Bovine Herpesvirus Type Research Articles

A Novel Rabies Vaccine Based on a Recombinant Bovine Herpes Virus Type 1 Expressing Rabies Virus Glycoprotein.

Rabies is a highly prevalent zoonotic disease and a public health threat worldwide. Currently licensed rabies vaccines are effective but less is known which would protect cattle. This study describes the construction of a novel recombinant bovine herpes virus type I (BHV-1) expressing rabies virus glycoprotein (RABV G) instead of its gE glycoprotein (gE) by CRISPR-Cas9 and homologous recombination technology (BHV-1-ΔgE-G). Insertion of the RABV G gene is stable after 20 rounds of in vitro passaging and the recombinant virus replicates to high titers in MDBK cells. The RABV G expresses in the recombinant virus-infected cells and on the virion surface of BHV-1-ΔgE-G. One single immunization with BHV-1-ΔgE-G-activated dendritic cells (DCs) and B cells furthermore induced a protective immune response in mice against severe lethal challenge infection. A protective level of RABV-specific virus-neutralizing antibody (VNA) was detected in intramuscular immunized mice and cattle without any clinical symptoms. This research demonstrated that the BHV-1 vector-based RABV vaccine is a potential candidate for cattle.

Development of differentiation method for bovine herpesvirus serotypes (BHV-1, BHV-4, BHV-5) using polymerase chain reaction

Infectious pneumoenteritis of cattle is etiologically caused by viruses of different families and species. Bovine herpesvirus type 1 — infectious rhinotracheitis virus — is the main and the most dangerous pathogen transmitted by cattle semen. At the same time, recently, according to European scientists’ data, in addition to this pathogen, other herpesviruses have been circulating in cattle groups, in particular, bovine herpesvirused of types 4 and 5. Studies have been conducted using molecular-genetic and bioinformatic methods. Based on the analysis of the genomes of bovine herpesvirus of types 1 (IBR virus), 4 and 5 we developed primers BoHV-1 F/R, which flanks the DNA fragment of the IBR virus with the length of 204 bp, BoHV-4 F/R, which flanks the DNA fragment of bovine herpesvirus type 4 with the length of 615 bp, and BoHV-5 F/R for bovine herpesvirus type 5 DNA amplification with the formation of specific fragments 158 bp in length. The tests demonstrated that primers specific for bovine herpesvirus of types 1, 4 and 5 can be used in multiplex amplification format and hybridized only with specific DNA matrices of bovine herpesviruses. A standard operating procedure ‘Indication of DNA of infectious bovine rhinotracheitis virus and bovine herpesviruses of types 4 and 5 by polymerase chain reaction’ has been developed

Niveles de anticuerpos frente a Herpesvirus Bovino tipo 1 y factores de riesgo asociados en rebaños lecheros no vacunados en un clima húmedo tropical, Ecuador

In order to determine the antibodies levels against Bovine Herpesvirus type 1 (BHV-1) and associated risk factors in unvaccinated dairy herds of tropical wet weather of Ecuador, an investigation was carried out in thirteen production units (PU), in the Chone Canton, Province of Manabí. The sample size was calculate by cluster sampling and the variables to be evaluated were antibodies levels frequency, age, sex, ocular or vulvovaginal lesions, breed and PU. One hundred eighty three blood samples were taken from the jugular or caudal vein and labeled for transfer to the laboratory. The tubes were centrifuged at 3,220 g for 15 minutes and the sera were transferred to Eppendorf tubes that once labeled were stored at –20°C until processing. Antibodies levels against HBV-1 were measured through of competitive ELISA technique, and calculated with simples statistical. The risk factors were estimated through the Odds ratio and relative risk, and the significance level through the Chi-square. The general antibodies levels frequency was 58.47 %. According age, 28.57 % in animals under two years old (yr.old) and 67.38 % for cattle over two yr.old. According to the sex, males showed 18.18 % and females 82.61 %. The determined risk factors (P<0.05) were age and sex, with a significant relative risk (P<0.05) of 3.31 and 2.32, respectively. The unvaccinated dairy herds of Western lowlands of Ecuador, have high antibodies levels frequency against BHV-1, with females over two yr.old at the highest risk of presenting infectious bovine rhinotracheitis (IBR).

Application of the Integrated Supercritical Fluid Extraction–Impregnation Process (SFE-SSI) for Development of Materials with Antiviral Properties

The integrated supercritical fluid extraction–impregnation process (SFE-SSI) was performed to fabricate material with antiviral properties against the herpes simplex virus (HSV). Cotton gauze and starch/chitosan polymer films (SCF) were impregnated with components extracted from Melissa officinalis at 10 MPa and 40 °C using a green medium, supercritical carbon dioxide (scCO2). The influences of the processing mode regarding the flow of the supercritical fluid through the system, and the mass ratio of the plant material and the solid carrier, on the impregnation yield of M. officinalis extract were studied. The results revealed that the introduction of a fresh amount of CO2 into the system enabled the highest impregnation yield of 2.24% for cotton gauze and 8.71% for SCF. The presence of M. officinalis extract on the surface of both impregnated cotton gaze and SCF was confirmed by FTIR and GC analyses after the re-extraction of the impregnated samples. The M. officinalis impregnated materials showed a strong inhibitory effect against Bovine herpesvirus type 1 (BHV-1).

Concurrent Gene Insertion, Deletion, and Inversion during the Construction of a Novel Attenuated BoHV-1 Using CRISPR/Cas9 Genome Editing.

Bovine herpesvirus type I (BoHV-1) is an important pathogen that causes respiratory disease in bovines. The disease is prevalent worldwide, causing huge economic losses to the cattle industry. Gene-deficient vaccines with immunological markers to distinguish them from wild-type infections have become a mainstream in vaccine research and development. In order to knock out the gE gene BoHV-1, we employed the CRISPR/Cas9 system. Interesting phenomena were observed at the single guide RNA (sgRNA) splicing site, including gene insertion, gene deletion, and the inversion of 5′ and 3′ ends of the sgRNA splicing site. In addition to the deletion of the gE gene, the US9 gene, and the non-coding regions of gE and US9, it was found that the US4 sequence, US6 sequence, and part of the US7 sequence were inserted into the EGFP sgRNA splicing site and the 3′ end of the EGFP sequence was deleted. Similar to the BoHV-1 parent, the BoHV-1 mutants induced high neutralizing antibodies titer levels in mice. In summary, we developed a series of recombinant gE-deletion BoHV-1 samples using the CRISPR/Cas9 gene editing system. The mutant viruses with EGFP+ or EGFP− will lay the foundation for research on BoHV-1 and vaccine development in the future.

β-Catenin-Specific Inhibitor, iCRT14, Promotes BoHV-1 Infection-Induced DNA Damage in Human A549 Lung Adenocarcinoma Cells by Enhancing Viral Protein Expression.

Oncolytic bovine herpesvirus type 1 (BoHV-1) infection induces DNA damage in human lung adenocarcinoma cell line A549. However, the underlying mechanisms are not fully understood. We found that BoHV-1 infection decreased the steady-state protein levels of p53-binding protein 1 (53BP1), which plays a central role in dictating DNA damage repair and maintaining genomic stability. Furthermore, BoHV-1 impaired the formation of 53BP1 foci, suggesting that BoHV-1 inhibits 53BP1-mediated DNA damage repair. Interestingly, BoHV-1 infection redistributed intracellular β-catenin, and iCRT14 (5-[[2,5-Dimethyl-1-(3-pyridinyl)-1H-pyrrol-3-yl]methylene]-3-phenyl-2,4-thiazolidinedione), a β-catenin-specific inhibitor, enhanced certain viral protein expression, such as the envelope glycoproteins gC and gD, and enhanced virus infection-induced DNA damage. Therefore, for the first time, we provide evidence showing that BoHV-1 infection disrupts 53BP1-mediated DNA damage repair and suggest β-catenin as a potential host factor restricting both virus replication and DNA damage in A549 cells.

Limitations of bacterial culture, viral PCR, and tulathromycin susceptibility from upper respiratory tract samples in predicting clinical outcome of tulathromycin control or treatment of bovine respiratory disease in high-risk feeder heifers.

A cross-sectional prospective cohort study including 1026 heifers administered tulathromycin due to high risk of clinical signs of bovine respiratory disease (BRD), measured poor association between BRD clinical outcomes and results of bacterial culture and tulathromycin susceptibility from BRD isolates of deep nasopharyngeal swabs (DNS) and adequate association with viral polymerase chain reaction (PCR) results from nasal swabs. Isolation rates from DNS collected on day-0 and at 1st BRD-treatment respectively were: Mannheimia haemolytica (10.9% & 34.1%); Pasteurella multocida (10.4% & 7.4%); Mycoplasma bovis (1.0% & 36.6%); and Histophilus somni (0.7% & 6.3%). Prevalence of BRD viral nucleic acid on nasal swabs collected exclusively at 1st BRD-treatment were: bovine parainfluenza virus type-3 (bPIV-3) 34.1%; bovine viral diarrhea virus (BVDV) 26.3%; bovine herpes virus type-1 (BHV-1) 10.8%; and bovine respiratory syncytial virus (BRSV) 54.1%. Increased relative risk, at 95% confidence intervals, of 1st BRD-treatment failure was associated with positive viral PCR results: BVDV 1.39 (1.17–1.66), bPIV-3 1.26 (1.06–1.51), BHV-1 1.52 (1.25–1.83), and BRSV 1.35 (1.11–1.63) from nasal swabs collected at 1st BRD-treatment and culture of M. haemolytica 1.23 (1.00–1.51) from DNS collected at day-0. However, in this population of high-risk feeder heifers, the predictive values of susceptible and resistant isolates had inadequate association with BRD clinical outcome. These results indicate, that using tulathromycin susceptibility testing of isolates of M. haemolytica or P. multocida from DNS collected on arrival or at 1st BRD-treatment to evaluate tulathromycin clinical efficacy, is unreliable.

Ocimum sanctum: in vitro antiviral potential against animal viruses

Ocimum sanctum, a well and widely used medicinal plant has been reported to be efficient for many ailments quoted in Ayurvedic literature. It has been reported to treat the various ailments due to viral infections in humans like measles, chickenpox, influenza and Smallpox. Besides, it has also shown its potential to give protection in malignancy related disorders. The present study was carried out to screen its potential against animal viruses infecting the dairy animals. Bovine Herpes Virus-type-1 (BHV-1) and Foot and Mouth disease virus (FMDV) affecting cattles and Newcastle Disease Virus (NDV) affecting poultry were screened. Cytopathic inhibition test was used to check the antiviral effect of O. sanctum against BHV-1 and FMDV in MDBK and BHK cell lines, respectively. Chick embryo fibroblasts were cultured to propagate NDV and tested by haemagglutination inhibition test. The maximum non-toxic dose of Ocimum sanctum leaves was determined in MDBK and BHK cell lines as well as in chick fibroblast by MTT assay. Ocimum sanctum nontoxic concentrations, 2.5 mg/mL, 5 mg/mL and 10 mg/mL were found in MDBK cell line, BHK cell line and chick fibroblast culture respectively. Concentrations lower than MNTD were used for studies. 85.3% and 98.4% protection were recorded against BHV-1 and NDV, respectively. However, no significant effect against FMD virus was observed. Thus, it can be effectively utilised in curing these viral diseases in farm animals.

Antiviral Property of the Fungal Metabolite 3-O-Methylfunicone in Bovine Herpesvirus 1 Infection.

Bovine herpesvirus type-1 (BoHV-1) is a widespread pathogen that provokes infectious rhinotracheitis and polymicrobial infections in cattle, resulting in serious economic losses to the farm animal industry and trade restrictions. To date, non-toxic active drugs against BoHV-1 are not available. The exploitation of bioactive properties of microbial products is of great pharmaceutical interest. In fact, fungi are a promising source of novel drugs with a broad spectrum of activities and functions, including antiviral properties. Hence, the potential antiviral properties of 3-O-methylfunicone (OMF), a secondary metabolite produced by Talaromyces pinophilus, were evaluated on BoHV-1. In this study, during BoHV-1 infection in bovine cells (MDBK), the non-toxic concentration of 5 µM OMF considerably reduced signs of cell death and increased cell proliferation. Furthermore, OMF significantly decreased the virus titer as well as the cytopathic effect and strongly inhibited the expression of bICP0, the major regulatory protein in the BoHV-1 lytic cycle. These findings were accompanied by a considerable up-regulation in the expression of the aryl hydrocarbon receptor (AhR), a multifunctional transcription factor also linked to the host’s response to a herpesvirus infection. Overall, our results suggest that by involving AhR, OMF shows potential against a BoHV-1 infection.

High mortality rate of shipping fever cases in cattle caused by bovine herpesvirus type 1 (BoHV-1)

This study reports the high prevalence and molecular characterization of BoHV-1 infection in imported cattle with respiratory system disease after international transport. A high mortality rate of 14.16% (51/360) was reported in a group of animals imported from Hungary to Turkey in 2019. A total of 17 samples were evaluated (3 lung tissue and 14 nasal swab samples) from 15 cattle aged 6 to 9 months not vaccinated against BoHV-1. Virus isolation, polymerase chain reaction (PCR) and restriction endonuclease analysis (REA) procedures were performed within the scope of this study. By virus isolation in MDBK cells, cytopathologic effects was detected in 8 samples (3 lung tissue and 5 nasal swabs samples). The same eight samples were also found positive by BoHV-1 PCR targeting gC (UL44) gene region. According to the sequencing result, the sample (ID: 10054) dropped into a cluster of BoHV-1.1. The REA was applied to the samples to confirm the results of phylogenetic analysis. All of the isolates were identified in the subgroup BoHV-1.1 by REA. These results showed a high mortality risk for imported animals and the possibility for BoHV-1 entering the receiving country via imported animals after transport. This event is a serious problem both for the control of BoHV-1 as well as for animal health and welfare.

Study of isolate of Infectious rynotracheitis virus identified in the acute course of the disease

Isolation of the virus from biological material from a two-month-old calf with pathology of the respiratory system from a herd with a morbidity rate of 48% was performed. After detection the presence of IRT antigens in the lungs of the dead animal, the pathogen was isolated on a continuous culture of calf kidney cells, where a characteristic cytopathic effect was observed. The genetic material of the bovine herpesvirus type 1 (Bovine herpesvirus-1, BHV-1) was identified by polymerase chain reaction in the test sample. The virus isolate was adapted to continuous cell cultures of calf kidneys, sheep kidney, cow embryo lung and calf trachea, and the most suitable biological system was determined, where adsorption and reproductive properties of the virus were more pronounced. It was found that the highest titer of infectious activity of BHV-1 isolate (6.1 lg TCD50/cm3) was obtained on continuous culture of lung embryonic cells of a cow embryo after its reproduction during three consecutive passages (observation period)

Risk assessment and seroprevalence of bovine herpesvirus type 1 infection in dairy herds in the inner Aegean Region of Turkey

This cross-sectional study was performed to investigate the seroprevalence and associated risk factors of bovine herpesvirus type 1 (BoHV-1) infection in dairy cattle herds in Afyonkarahisar province in the Aegean Region of Turkey. Blood samples were collected from 602 cattle from 56 unvaccinated dairy herds between May 2018 and June 2019. Animal and herd-level epidemiological information was collected with a questionnaire during blood collection. Specific antibodies against BoHV-1 and bovine viral diarrhea virus (BVDV) were detected by using a virus neutralization test and a commercial indirect ELISA kit, respectively. Univariable and multivariable logistic regression analyses were used to determine any association between categorical variables and BoHV-1 seropositivity. The animal-level and herd-level seroprevalences of BoHV-1 infection were determined to be 39.53% (95% confidence interval, CI: 35.71–43.50) and 73.21% (95% CI: 60.41–83.04), respectively. Within-herd prevalence was more than 50% in 34.14% of infected herds. Cattle age (odds ratio, OR= 2.34, 95% CI: 1.58–3.44), BVDV infection (OR= 7.74, 95% CI: 5.08–11.76), and the presence of goats in the herd (OR= 2.84, 95% CI: 1.91–4.19) were identified as risk factors for BoHV-1 seropositivity by the multivariable logistic regression model. This is the first study conducted in Turkey using two-layer sampling and logistic regression analyses to determine the herd-level and animal-level seroprevalence and associated risk factors of BoHV-1 infection.

Parenteral Antioxidant Supplementation at Birth Improves the Response to Intranasal Vaccination in Newborn Dairy Calves.

Newborn calves experience oxidative stress throughout the first month of their life, which is known to decrease lymphocyte functions relevant to vaccine responsiveness. Thus, this study aimed to determine the extent to which parenteral antioxidant supplementation given at birth improves the response to an intranasal viral vaccine in the first month of life of newborn dairy calves. For this, 21 calves were randomly assigned at birth to one of two commercially available antioxidant micronutrient supplements or a placebo group receiving 0.9% sterile saline (n = 7/group). Serum and nasal secretion samples were collected before administration of treatments and an intranasal vaccine against respiratory viruses (bovine herpesvirus type 1, bovine syncytial respiratory virus, and parainfluenza 3), and once weekly for the first four weeks of age. Systemic redox balance was determined in serum. Immunoglobulin A specific for bovine herpesvirus 1 and bovine syncytial respiratory virus was quantified in nasal secretions as a proxy to intranasal vaccine responsiveness. Our results showed that parenteral administration of antioxidants at birth improved calves’ redox balance. Additionally, calves receiving antioxidant supplementation had higher concentrations of immunoglobulin A in their nasal secretions than calves in the control group. Thus, we conclude that supplementation of calves with antioxidants at birth could be a practical strategy to improve intranasal vaccine response. Future larger studies should evaluate the extent to which this increased mucosal response to intranasal vaccination could result in decreased calf morbidity and mortality.

Seroprevalence of Bovine Viral Diarrhoea Virus, Bovine Herpesvirus Type 1 and Bovine Herpesvirus Type 4 infections in cattle in Ağrı Province, Eastern Anatolia Region, Turkey

This research was carried out to determine the seroprevalence of Bovine Viral Diarrhoea Virus (BVDV), Bovine Herpes Virus 1 (BoHV-1), and Bovine Herpes Virus 4 (BoHV-4) in cattle (n = 188) in Ağrı province of Eastern Anatolia Region, Turkey. The presence of BVDV, BoHV-1, and BoHV-4 antibodies in sera samples were determined by an ELISA test kit. It was determined that the highest seropositivity for BoHV-1 in Tutak disrict (9.9%), in ≤ 3 years age groups (3.39%), and in Simmental cattle (14.28%); for BoHV-4 in Doğubeyazıt disrict (4.16%), %), in ≤ 3 years age groups (1.70%), and in Simmental cross breed cattle (4.54%); for BVDV in Diyadin disrict (8.33%), in ≤ 3 years age groups (3.39%), and in Eastern Anatolian Red cattle (11.11%); for BoHV-1 + BoHV-4 in Hamur disrict (14.28%), in ≤ 3 years age groups (10.17%), and in Simmental cattle (14.28%); for BoHV-1 + BVDV in Tutak disrict (30.30%), in 6 > years age groups (18.46%), and in Holstein cross-breed cattle (25%); for BoHV-4 + BVDV in Patnos disrict (12.5%), in ? 3 years age groups (5.08%), and in Holstein cross-breed cattle (25%); for BoHV-1 + BoHV-4 + BVDV in Ağrı center (83.87%), in >3- 6 ≤ years age groups (71.78%), and in Anatolian Black cross-breed cattle (100%) were found in the studied population. Rhinotracheitis and conjunctivitis were determined as the most common clinical signs in seropositive animals. In conclusion, the data obtained in this study showed that exposure to BoHV-1, BVDV and BoHV-4 agents were common among cattle in Ağrı province in the Eastern Anatolia Region of Turkey. Additionally, the highest positivity was determined in BoHV-1 + BoHV-4 + BVDV.

Identification and Genetic Characterization of Viral Pathogens in Ruminant Gestation Abnormalities, Israel, 2015–2019

Infectious agents including viruses are important abortifacients and can cause fetal abnormalities in livestock animals. Here, samples that had been collected in Israel from aborted or malformed ruminant fetuses between 2015 and 2019 were investigated for the presence of the following viruses: the reoviruses bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV), the flaviviruses bovine viral diarrhea virus (BVDV) and border disease virus (BDV), the peribunyaviruses Shuni virus (SHUV) and Akabane virus (AKAV), bovine herpesvirus type 1 (BoHV-1) and bovine ephemeral fever virus (BEFV). Domestic (cattle, sheep, goat) and wild/zoo ruminants were included in the study. The presence of viral nucleic acid or antigen could be confirmed in 21.8 % of abnormal pregnancies (213 out of 976 investigated cases), with peribunyaviruses, reoviruses and pestiviruses being the most prevalent. At least four different BTV serotypes were involved in abnormal courses of pregnancy in Israel. The subtyping of pestiviruses revealed the presence of two BDV and several distinct BVDV type 1 strains. The peribunyaviruses AKAV and SHUV were identified annually throughout the study period, however, variation in the extent of virus circulation could be observed between the years. In 2018, AKAV even represented the most detected pathogen in cases of small domestic ruminant gestation abnormalities. In conclusion, it was shown that various viruses are involved in abnormal courses of pregnancy in ruminants in Israel.

PSIII-25 The effects of varying levels of trace mineral supplementation on mineral balance and antibody concentrations in feedlot cattle

Abstract Trace mineral (TM) supplementation is critical for optimal performance and health. The objective was to determine the effects of varying levels of TM supplementation on feedlot cattle TM status and antibody concentrations. Angus steers (n = 240; BW = 291 kg ± 27.4) were stratified by arrival BW and source and assigned to 1 of 4 treatments (12 pens/treatment; 5 steers/pen). Treatments included a negative control (CON) in which cattle received no TM supplementation, a requirement treatment (1X) in which cattle received added TM at 2016 Nutrient Requirements of Beef Cattle required levels, a 2 times requirements (2X), and a 4 times requirements (4X) treatment. Selenium was included at 0.1, 0.2, and 0.3 mg/kg for 1X, 2X, and 4X respectively. Treatments were chosen based on reported TM supplementation levels from the 2015 Feedlot Consulting Nutritionist Survey. One steer was chosen at random from each pen to be evaluated for serum and liver TM status and antibody concentrations to respiratory viruses. There was treatment × day interaction for serum Co, and liver Cu and Se (P < 0.0001). Serum Co was greatest for the 4X treatment from d 28 through harvest. Liver Cu was greatest for the 2X and 4X treatments from d 56 through harvest. Liver Se was greatest for 2X and 4X from d 28 through harvest. There was an effect of day on liver Co, Fe, Mn, Mo, and Zn (P < 0.0001) and serum Cu, Mn, Mo, Se, and Zn (P < 0.0019). Concentrations for individual TM had different trends over time, however, all reported values were within normal ranges. Serum Zn was greater at harvest (P = .02). There was an effect of time on Bovine Viral Diarrhea Virus Type 1A, Bovine Herpesvirus Type 1, Bovine Parainfluenza 3 virus antibody titer concentrations (P < 0.0001).

Bovine Herpesvirus Types 5 (BoHV5): A New and Emerging Threat in Indian Cattle

Bovine herpesvirus 5 (BoHV-5) belongs to the genus varicellovirus family herpesviridae and is a causative agent of meningo-encephalitis in cattle. BoHV-5 can replicates in the central nervous system (CNS) to cause encephalitis and establishes latency in the trigeminal ganglion of either naturally infected cattle or experimentally inoculated laboratory animals. BoHV 5 was first time reported in Australia in 1962 and subsequently reported in Latin Ameican countries Argentina, Brazil and Uruguay. Few cases of BoHV5 infection has also been reported in USA and Europe. Recently, the BoHV5 infection was reported in Indian cattle for the first time. The BoHV5 was isolated from aborted cattle and characterized as BoHV5 subtype A and this finding may necessitate inclusion of BoHV5 test protocol in testing of semen for sexually transmitted diseases. Because BoHV5 infection in cattle recently reported in India so, it is needed to explore the similarity of the isolates with earlier reported isolates and to check it’s pathogenicity to cause encephalitis in natural host and in experimental animals like rabbits, rat, guniea pig etc. Beside this, we have to explore the information about co-infection of BoHV5 with other viral, bacterial, fungal as well as parasitic agents.

Isolation and Molecular Characterization of Bovine Herpes Virus Type 1 from Upper Respiratory Tract of Bovines

Background: Bovine herpes virus type 1 (BoHV-1) the causative agent of Infectious bovine rhinotracheitis (IBR) is of great concern to dairy farmers and veterinarians due to great economic impact caused by the virus in terms of loss of production and abortion. Therefore, the study was planned to detect the virus circulating in the bovine population of the region under study. This virus is an important pathogen of bovine respiratory diseases. The aim of the present study was to isolate the BoHV-1 virus from the upper respiratory tract of bovines. Methods: A total of 13 nasal swab samples were subjected to virus isolation in Madin Darby Bovine Kidney (MDBK) cells lines. A PCR assay was applied to confirm the BoHV-1 DNA by targeting gI glycoprotein gene in isolates. Result: Total two IBR virus isolates were recovered from 13 nasal swab samples of bovines. Both isolates exhibited cytopathic effects i.e. clumping and rounding of cells. A 468 base pair of amplified product from both isolates confirmed the IBR virus in gI gene specific PCR for BoHV-1. This study concludes that IBR virus exists among cattle population of Punjab and it is present in the upper respiratory tract of infected animal and shed through respiratory route. The PCR detection assay for detection of BoHV-1 from nasal swab samples is considerably more sensitive than virus isolation.

Innate and acquired immune responses of colostrum-fed neonatal Holstein calves following intranasal vaccination with two commercially available modified-live virus vaccines.

To compare immune responses induced by 2 commercially available vaccines with a bovine herpesvirus type 1 (BHV1) component following intranasal (IN) administration to colostrum-fed calves. 90 male Holstein calves (ages, 5 to 14 days). In a randomized complete block design, each calf received 2 mL (1 mL/nostril) of vaccine A (n = 30), vaccine B (30), or saline (0.9% NaCl) solution (30) on day 0. Blood samples were collected for determination of serum anti-BHV1 IgG titer, and nasal fluid (NF) samples were collected for determination of interferon (IFN)-α and IFN-γ concentrations and for secretory IgA titers against BHV1, Mannheimia haemolytica, and Pasteurella multocida at predetermined times for 42 days after vaccination. All calves were seropositive for anti-BHV1 IgG, and the mean anti-BHV1 IgG titer did not differ significantly among the 3 groups at any time. Both vaccines induced significant transient increases in NF IFN-α and IFN-γ concentrations. On day 5, mean IFN-α concentration and the proportion of calves with detectable IFN-α concentrations for the vaccine A group were significantly greater than those for the vaccine B and control groups. On day 42, the mean NF anti-P multocida IgA titers for both vaccine groups were significantly greater than that of the control group. Both vaccines induced innate and acquired immune responses in calves with colostral antibodies. The magnitude of the IFN-α response and proportion of calves with detectable IFN-α differed between the 2 vaccine groups. Both vaccines appeared to enhance the IgA response against P multocida.

Genetic variability of bovine herpesvirus type 4 (BoHV-4) field strains from Turkish cattle herds.

Bovine herpesvirus type 4 (BoHV‑4) is a common virus in the world that is detected in clinically ill or in apparently healthy cattle. This study provides a molecular characterization of BoHV‑4 strains from 24 cattle some showing respiratory and/or reproductive problems and some without any apparent clinical sign. This study also reported the growth properties of five BoHV‑4 field isolates. The 24 sampled cattle came from 13 different herds in 10 provinces collected between 2007 and 2018. Phylogenetic analysis using partially amplified nucleotide sequences of ORF8 genes coding glycoprotein B (n = 24) and ORF3 genes coding thymidine kinase (n = 9), demonstrated genetic variability among the BoHV‑4 strains analysed. The partial gB gene sequences clustered in three different genotypes (genotype I, II and III) were located within the genotype I cluster, such as Movar strain. The analysis of the five BoHV‑4 strains isolated from vaginal swabs (n = 2), nasal swab (n = 1), and brain samples (n = 2) revealed no significant differences in their growth properties in MDBK cell culture.