of the degron sequence does not lead to the degradation of UL49.5 itself. The molecular modeling of KLHDC3 in complexes with four UL49.5 C-terminal decapeptides (one native protein and three mutants) revealed their activity to be closely correlated with the conformation which they adopt in KLHDC3 binding cleft. To analyze the interaction between UL49.5 and KLHDC3 in detail, in this work a total of 3.6 µs long molecular dynamics simulations have been performed. The complete UL49.5-KLHDC3 complexes were embedded into the fully hydrated all-atom lipid membrane model with explicit water molecules. The network of polar interactions has been proposed to be responsible for the recognition and binding of the degron in KLHDC3. The interaction network within the binding pocket appeared to be very similar between two CRL2 substrate receptors: KLHDC3 and KLHDC2.
Asunto(s)
Herpesvirus Bovino 1 , Proteínas del Envoltorio Viral , Animales , Bovinos , Proteínas del Envoltorio Viral/química , Ligasas/metabolismo , Ubiquitina/metabolismo , Degrones , Herpesvirus Bovino 1/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Ubiquitina-Proteína LigasasRESUMEN
Bovine alphaherpesvirus 1 (BoHV-1) infections cause respiratory tract disorders and suppress immune responses, which can culminate in bacterial pneumonia. Following acute infection, BoHV-1 establishes lifelong latency in sensory neurons present in trigeminal ganglia (TG) and unknown cells in pharyngeal tonsil. Latently infected calves consistently reactivate from latency after an intravenous injection of the synthetic corticosteroid dexamethasone (DEX), which mimics the effects of stress. The immediate early transcription unit 1 (IEtu1) promoter drives expression of infected cell protein 0 (bICP0) and bICP4, two key viral transcriptional regulators. The IEtu1 promoter contains two functional glucocorticoid receptor (GR) response elements (GREs), and this promoter is transactivated by GR, DEX, and certain Krüppel transcription factors that interact with GC-rich motifs, including consensus specificity protein 1 (Sp1) binding sites. Based on these observations, we hypothesized that Sp1 stimulates productive infection and transactivates key BoHV-1 promoters. DEX treatment of latently infected calves increased the number of Sp1+ TG neurons and cells in pharyngeal tonsil indicating that Sp1 expression is induced by stress. Silencing Sp1 protein expression with siRNA or mithramycin A, a drug that preferentially binds GC-rich DNA, significantly reduced BoHV-1 replication. Moreover, BoHV-1 infection of permissive cells increased Sp1 steady-state protein levels. In transient transfection studies, GR and Sp1 cooperatively transactivate IEtu1 promoter activity unless both GREs are mutated. Co-immunoprecipitation studies revealed that GR and Sp1 interact in mouse neuroblastoma cells (Neuro-2A) suggesting this interaction stimulates IEtu1 promoter activity. Collectively, these studies suggested that the cellular transcription factor Sp1 enhances productive infection and stress-induced BoHV-1 reactivation from latency.IMPORTANCEFollowing acute infection, bovine alphaherpesvirus 1 (BoHV-1) establishes lifelong latency in sensory neurons in trigeminal ganglia (TG) and pharyngeal tonsil. The synthetic corticosteroid dexamethasone consistently induces BoHV-1 reactivation from latency. The number of TG neurons and cells in pharyngeal tonsil expressing the cellular transcription factor specificity protein 1 (Sp1) protein increases during early stages of dexamethasone-induced reactivation from latency. Silencing Sp1 expression impairs BoHV-1 replication in permissive cells. Interestingly, mithramycin A, a neuroprotective antibiotic that preferentially binds GC-rich DNA, impairs Sp1 functions and reduces BoHV-1 replication suggesting that it is a potential antiviral drug. The glucocorticoid receptor (GR) and Sp1 cooperatively transactivate the BoHV-1 immediate early transcript unit 1 (IEtu1) promoter, which drives expression of infected cell protein 0 (bICP0) and bICP4. Mithramycin A also reduced Sp1- and GR-mediated transactivation of the IEtu1 promoter. These studies revealed that GR and Sp1 trigger viral gene expression and replication following stressful stimuli.
Asunto(s)
Infecciones por Herpesviridae , Herpesvirus Bovino 1 , Receptores de Glucocorticoides , Factor de Transcripción Sp1 , Animales , Bovinos , Ratones , Corticoesteroides/metabolismo , Dexametasona/farmacología , ADN/metabolismo , Herpesvirus Bovino 1/fisiología , Plicamicina/análogos & derivados , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismo , Factores de Transcripción/metabolismo , Proteínas Virales/metabolismo , Factor de Transcripción Sp1/metabolismoRESUMEN
Bovine herpesvirus 1(BoHV-1) is an important bovine pathogen that causes great economic loss to cattle farms worldwide. The virus-productive infection in bovine kidney (MDBK) cells results in ATP depletion. The mechanisms are not well understood. Mitochondrial fatty acid ß-oxidation (FAO) is an important energy source in many tissues with high energy demand. Since carnitine palmitoyl-transferase 1 A (CPT1A) is the rate-limiting enzyme of FAO, we investigated the interactions between virus-productive infection and CPT1A signaling. Here, we found that virus-productive infection at the later stage significantly decreased CPT1A protein levels in all the detected cells, including MDBK, A549, and Neuro-2A cells, differentially altered the accumulation of CPT1A proteins in the nucleus and cytosol, and re-localized the protein in the nucleus. Etomoxir (ETO), an irreversible inhibitor of CPT1A, inhibited viral replication and partially interfered with the ability of BoHV-1 to alter CPT1A accumulation in the nucleus but not in the cytosol. Furthermore, ETO consistently reduced RNA levels of two viral regulatory proteins (bICP0 and bICP22) and protein expression of virion-associated proteins during productive infection, further supporting the important roles of CPT1A signaling in BoHV-1 productive infection. These data, for the first time, suggest that CPT1A is potentially involved in BoHV-1 productive infection.
Asunto(s)
Enfermedades de los Bovinos , Infecciones por Herpesviridae , Herpesvirus Bovino 1 , Bovinos , Animales , Herpesvirus Bovino 1/genética , Replicación Viral , Infecciones por Herpesviridae/veterinaria , Transferasas/metabolismo , Carnitina/metabolismoRESUMEN
Enzootic bovine leukosis virus (BLV) and bovine herpesvirus 1 (BHV-1) are very important infectious agents for the livestock industry worldwide. The present study aimed to explore the association between natural exposure to BLV and BHV-1 with sperm quality analyzed by Computer-Assisted Semen Analysis (CASA) systems. Ten sexually mature Brahman bulls, with sanitary status BLV+/BHV-1+ (n = 2), BLV-/BHV-1+ (n = 6) and BLV-/BHV-1- (n = 2) were evaluated twice, 30 days apart. Results showed that sanitary status of each bull was not associated with semen quality. It was found that the quality of the semen from the second collection was better due to the interruption of sexual rest. The evidence thus revealed that a bull infected with BLV generated good-quality contaminated semen and, therefore, that it is essential to detect contaminated seminal samples to prevent the spread of BLV. A multivariate analysis showed the presence of four sperm subpopulations in Brahman bulls that differ significantly in their kinematic patterns and with respect to sanitary status (P < 0.05), indicating that infection-free and seronegative bulls present the best kinematic parameters, which improved discrimination of sperm quality according to sanitary status. Overall, the analyses indicate that the seropositive-infected bulls with BLV and BHV-1 should be excluded from beef cattle farms.