Bovine Herpesvirus Research Articles

Construction and Analysis of ceRNA Networks Reveal the Key Genes Associated with Bovine Herpesvirus Type 1 Infection.

Virus infection can cause the changes of lncRNA expression levels to regulate the interaction between virus and host, but the relationship between BHV-1 infection and lncRNA has not been reported. In this study, in order to reveal the molecular mechanism of RNA in BoHV-1 infection, the Madin-Darby bovine kidney (MDBK) cells were infected with BoHV-1, transcriptome sequencing were performed by next-generation sequencing at 18 h or 24 h or 33 h of viral infection and then based on the competitive endogenous RNA (ceRNA) theory, lncRNA-miRNA-mRNA networks were constructed using these high-throughput sequencing data. The network analysis, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed for functional annotation and exploration of ncRNA ceRNAs in BoHV-1 infection. The results showed that 48 lncRNAs, 123 mRNAs and 20 miRNAs as differentially expressed genes, and the mitogen activated protein kinase (MAPK) pathway and calcium signaling pathway were significantly enriched in the ceRNA network. Some differentially expressed lncRNA genes were randomly selected for verification by RT-qPCR, and the results showed that their expression trend was consistent with the results of transcriptome sequencing data. This study revealed that BoHV-1 infection can affect the expression of RNAs in MDBK cells and the regulation of ceRNA network to carry out corresponding biological functions in the host, but further experimental studies are still necessary to prove the hub genes function in ceRNA network and the molecular mechanism in BoHV-1 infection.

Characterization of Razi Bovine Kidney (RBK) Cell Line as a Sensitive Cell to Bovine Herpesvirus-1 (BoHV-1)

Viruses are obligate parasites and are completely dependent on host cells for survival and replication. RBK cell line developed and introduced by Razi Vaccine and Serum Research Institute, has been successfully established as a continuous cell line over successive passages.Viruses are obligate parasites that completely rely on host cells for survival and replication. Razi Bovine Kidney (RBK) cell line was introduced and developed by the Razi Vaccine and Serum Research Institute. It has been successfully established as a continuous cell line over successive passages. As demonstrated in this experimental study, the RBK cells have shown suitable sensitivity to certain viruses, including Bovine Herpesvirus-1 (BoHV-1) virus. In the present research, the RBK cell line characteristics were analyzed using molecular and karyotype methods and growth specifications. Cloning of the RBK cell line was performed using the limited dilution method, and each cell clone was quantitatively and qualitatively characterized. Four cell clones were compared based on their sensitivity to the BoHV-1 virus. Then, the RBK-D5 clone was selected as the most appropriate cell line for further studies. The RBK-D5 was subjected to tests for identity, chromosomal analysis, and doubling time. In the end, the origin of this cell line was confirmed by the PCR method. It was observed that the cell line exhibited karyotype diversity due to aneuploidy, which can be responsible for the procreation of chromosomal instability. This diversity represents chromosomal changes in the continuous cell line that carries the characteristic of an immortalized cell line. The RBK-D5 was found to be more sensitive to the BOHV-1 virus. Surprisingly, its titer was evaluated at 108.5 CCID50/ml. The obtained results suggested that the RBK cell line is suitable for the BoHV-1 virus and can be useful for virus detection, propagation, and quality control or viral titration.

An indirect ELISA to detect antibodies to the gC of bovine alphaherpesvirus 1 (BoAHV1) displaying no crossreactivity with antibodies induced by bovine alphaherpesvirus 5 (BoAHV5)

Seroprevalence of bovine alphaherpesvirus type 1 (BoAHV1) infections may be contaminated by crossreactive antibodies to bovine alphaherpesvirus type 5 (BoAHV5). To avoid such crossreactivity, an indirect enzyme-linked immunosorbent assay prepared with a recombinant glycoprotein C (gC) antigen (ELISA-gC1) was developed, aiming the detection of antibodies to BoAHV1, with no crossreactivity with BoAHV5 antibodies. The antigen for the ELISA-gC1 was the product of the expression of 219 bp from the N-terminal portion of the BoAHV1 gC gene, which bears low homology between the two virus types. The test was validated on 131 bovine serum samples, including 26 sera from BoAHV1-experimentally immunized, 38 sera from BoAHV5-experimentally infected or immunized calves, and 67 sera from calves seronegative for both BoAHV1 and BoAHV5, as determined by serum neutralization (SN). When compared to SN for BoAHV1, the ELISA-gC1 presented 100% sensitivity, 95.5 % specificity, 100 % negative predictive value, 89.6 % positive predictive value, 98.8 % precision, and a kappa correlation coefficient (κ) 0.95. None of the 38 BoAHV5-seropositive calves was detected by the ELISA-gC1. The ELISA-gC1 proved highly effective for the identification of BoAHV1-positive sera, with no crossreactivity with anti-BoAHV5 antibodies, thus able to distinguish serological responses from BoAHV1- and BoAHV5-seropositive cattle. Its capacity to detect BoAHV1-specific antibodies should allow the determination of the actual BoAHV1 prevalence in herds, which cannot be serologically determined in countries where BoAHV5 is also prevalent due to antibody crossreactivity. Apart from recognizing exclusively BoAHV1-infected cattle, the ELISA-gC1 may also be used in support of BoAHV5 epidemiological studies by allowing the exclusion of BoAHV1-seropositive animals.

Several Alphaherpesviruses Interact Similarly with the NF-κB Pathway and Suppress NF-κB-Dependent Gene Expression.

Alphaherpesvirus infection is associated with attenuation of different aspects of the host innate immune response that is elicited to confine primary infections at the mucosal epithelia. Here, we report that infection of epithelial cells with several alphaherpesviruses of different species, including herpes simplex virus 1 and 2 (HSV-1 and HSV-2), feline alphaherpesvirus 1 (FHV-1), and bovine alphaherpesvirus 1 (BoHV-1) results in the inactivation of the responses driven by the nuclear factor kappa B (NF-κB) pathway, considered a pillar of the innate immune response. The mode to interact with and circumvent NF-κB-driven responses in infected epithelial cells is seemingly conserved in human, feline, and porcine alphaherpesviruses, consisting of a persistent activation of the NF-κB cascade but a potent repression of NF-κB-dependent transcription activity, which relies on replication of viral genomes. However, BoHV-1 apparently deviates from the other investigated members of the taxon in this respect, as BoHV-1-infected epithelial cells do not display the persistent NF-κB activation observed for the other alphaherpesviruses. In conclusion, this study suggests that inhibition of NF-κB transcription activity is a strategy used by several alphaherpesviruses to prevent NF-κB-driven responses in infected epithelial cells. IMPORTANCE The current study provides a side-by-side comparison of the interaction of different alphaherpesviruses with NF-κB, a key and central player in the (proinflammatory) innate host response, in infected nontransformed epithelial cell lines. We report that all studied viruses prevent expression of the hallmark NF-κB-dependent gene IκB, often but not always via similar strategies, pointing to suppression of NF-κB-dependent host gene expression in infected epithelial cells as a common and therefore likely important aspect of alphaherpesviruses.

Bovine myeloid antimicrobial peptide-28 (BMAP-28) mRNA expression by bovine cells and effects of synthetic MAP-28 on bovine respiratory disease pathogens

Mannheimia haemolytica (MH) is the principal bacterial patho­gen associated with bovine respiratory disease (BRD) in beef cattle. Bovine Herpes virus type 1 (BHV-1) can cause BRD by itself or it can predispose cattle to BRD due to MH or other op­portunistic bacteria. Existing antimicrobials do not consis­tently prevent BRD due to MH, and do not have an effect against viruses; bovine antimicrobial peptides (AMP) have immune stimulating and nonspecific antimicrobial effects that could improve BRD control. Messenger RNA (mRNA) treatment could be used to induce AMP expression in cattle, but efficacy must first be confirmed in vitro. Synthetic AMP can be generated to use as standards when characterizing mRNA-expressed AMP. We hypothesized that bovine cells can express synthetic mRNA coding for the AMP BMAP-28 and that synthetic BMAP-28 can inhibit the growth of MH and elicit antiviral effects against BHV-1 virus.

Seroepidemiological study of bovine alphaherpesvirus 1 in the dairy cattle herds of Addis Ababa, Ethiopia.

Bovine alphaherpesvirus 1(BoHV-1) causes respiratory disease, abortions, and genital disorders in cattle. Although BoHV-1 has been known to cause severe economic damage to the dairy industries, little is known about its epidemiology in dairy cattle of Addis Ababa, Ethiopia. The present study aimed to determine the seroprevalence and the risk factors associated with the occurrence of BoHV-1. A total of 369 blood samples from 115 dairy herds were collected using a proportional stratified random sampling method and examined antibodies against BoHV-1 using ELISA test. A questionnaire survey was done to gather information related to farm demographics and reproductive disorders. Univariate and multivariate mixed-effect logistic regression analyses were used. The overall seroprevalence of BoHV-1 was detected in 21 % (95%CI: 17-25%) and 32 % (95%CI: 24-42 %) at animal and herd levels, respectively. A multivariable mixed effect logistic regression model revealed that adult cattle had 14 times (OR = 14.32; 95 % CI: 2.53-81.5; P=0.003) more likely to increase the risk of being BoHV-1 seropositive than young cattle. Purchased cattle had 4 times (OR = 4.15; 95 % CI: 1.36-12.66, P=0.012) more likely to increase the risk of being BoHV-1 seropositive than homebred cattle. The risk of being BoHV-1 seropositive was 195 times higher in herds using bulls (OR = 195.51; 95 % CI: 3.62-1056.51; P=0.010) than in herds using artificial insemination only for breeding. BoHV-1 seropositivity was significantly associated with cows that had a history of abortion (OR = 6.89; 95 % CI: 1.97-22.76; P=0.002), retained placenta (OR = 3.26; 95 % CI: 1.32-8.07; P=0.010), and repeat breeding (OR = 3.64; 95 % CI: 1.08-12.18; P=0.036). This study demonstrated the gaps in the selection of BoHV-1 free bulls for breeding as well as limited farm biosecurity practices. Thus, awareness creation for dairy farmers on good farm biosecurity practices including vaccination should be initiated.

BoHV-1 affects abortion and progesterone in dairy cows Bovine alphaherpesvirus 1 (BoHV-1) seropositivity, progesterone levels and embryo loss of 30-day-old pregnant dairy cows in Zagros Industrial Dairy Farm in Shahrekord: Examination and analysis.

Bovine alphaherpesvirus 1 (BoHV-1) is a serious disease with severe negative economic effects on the global cattle sector, especially in Iran. This cross-sectional study was carried out to examine the seroprevalence and associated risk factors of BoHV-1 infection with progesterone levels and embryo death in 30-day pregnant dairy cattle at Zagros Industrial Dairy Farm in Shahrekord, Iran. Between December 2017 to February 2018, blood samples were obtained from 60 dairy cow herds. To determine whether BoHV-1 was present, serum samples were examined using the ELISA for serum antibodies. To find progesterone (P4) in blood, the progesterone ELISA test was used. 96.7 % of sera tested positive for BoHV-1 antibodies, according to the findings. Additionally, 60.34 % of blood samples that tested positive had an experience of abortion and significantly more inseminations that resulted in pregnancy, consistent with findings from other studies conducted in Iran and other nations. Since this study is the first to document the risk factor for BoHV-1 infection in Shahrekord, Iran, we could infer that the virus is extensively dispersed in this area.

A Rapid Nucleic Acid Visualization Assay for Infectious Bovine Rhinotracheitis Virus That Targets the TK Gene.

Infectious bovine rhinotracheitis virus (IBRV) can cause various degrees of symptoms in the respiratory system, reproductive system, and whole body of cattle. It also can lead to persistent and latent infection in cattle, posing a challenge to timely control of infectious bovine rhinotracheitis (IBR) in farms and causing large financial losses in the global cattle industry. Therefore, the goal of this study was to establish a rapid, simple, and accurate method that can detect IBRV in order to facilitate the control and eradication of IBR in cattle. We combined recombinant polymerase amplification (RPA) with a closed vertical flow visualization strip (VF) and established an RPA-VF assay that targets the thymidine kinase (TK) gene to rapidly detect IBRV. This method (reaction at 42°C for 25 min) was able to detect a minimum of 3.8 × 101 copies/μL of positive plasmid and 1.09 × 101 50% tissue culture infective dose (TCID50) of the IBRV. This assay has high specificity for IBRV and does not cross-react with other respiratory pathogens in cattle. The concordance between the RPA-VF assay and the gold standard was 100%. In addition, this assay was also suitable for the detection of DNA from clinical samples extracted by a simple method (heating at 95°C for 5 min), which can achieve the rapid detection of clinical samples in the field. Overall, the present sensitivity, specificity, and clinical applicability assessments indicated that the RPA-VF assay we developed can be utilized as a quick and accurate on-site test for IBRV detection in farms. IMPORTANCE IBRV causes different degrees of clinical symptoms in cattle and poses a great threat to the cattle industry. The infection is persistent and latent, and the elimination of IBRV in infected herds is difficult. A rapid, simple, and accurate method to detect IBRV is therefore vital to control and eradicate IBR. Combining RPA with an VF, we established an RPA-VF assay for the rapid detection of IBRV, which can complete the test of clinical samples in 35 min. The assay shows good sensitivity, specificity, and clinical applicability and can be used as an on-site test for IBRV in farms.

Humoral Immune Response of Mice against a Vaccine Candidate Composed of a Chimera of gB of Bovine Alphaherpesviruses 1 and 5.

Infectious bovine rhinotracheitis (IBR) and bovine meningoencephalitis are caused by Bovine alphaherpesvirus (BoHV) types 1 and 5, which seriously threaten the global cattle industry. Vaccination to improve immunity is the most direct and effective means to prevent these conditions. Glycoprotein B (gB) is essential for the attachment of both viruses to permissive cells, and is a major target of the host immune system, inducing a strong humoral response. The aim of this study was to evaluate, in a murine model, the immune response of a candidate vaccine formulation composed of a chimeric BoHV-1 and BoHV-5 gB (DgB), expressed in Komagataella phaffii. The chimeric DgB vaccine adjuvanted with Montanide 50 ISA V2 or aluminum hydroxide was administered intramuscularly or subcutaneously. A control group and a group that received a commercial vaccine were inoculated subcutaneously. Higher titers of neutralizing antibodies against BoHV-1, BoHV-5, and a natural BoHV-1/5 recombinant strain were obtained with the oil-based candidate vaccine formulation administered intramuscularly. The results demonstrated that the chimeric DgB conserved important epitopes that were able to stimulate a humoral immune response capable of neutralizing BoHV-1, BoHV-5, and the recombinant strain, suggesting that the vaccine antigen is a promising candidate to be further evaluated in cattle.

Construction of a one-step multiplex real-time PCR assay for the detection of serogroups A, B, and E of Pasteurella multocida associated with bovine pasteurellosis.

Bovine pasteurellosis, caused by serogroups A, B, and E of Pasteurella multocida (Pm), is mainly manifested as bovine respiratory disease (BRD) and hemorrhagic septicemia (HS). The disease has caused a great economic loss for the cattle industry globally. Therefore, identifying the Pm serogroups is critical for optimal diagnosis and subsequent clinical treatment and even epidemiological studies. In this study, a one-step multiplex real-time PCR assay was established. Three pairs of specific primers were prepared to detect the highly conserved genomic regions of serogroups A (HyaD), B (bcbD), and E (ecbJ) of Pm, respectively. The results depicted that the method had no cross-reaction with other bovine pathogens (Mannheimia hemolytica, Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, Salmonella Dublin, Mycobacterium paratuberculosis, infectious bovine rhinotracheitis virus, and Mycoplasma bovis). The linear range (107 to 102 copies/μL) showed the R2 values for serogroups A, B, and E of Pm as 0.9975, 0.9964, and 0.996, respectively. The multiplex real-time PCR efficiency was 90.30%, 90.72%, and 90.57% for CartA, CartB, and CartE, respectively. The sensitivity result showed that the serogroups A, B, and E of Pm could be detected to be as low as 10 copies/μL. The repeatability result clarified that an intra-assay and an inter-assay coefficient of variation of serogroups A, B, and E of Pm was < 2%. For the clinical samples, the detection rate was higher than the OIE-recommended ordinary PCR. Overall, the established one-step multiplex real-time PCR assay may be a valuable tool for the rapid and early detection of the serogroups A, B, and E of Pm with high specificity and sensitivity.

A novel triplex real-time PCR assay for the differentiation of lumpy skin disease virus, goatpox virus, and sheeppox virus.

Three members of Capripoxvirus (CaPV) genus, including lumpy skin disease virus (LSDV), goatpox virus (GTPV), and sheeppox virus (SPPV), are mentioned as notifiable forms by World Organization for Animal Health. These viruses have negatively impacted ruminant farming industry worldwide, causing great economic losses. Although SPPV and GTPV cause more severe clinical disease in only one animal species, they can transfer between sheep and goats. Both homologous and heterologous immunization strategies are used to protect animals against CaPVs. However, development of accurate and rapid methods to distinguish these three viruses is helpful for the early detection, disease surveillance, and control of CaPV infection. Therefore, we developed a novel triplex real-time PCR (qPCR) for the differentiation of LSDV, GTPV, and SPPV. Universal primers were designed to detect pan-CaPV sequences. Species-specific minor groove binder (MGB)-based probes were designed, which were labeled with FAM for LSDV, HEX for GTPV, and ROX for SPPV. The sensitivity, specificity, reproducibility, and ability of detecting mixed infections were evaluated for the triplex qPCR. Further, 226 clinical samples of the infection and negative controls were subjected to the triplex qPCR, and the results were verified using PCR-restriction fragment length polymorphism (PCR-RFLP) and sequencing methods for PRO30 gene. The triplex qPCR could successfully distinguish LSDV, GTPV, and SPPV in one reaction, and the assay sensitivity was 5.41, 27.70, and 17.28 copies/μL, respectively. No cross-reactivity was observed with other viruses causing common ruminant diseases, including des petits ruminants virus, foot-and-mouth disease virus, bluetongue virus, ovine contagious pustular dermatitis virus, infectious bovine rhinotracheitis virus, and bovine viral diarrhea-mucosal disease virus. Inter-and intra-assay variabilities were < 2.5%. The results indicated that the triplex qPCR was highly specific, sensitive, and reproducible. Simulation experiments revealed that this assay could successfully distinguish two or three viruses in case of mixed infections without any cross-reaction. For clinical samples, the results were completely consistent with the results of PCR-RFLP and sequencing. This demonstrated that the assay was reliable for clinical application. The triplex qPCR is a robust, rapid, and simple tool for identifying various types of CaPV as it can successfully distinguish LSDV, GTPV, and SPPV in one reaction. Furthermore, the assay can facilitate more accurate disease diagnosis and surveillance for better control of CaPV infection.

The bovine nasal fungal community and associations with bovine respiratory disease.

Effective identification and treatment of bovine respiratory disease (BRD) is an ongoing health and economic issue for the dairy and beef cattle industries. Bacteria pathogens Pasteurellamultocida, Mycoplasmabovis, Mannheimia haemolytica, and Histophilus somni and the virus Bovine herpesvirus-1 (BHV-1), Bovine parainfluenza-3 virus (BPIV-3), Bovine respiratory syncytial virus (BRSV), Bovine adenovirus 3 (BAdV3), bovine coronavirus (BoCV) and Bovine viral diarrhea virus (BVDV) have commonly been identified in BRD cattle; however, no studies have investigated the fungal community and how it may also relate to BRD. The objective of this study was to understand if the nasal mycobiome differs between a BRD-affected (n = 56) and visually healthy (n = 73) Holstein steers. Fungal nasal community was determined by using Internal Transcribed Spacer (ITS) sequencing. The phyla, Ascomycota and Basidiomycota, and the genera, Trichosporon and Issatchenkia, were the most abundant among all animals, regardless of health status. We identified differences between healthy and BRD animals in abundance of Trichosporon and Issatchenkia orientalis at a sub-species level that could be a potential indicator of BRD. No differences were observed in the nasal fungal alpha and beta diversity between BRD and healthy animals. However, the fungal community structure was affected based on season, specifically when comparing samples collected in the summer to the winter season. We then performed a random forest model, based on the fungal community and abundance of the BRD-pathobionts (qPCR data generated from a previous study using the same animals), to classify healthy and BRD animals and determine the agreement with visual diagnosis. Classification of BRD or healthy animals using ITS sequencing was low and agreed with the visual diagnosis with an accuracy of 51.9%. A portion of the ITS-predicted BRD animals were not predicted based on the abundance of BRD pathobionts. Lastly, fungal and bacterial co-occurrence were more common in BRD animals than healthy animals. The results from this novel study provide a baseline understanding of the fungal diversity and composition in the nasal cavity of BRD and healthy animals, upon which future interaction studies, including other nasal microbiome members to further understand and accurately diagnose BRD, can be designed.

Investigation of the Role of Some Selected Virus Species in Abnormality of Calves

Early embryonic death, fetal resorption, fetal mummification/maceration, abortion, and abnormalities are some of the causes of losses in farm animals. In this study, it was aimed to make recommendations to clinician veterinarians by investigating the presence of bovine viral diarrhea virus (BVDV), Schmallenberg virus (SBV), Akabane virus (AKAV), and bovine herpesvirus-1 (BHV-1) viruses in calves with congenital anomalies. A total of 34 calves of different breeds and sexes, between 1 day and 2 months of age which were brought to Fırat University Animal Hospital for diagnosis and treatment and found to have congenital anomalies after a general clinical examination during visits to the neighboring villages, were included in the study. The abnormalities detected in the calves were as follows: Hydrocephalus, cerebellar hypoplasia, brahignati inferior, mandibular laterognathism, arthrogryposis, cheilognathopalatoschisis, coccygeal agenesis, atresia ani, bouleture/arqure, ocular and orbital deformities, dermoid cysts, miniature, renal and urinary system anomalies. PCR results showed pestivirus was detected in 6 of 34 (17.6%) calves with abnormalities. After sequencing, the expected PCR product (288 nucleotides) for pestivirus showed high identity (97% and above) to the BVDV-1 strain/isolates in the GenBank. In conclusion, although the primary purpose of cattle breeding is high meat and milk yield, it is also very important to continue the production process with healthy newborn calves. For this reason, even if congenital anomalies are seen at a low rate in calves, they should not be underestimated as they will cause economic losses, and we believe that clinician veterinarians should not ignore these diseases.

Nosological profile and classification of infectious diseases of farm animals in the territory of the Pepublic of Sakha (Yakutia)

The results of the classification of infectious diseases of farm animals spread on the territory of the Republic of Sakha (Yakutia) for the period 2001-2021 are presented. It was found that there were 42 nosological units of animal diseases involved in the epizootic process on the territory of Yakutia. Infections in cattle are caused by 18 types of pathogens, horses - 12, reindeer - 4, pigs - 10, birds - 11, small ruminants - 3, fur-bearing animals - 7, other types of animals - 9. According to the OIE classification, 10 nosoforms of diseases are common in Yakutia: rabies, brucellosis, viral hemorrhagic septicemia, equine infectious anemia, avian infectious bronchitis, infectious bovine rhinotracheitis, paratuberculosis, pasteurellosis and tuberculosis. Diseases not on the OIE list, including Marek’s disease, swine dysentery, colibacillosis, parvoviral enteritis, salmonellosis, diamond-skin disease, blackleg were registered. A number of nosoforms common in Yakutia belong to particularly dangerous diseases for which restrictive measures (quarantine) may be imposed. These include actinomycosis, rabies, brucellosis, infectious equine anemia, avian infectious bronchitis, infectious bovine rhinotracheitis, bovine leukosis, leptospirosis, paratuberculosis, diamond-skin disease, tuberculosis and canine distemper virus. A number of diseases (Marek’s disease, carnivorous infectious hepatitis, bovine leukosis, horse strangles, avian ornithosis, egg drop syndrome, diamond-skin disease, and blackleg) are peculiar to only one species of animals. Many infections are characterized by the persistence of the pathogen in different species of farm and small domestic animals, birds and wildlife, which determines the interspecies degree of epizootic danger of these infections. Infections of bacterial etiology (bacterioses) prevail in Yakutia with 21 nosoforms. Viral diseases (viroses) are represented by 15 nosoforms, mycoplasmosis and chlamydia - one each. The most numerous infections are those with the main alimentary, or fecal-oral, route (22 diseases). Ten infections, including infectious atrophic rhinitis, infectious bovine rhinotracheitis, distemper, are classified as airborne or respiratory (major) pathogen transmission. Transmissive carry-over (by vectors) is characterized by actinomycosis and infectious equine anemia; through the outer covers (without the participation of vectors) - 5 diseases, including balanoposthitis and rabies. The majority of the infections are characterized by one or more additional mechanisms of transmission (rabies, Marek’s disease, bovine leukemia, diamond-skin disease, etc.) in addition to the main mechanism of transmission of the pathogen. According to the source of the causative agent for all diseases (ktenonoses) they are domestic animals, but for such infections as rabies, Marek’s disease, viral hemorrhagic septicemia, leptospirosis and others, along with domestic animals, wild animals are an additional source (ktenotherionoses). In terms of susceptibility, most infections (30) belong to zoonoses peculiar only to animals; the remaining 12 belong to the more dangerous zooanthroponoses common to animals and humans.

Fosfomycin modifies the replication kinetics of bovine alphaherpesvirus-1 and reduces the timing of its protein expression on bovine (MDBK) and human (SH-SY5Y) cell lines.

Bovine alphaherpesvirus 1 (BoAHV-1) predisposes cattle to respiratory secondary bacterial infections, which can be treated with the broad-spectrum antibiotic fosfomycin. This drug also suppresses NF-kB activity and pro-inflammatory responses. Therefore, cattle may be exposed to an interaction between the virus and the antibiotic which may have effects on it. The aim of this study was to determine the effect of calcium fosfomycin (580µg/mL) on BoAHV-1 (moi = 0.1) replication. Two cell lines (MDBK and SH-SY5Y) were used in this study. Our results show that fosfomycin has novel properties. By MTT assay we have shown that it is non-cytotoxic for any of the cell lines. Extracellular and intracellular viral titers demonstrated that fosfomycin has a cell-type and time-dependent effect on BoAHV-1 replication. By direct immunofluorescence it was shown that it reduces the timing of BoAHV-1 protein expression, and by qPCR, we found that its effect on NF-kB mRNA expression depends on the cell type.

Preliminary Evaluation of Lablab purpureus Phytochemicals for Anti-BoHV-1 Activity Using In Vitro and In Silico Approaches.

Lablab purpureus from the Fabaceae family has been reported to have antiviral properties and used in traditional medical systems like ayurveda and Chinese medicine and has been employed to treat a variety of illnesses including cholera, food poisoning, diarrhea, and phlegmatic diseases. The bovine alphaherpesvirus-1 (BoHV-1) is notorious for causing significant harm to the veterinary and agriculture industries. The removal of the contagious BoHV-1 from host organs, particularly in those reservoir creatures, has required the use of antiviral drugs that target infected cells. This study developed LP-CuO NPs from methanolic crude extracts, and FTIR, SEM, and EDX analyses were used to confirm their formation. SEM analysis revealed that the LP-CuO NPs had a spherical shape with particle sizes between 22 and 30 nm. Energy-dispersive X-ray pattern analysis revealed the presence of only copper and oxide ions. By preventing viral cytopathic effects in the Madin-Darby bovine kidney cell line, the methanolic extract of Lablab purpureus and LP-CuO NPs demonstrated a remarkable dose-dependent anti-BoHV-1 action in vitro. Furthermore, molecular docking and molecular dynamics simulation studies of bio-actives from Lablab purpureus against the BoHV-1 viral envelope glycoprotein disclosed effective interactions between all phytochemicals and the protein, although kievitone was found to have the highest binding affinity, with the greatest number of interactions, which was also validated with molecular dynamics simulation studies. Understanding the chemical reactivity qualities of the four ligands was taken into consideration facilitated by the global and local descriptors, which aimed to predict the chemical reactivity descriptors of the studied molecules through the conceptual DFT methodology, which, along with ADMET finding, support the in vitro and in silico results.

A scoping review of the testing of bulk tank milk to detect nonbacterial pathogens or herd exposure to nonbacterial pathogens in dairy cattle

In this scoping review, we characterized the literature reporting on the testing of bulk milk samples to detect microorganisms other than bacteria that can cause diseases in dairy cattle, including viruses, helminths, algae, and protozoa. A search strategy was completed by screening databases, conference proceedings, animal health agency websites, disease surveillance program websites, and handbooks of cattle-related diagnostic tests for potentially relevant articles. Two reviewers independently screened articles in English, Portuguese, or Spanish; original studies reporting on the testing of farm-level, unprocessed bulk milk samples for presence of pathogens or specific antibodies against agents other than bacteria that can cause diseases in cows were retained. From all studies, we used spreadsheets to extract relevant information, including pathogen screened, test used, and country of origin of bulk milk samples. Additionally, for studies reporting sufficient data to estimate test characteristics, we extracted detailed information about herd eligibility, testing protocol, and herd-level infection definition. A total of 8,829 records were identified, from which 1,592 were retained and assessed for eligibility, and 306 were included. Bovine viral diarrhea virus, Fasciola hepatica, Ostertagia ostertagi, and bovine herpesvirus 1 were the most frequently screened agents, reported from 107, 45, 45, and 33 studies, respectively. Sensitivity of bulk milk ELISA to detect herds with animals infected by bovine herpesvirus 1 ranged from 2 to 100%, and was affected mostly by antigen selection, cut-off adopted, herd vaccination status, and seroprevalence of lactating cows. Bulk milk ELISA had very high specificity to detect herds free of bovine leukemia virus, and varying sensitivity to detect herds with infected animals, which depended on the within-herd seroprevalence of lactating cattle. As for bovine viral diarrhea virus, in general, the sensitivity of bulk milk ELISA was moderate to high (>80%) when infection status was defined based on presence of persistently infected cattle or a high proportion of seropositive lactating cattle. Nevertheless, bulk milk ELISA was not able to distinguish infected and noninfected herds based on presence of seropositive unvaccinated weanlings. The PCR or quantitative PCR protocols employed had very low sensitivities (<40%) and very high specificities (>95%) to classify bovine viral diarrhea virus infection status of dairy herds. Sensitivity and specificity of bulk milk ELISA to classify herds with regards to presence of F. hepatica- or O. ostertagi-parasitized cattle were generally high and driven mostly by the definition of herd infection status. Conversely, bulk milk ELISA demonstrated varying characteristics to detect herds with or without Dictyocaulus viviparus-parasitized cattle, depending primarily on the antigen selected and presence of cattle with clinical signs of lungworm infection.

Endogenous and viral microRNAs in nasal secretions of water buffaloes (Bubalusbubalis) after Bubalinealphaherpesvirus1 (BuHV-1) challenge infection

Bubaline alphaherpesvirus 1 (BuHV-1) is a pathogen of water buffaloes responsible for economic loss worldwide. MicroRNAs (miRNAs) regulate gene expression produced by alphaherpesviruses and hosts. This study aimed at (a) unravelling the ability of BuHV-1 to produce miRNAs, including hv1-miR-B6, hv1-miR-B8, hv1-miR-B9; (b) measuring the host immune-related miRNAs associated to herpesvirus infection, including miR-210-3p, miR-490-3p, miR-17-5p, miR-148a-3p, miR-338-3p, miR-370-3p, by RT-qPCR; (c) identifying candidate markers of infection by receiver-operating characteristic (ROC) curves; (d) exploiting the biological functions by pathway enrichment analyses. Five water buffaloes BuHV-1 and Bovine alphaherpesvirus 1 (BoHV-1) free were immunized against Infectious Bovine Rhinotracheitis (IBR). Five additional water buffaloes served as negative controls. All animals were challenged with a virulent wild-type (wt) BuHV-1 via the intranasal route 120 days after the first vaccination. Nasal swabs were obtained at days (d) 0, 2, 4, 7, 10, 15, 30, and 63 post-challenge (pc). The animals of both groups shed wt BuHV-1 up to d7 pc. Results demonstrated that (a) miRNAs produced by the host and BuHV-1 could be efficiently quantified in the nasal secretion up to d63 and d15 pc, respectively; b) the levels of host and BuHV-1 miRNAs are different between vaccinated and control buffaloes; c) miR-370-3p discriminated vaccinated and control animals; d) host immune-related miRNAs may modulate genes involved in the cell adhesion pathway of the neuronal and immune system. Overall, the present study provides evidence that miRNAs can be detected in nasal secretions of water buffaloes and that their expression is modulated by BuHV-1.

A serological survey of pathogens associated with the respiratory and digestive system in the Polish European bison (Bison bonasus) population in 2017–2022

BackgroundThe European bison (Bison bonasus) is a near threatened species and requires health monitoring. The aim of the present study was to determine the prevalence of antibodies to pathogens known to cause respiratory and digestive illness in ruminants.ResultsIn the studied 328 European bison, the highest seroprevalence was observed for Bovine herpesvirus-1 (BoHV-1) (50.27%), Bovine Coronavirus (BCoV) (26.36%), and Bluetongue Virus (BTV) (12.83%). For Mycoplasma bovis strains and Bovine Viral Diarrhea Virus (BVDV), positive results were rare. Interestingly, a higher prevalence of BTV antibodies was noted in the northeastern populations and older animals.ConclusionsOur findings indicate that the Polish European bison population appears to have considerable contact with BoHV-1; however, this does not appear to be of great significance, as clinical symptoms and post-mortem lesions are rarely noted in Polish European bison population. The high seroprevalence of BTV in the north-east of Poland is an ongoing trend, also noted in previous studies. It is possible that European bison may perpetuate the virus in this region. This is the first report of antibodies for BCoV in European bison.

Bovine Airway Models: Approaches for Investigating Bovine Respiratory Disease.

Bovine respiratory disease (BRD) is a multifactorial condition where different genera of bacteria, such as Mannheimia haemolytica, Histophilus somni, Pasteurella multocida, and Mycoplasma bovis, and viruses, like bovine respiratory syncytial virus, bovine viral diarrhea virus, and bovine herpes virus-1, infect the lower respiratory tract of cattle. These pathogens can co-infect cells in the respiratory system, thereby making specific treatment very difficult. Currently, the most common models for studying BRD include a submerged tissue culture (STC), where monolayers of epithelial cells are typically covered either in cellular or spent biofilm culture medium. Another model is an air-liquid interface (ALI), where epithelial cells are exposed on their apical side and allowed to differentiate. However, limited work has been reported on the study of three-dimensional (3D) bovine models that incorporate multiple cell types to represent the architecture of the respiratory tract. The roles of different defense mechanisms in an infected bovine respiratory system, such as mucin production, tight junction barriers, and the production of antimicrobial peptides in in vitro cultures require further investigation in order to provide a comprehensive understanding of the disease pathogenesis. In this report, we describe the different aspects of BRD, including the most implicated pathogens and the respiratory tract, which are important to incorporate in disease models assembled in vitro. Although current advancements of bovine respiratory cultures have led to knowledge of the disease, 3D multicellular organoids that better recapitulate the in vivo environment exhibit potential for future investigations.