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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.

Aims: The present study aimed to Searching Bovine Herpesvirus Types 1, 4 and Bovine Parainfluenza Virus Type 3 Infections in Milk of Cows with Clinical Mastitis Based in Dairy Cattle Managements. Study Design: In 35 managements around Burdur region, milk of 123 cows with clinical mastitis was searched for Bovine Herpesvirus Types 1 (BHV-1), 4 (BHV-4) and Bovine Parainfluenza Virus Type 3 (BPIV-3) infections. Results: In the study, the highest seropositivity was detected against BPIV-3. The highest seropositivity on infection distribution according to age was found against every four viruses in animals within the three-year-old group. The highest seropositivity in this group and other age groups was detected against BPIV-3. Seropositivity against these viruses was found in the highest right anterior one and the lowest left posterior one out of determined udder lobes. The highest seropositivity was found in semi-outdoor managements with concrete and dirty grounds where cleaning/disinfection of teats before and after milking was performed, mastitis treatment and viral vaccination was not applied, the ground of stable was cleaned monthly, only feces was collected from the grounds, water was used for cleaning of milking machines and hands and iodised disinfectant was used. During udder inspection evaluation of animals with clinical mastitis, the highest seropositivity was detected in the ones having normal udder, teat and skin appearance. Out of udder and teat skin lesions, the highest seropositivity was found in crushed ones. In the evaluation of udder palpation in animals with clinical mastitis, the highest seropositivity was detected in the ones with teat tissue thickening and with elastic teat sinuses and lobes. In the milk of these animals, the highest seropositivity was found in the ones showing coagulation. Conclusion: It was stated that viruses took a vital part in clinical mastitis cases, the structure, practice, cleaning and disinfection of managements was really important, udder and teat skin lesions, tissue thickening, elastic teat sinuses and lobes and coagulation of milk was also crucial. Besides, the appearance of udder, teat and skin was not relevant.

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.

Background Bovine herpesvirus type 5 is an important agent of meningoencephalitis in cattle and has been identified in outbreaks of bovine neurological disease in several Brazilian states. In recent years, oxoquinoline derivatives have become an important focus in antiviral drug research. Methods The cytotoxicity and anti BoHV-5RJ42/01 activity of a set of synthetic 4-oxoquinoline derivatives 4a-k were assayed on Madin–Darby Bovine Kidney cell and antiviral activity by plaque reduction assay. Results: The most promising substance (4h) exhibited CC50 and EC50 values of 1,239 μM ±5.5 and 6.0 μM ±1.5, respectively, with an SI =206. Two other compounds 4j (CC50 = 35 μM ±2 and EC50 = 24 μM ±7.0) and 4k (CC50= 55 μM ±2 and EC50 = 24 μM ±5.1) presented similar inhibitory profile and selectivity indexes of 1.4 and 2.9, respectively. The results of the time-of-addition studies revealed expressive reduction of virus production (≥80%) in different stages of virus replication cycle except for compound 4h that slightly inhibited virus yield in the first 2 h post infection, but it showed expressive virus inhibition after this time. Conclusions All three compounds slightly interact with the virus on the virucidal assay and they are not able to block virus attachment and penetration. Antiviral effect of oxoquinoline 4h was more prominent than acyclovir which leads us to suggest compound 4h as a promising molecule for further anti-BoHV-5 drug design.

En México no se han realizado estudios serológicos sobre herpesvirus caprino tipo 1 con el propósito de saber el estado de la infección por CpHV-1. Se llevó a cabo un análisis serológico para determinar la presencia de anticuerpos contra el virus usando un ELISA de bloqueo comercial para detección de anticuerpos contra la glicoproteína B (gB) y glicoproteína E (gE) del herpesvirus bovino tipo 1, de acuerdo con la prueba diagnóstica que se utiliza de forma rutinaria. De un total de 838 animales analizados, 123 (14.68 %) resultaron positivos a la prueba de ELISA. En los estados de Puebla, Morelos, Nuevo León, Ciudad de México, Guanajuato y Querétaro se encontró evidencia serológica de la presencia del CpHV-1 en cabras. Este reporte es el primer estudio que señala la presencia de anticuerpos contra herpesvirus caprino en cabras en México.

Infectious bovine keratoconjunctivitis (IBK), also known as pinkeye, is one of the most common eye diseases in cattle. Several pathogens have been associated with IBK cases, however, Moraxella bovis, Moraxella bovoculi, Mycoplasma bovis, Mycoplasma bovoculi and bovine herpesvirus type 1 (BHV-1) are most frequently observed. A multiplex real-time PCR assay using two reactions was developed for the detection and differentiation of these five pathogens. Detection sensitivities of the multiplex assays were compared to singleplex reactions testing for the same targets. Correlation coefficients (R2) of >0.99, and PCR efficiencies between 92 and 106% were demonstrated in all singleplex and multiplex real-time PCR reactions. The limits of detection (LOD) of multiplex assays for Moraxella bovis, Moraxella bovoculi, Mycoplasma bovis, Mycoplasma bovoculi and BHV-1 were 19, 23, 25, 24 and 26 copies per reaction, respectively. No cross amplification was observed for specificity testing of 179 IBK positive clinical samples and 55 non-target clinical samples. Percentage of clinical samples positive for Mycoplasma bovoculi, Moraxella bovoculi, Moraxella bovis, BHV-1 and Mycoplasma bovis were 88.8% (159/179), 75.9% (136/179), 60.3% (108/179), 11.7% (21/179) and 10.0% (18/179), respectively. Moraxella bovis, Moraxella bovoculi and Mycoplasma bovoculi were more prevalent than Mycoplasma bovis and BHV-1 in IBK samples collected from animals in this study population. Our data indicates that the multiplex real-time PCR panel assay is highly sensitive and highly specific for the detection and differentiation of the five major pathogens associated with bovine pinkeye., Highlights • A multiplex real-time PCR is developed for the detection of five major IBK pathogens. • Correlation coefficients of all standard curves were >0.99. • PCR amplification efficiencies for the five pathogens were between 92% and 106%. • Limits of detection were between 19 and 26 copies per reaction for the five pathogens. • Moraxella bovoculi, Mycoplasma bovoculi, and Moraxella bovis were more prevalent IBK pathogens.

Bovine Respiratory Disease (BRD) is a multifactorial condition affecting cattle worldwide resulting in high rates of morbidity and mortality. The disease can be triggered by Bovine Herpesvirus-1 (BoHV-1) infection, stress, and the subsequent proliferation and lung colonization by commensal bacteria such as Mannheimia haemolytica, ultimately inducing severe pneumonic inflammation. Due to its polymicrobial nature, the study of BRD microbes requires co-infection models. While several past studies have mostly focused on the effects of co-infection on host gene expression, we focused on the relationship between BRD pathogens during co-infection, specifically on M. haemolytica’s effect on BoHV-1 replication. This study shows that M. haemolytica negatively impacts BoHV-1 replication in a dose-dependent manner in different in vitro models. The negative effect was observed at very low bacterial doses while increasing the viral dose counteracted this effect. Viral suppression was also dependent on the time at which each microbe was introduced to the cell culture. While acidification of the culture medium did not grossly affect cell viability, it significantly inhibited viral replication. We conclude that M. haemolytica and BoHV-1 interaction is dose and time-sensitive, wherein M. haemolytica proliferation induces significant viral suppression when the viral replication program is not fully established.

International audience; Herpesvirus entry and spread requires fusion of viral and host cell membranes, which is mediated by the conserved surface glycoprotein B (gB). Upon activation, gB undergoes a major conformational change and transits from a metastable prefusion to a stable postfusion conformation. Although gB is a structural homolog of low-pH-triggered class III fusogens, its fusion activity depends strictly on the presence of the conserved regulatory gH/gL complex and nonconserved receptor binding proteins, which ensure that fusion occurs at the right time and space. How gB maintains its prefusion conformation and how gB fusogenicity is controlled remain poorly understood. Here, we report the isolation and characterization of a naturally selected pseudorabies virus (PrV) gB able to mediate efficient gH/gL-independent virus-cell and cell-cell fusion. We found that the control exerted on gB by the accompanying viral proteins is mediated via its cytosolic domain (CTD). Whereas gB variants lacking the CTD are inactive, a single mutation of a conserved asparagine residue in an alpha-helical motif of the ectodomain recently shown to be at the core of the gB prefusion trimer compensated for CTD absence and uncoupled gB from regulatory viral proteins, resulting in a hyperfusion phenotype. This phenotype was transferred to gB homologs from different alphaherpesvirus genera. Overall, our data propose a model in which the central helix acts as a molecular switch for the gB pre-to-postfusion transition by conveying the structural status of the endo-to the ectodomain, thereby governing their cross talk for fusion activation, providing a new paradigm for herpesvirus fusion regulation. IMPORTANCE The class III fusion protein glycoprotein B (gB) drives membrane fusion during entry and spread of herpesviruses. To mediate fusion, gB requires activation by the conserved gH/gL complex by a poorly defined mechanism. A detailed molecular-level understanding of herpesvirus membrane fusion is of fundamental virological interest and has considerable potential for the development of new therapeutics blocking herpesvirus cell invasion and spread. Using in vitro evolution and targeted mutagenesis of three different animal alphaherpesviruses, we identified a single conserved amino acid in a regulatory helix in the center of the gB ectodomain that enables efficient gH/gL-independent entry and plays a crucial role in the pre-to-postfusion transition of gB. Our results propose that the central helix is a key regulatory element involved in the intrastructural signal transduction between the endo- and ectodomain for fusion activation. This study expands our understanding of herpesvirus membrane fusion and uncovers potential targets for therapeutic interventions.

Background Biosecurity is a key measure to reduce and prevent the introduction of diseases to farms and minimise spread of diseases within a herd. The aim of the study was to characterise the current application of biosecurity measures on dairy cattle farms in Spain along with their bovine viral diarrhoea and infectious bovine rhinotracheitis status. Methods Data on biosecurity measures for 124 dairy herds were collected using a questionnaire. The sanitary status of these farms for bovine viral diarrhoea and infectious bovine rhinotracheitis was also assessed using antibody ELISA. Data were analysed using multiple correspondence analysis and a two-step cluster analysis. Results Three main clusters of farms were identified: clusters 1 and 2 included herds of small and intermediate sizes. These, particularly cluster 1, showed the most deficiencies in the control of vehicles and visitors. However, laboratory tests were always performed on purchased animals. Cluster 3 had the largest herd sizes, with somewhat better biosecurity control of vehicles and visitors. However, farms in this cluster also purchased the most animals, sometimes without testing, and hired external workers more often. Conclusion The study indicated that, in the study population, there are serious shortcomings in the application of biosecurity measures on dairy farms, exposing them to disease transmission. This survey also highlights regional and herd size-related differences in the implementation of biosecurity. Collecting data is an important first step to identification of specific weaknesses in different farm typologies, and an adequate follow-up is needed to ensure that measures are implemented correctly on farms.

Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq Bovine alphaherpesvirus 1 (BoHV-1) and Bovine viral diarrhea virus (BVDV) are important pathogens of cattle whose losses may be prevented by vaccination. This study evaluated the serological response induced by commercial vaccines and by an experimental vaccine associated with different adjuvants. For this, groups of seronegative heifers were immunized twice (30 days apart) with nine commercial vaccines and tested for virus neutralizing (VN) antibodies to the respective viruses 30 days after the second vaccine dose (D60). For the adjuvant test, inactivated BoHV-1 and BVDV-1 antigens were combined with six different adjuvants and submitted to the same immunization and serology protocol. Seven out of nine commercial vaccines induced VN antibodies to BoHV-1 at day D60, but only three induced titers considered protective (≥ 16). The frequency of seroconversion among the vaccines varied from 0% (0/10 animals) to 100% (10/10) and the geometric mean titers (GMT) ranged from 0 to 43. The frequency of seroconversion in protective titers was 100%, 37,5% and 25%, respectively. Only four vaccines induced neutralizing titers to BVDV-1 and three to BVDV-2 at D60. The frequency of seroconversion to BVDV among the vaccines varied from 0% (0/10 animals) to 100% (10/10) and the geometric mean titers (GMT) ranged from 0 to 202. Only three vaccines conferred titers compatible with protection (>60). The frequency of seroconversion in protective titers was 55,5%, 42,8% and 12,5% for BVDV-1, respectively. All experimental vaccine formulations induced VN titers against BoHV-1 at D60 (frequency of seroconversion varied from 0% to 100% and GMT= varied from 2 to 20) and four induced protective titers. The frequency of seroconversion in protective titers was 100%, 33,3%, 30% and 20%, respectively. On the other hand, none of the experimental vaccines was able to induce VN antibodies to BVDV at D60. Under the conditions of the present study, most commercial vaccines induced unsatisfactory VN response against the tested viruses, most noticeably to BVDV. Among the experimental vaccines, the adjuvant Emulsigen - DL90 (DDA) was more effective to induce VN antibodies against BoHV-1, but not to BVDV. These results indicate the need for reformulation of vaccine composition and/or immunization protocols as to achieve more consistent serological response to BoHV-1 and, especially, to BVDV. O alfaherpesvírus bovino 1 (BoHV-1) e o vírus da diarreia viral bovina (BVDV) são patógenos importantes em bovinos cujas perdas podem ser reduzidas pela vacinação. Este estudo avaliou a resposta sorológica induzida por vacinas comerciais e por uma vacina experimental inativada associada com diferentes adjuvantes. Para isso, grupos de novilhas foram imunizados duas vezes (com intervalo de 30 dias), com nove vacinas comerciais. Amostras de soro foram testadas pela técnica de neutralização viral (VN), para avaliar a indução de anticorpos neutralizantes para seus respectivos vírus, 30 dias após a segunda dose (D60). Para o teste de adjuvante, antígenos de BoHV-1 e BVDV-1 inativados foram combinados com seis adjuvantes diferentes e submetidos ao mesmo protocolo de imunização e sorologia. Sete de nove vacinas comerciais induziram anticorpos contra o BoHV-1 no dia D60, mas apenas três induziram títulos considerados protetores (≥ 16). A frequência de soroconversão entre as vacinas variou de 0% (0/10 dos animais) a 100% (10/10) e os títulos médios geométricos (GMT) variaram de 0 a 43. A frequência de soroconversão em títulos protetores foi de 100%, 37,5% e 25% para BoHV-1, respectivamente. Somente quatro vacinas induziram títulos neutralizantes contra o BVDV-1 e três ao BVDV-2 no D60. A frequência de soroconversão para o BVDV variou de 0% (0/10 animais) a 100% (10/10) e os títulos médios geométricos (GMT) variaram de 0 a 202. Apenas três vacinas conferiram títulos compatíveis com proteção (> 60). A frequência de soroconversão em títulos protetores foi de 55,5%, 42,8% e 12,5% para BVDV-1, respectivamente. Todas as formulações vacinais experimentais induziram títulos neutralizantes contra BoHV-1 no D60 (a frequência de soroconversão variou de 0% a 100% e o GMT variou de 2 a 20), mas apenas quatro induziram títulos protetores. A frequência de soroconversão em títulos protetores foi de 100%, 33,3%, 30% e 20%, respectivamente. Por outro lado, nenhuma das vacinas experimentais foi capaz de induzir anticorpos neutralizantes para BVDV no D60. Nas condições do presente estudo, a maioria das vacinas comerciais induziu resposta sorológica insatisfatória contra os vírus testados, sendo mais evidente para o BVDV. Entre as vacinas experimentais, o adjuvante Emulsigen - DL90 (DDA) foi o mais eficaz na indução de anticorpos contra o BoHV-1, mas não contra o BVDV. Estes resultados indicam a necessidade de reformulação da composição das vacinas e / ou dos protocolos de imunização, para obter resposta sorológica mais consistente para o BoHV-1 e, principalmente, para o BVDV.

RESUMO: Foram estudados 26 casos de meningoencefalite por herpesvírus bovino (BoHV) diagnosticados entre 2010-2016, no Estado de Goiás (GO). A doença acometeu principalmente bovinos jovens, entre 60 dias a 18 meses de idade. Não houve associação entre os casos e o sexo dos bovinos e a sazonalidade. A doença foi observada em todas as cinco Mesorregiões do Estado, com uma frequência maior nas Mesorregiões Sul e Centro. Os sinais clínicos mais frequentemente observados incluíram cegueira, incoordenação, sialorreia e ataxia. As principais alterações macroscópicas observadas incluíram congestão com tumefação e achatamento das circunvoluções, amolecimento e amarelamento do córtex telencefálico e focos de hemorragia. Em cinco encéfalos, não foram observadas alterações macroscópicas e em quatro as alterações não foram informadas. As principais alterações histológicas ocorreram no córtex telencefálico, principalmente o córtex frontal e parietal, mas em alguns casos, lesões de menor intensidade foram também observadas no tálamo, núcleos basais, mesencéfalo, ponte, bulbo, cerebelo e hipocampo. Todos os casos apresentaram meningoencefalite linfoplasmocítica e corpúsculos de inclusão intranucleares basofílicos em astrócitos e, eventualmente, em neurônios. Outras lesões frequentes incluíram necrose neuronal laminar segmentar (neurônio vermelho), espongiose, tumefação do núcleo das células endoteliais, gliose focal ou difusa, hipertrofia de astrócitos, infiltração por células gitter, congestão e hemorragia. Lesões menos comuns incluíram astrócitos Alzheimer tipo II, lesão residual e neuronofagia. A necrose neuronal e o edema (espongiose) foram mais acentuados nas camadas granular externa, molecular, de células piramidais e granular interna dos telencéfalos. Tanto os giros quanto os sulcos foram afetados igualmente. Dos 26 casos, o DNA de BoHV-5 foi amplificado em dois (7,69%) casos, enquanto que o de BoHV-1 foi identificado em um caso (3,84%). Nos casos positivos para BoHV-5 foram usadas amostras fixadas em formol a 10% e incluídas em parafina e amostras congeladas foram utilizadas no caso positivo para BoHV-1. ABSTRACT: Twenty six cases of bovine herpetic meningoencephalitis diagnosed from 2010-2016 in Goiás state, Brazil, were studied. Affected cattle were mainly 60-day to 18-month-old. There was no association of the disease with sex and seasonality. The disease was found in all five mesoregions with a higher prevalence in southern and central state of Goiás. Clinical signs more frequently observed included blindness, incoordination, circling, excessive salivation, and ataxia. Main gross findings in the brain were congestion with swelling and flattening of gyri, softening and yellow discoloration of cerebral cortex and hemorrhagic foci. In five cases no gross changes were observed in the brain and in four cases there is no information. The main histopathological changes were in the cortex of telencephalic lobes, especially the frontal and parietal; however less prominent and less frequently found lesions occurred in the thalamus, basal nuclei, midbrain, pons, medulla oblongata, cerebellum, and hippocampus. All cases presented lymphoplasmocytic meningoencephalitis and intranuclear basophilic inclusion bodies in astrocytes, less commonly in neurons. Other frequent lesions included segmental laminar neuronal necrosis (red neurons), spongiosis, swollen vascular endothelial nuclei, gliosis (focal and diffuse), hypertrophy of astrocytes, infiltration of gitter cells, congestion, and hemorrhage. Lesions less frequently observed were Alzheimer type II astrocytes, residual lesion and neuronophagia. The most frequently affected cortical layers by neuronal necrosis and edema were external and internal granular, molecular, and pyramidal cell layers. Gyri and sulci were equally affected. Of the 26 cases, in 2 (7.69%) the DNA of BoHV-5 was amplified with samples fixed in 10% formalin and paraffin-embedded. DNA of BoHV-1 was identified in another case (3.84%) where, positive to BoHV-1, fresh samples were used.

The study objectives were to: 1) examine how calves of divergent immune status respond to BRSV vaccination at 3 weeks of age; 2) trace glycoprotein E negative BoHV-1 antibodies from vaccinated dams to calf sera and to investigate how passive transfer affects response to live BoHV-1 vaccine at 6 weeks of age; 3) explore the impact of passive transfer status on blood metabolites around weaning. Thirty seven Holstein cows and their calves were included in the study. All cows were immunised with a commercial marker vaccine against BoHV-1(gE-) administered intra-muscularly at 4 month prior to the start of calving. Calves were assigned to 1 of 2 colostrum treatment groups: 1) 5% of BW in colostrum fed at birth, or 2) 10% of BW in colostrum fed at birth. Calves were also immunised at 3 weeks of age with a respiratory commercial vaccine, and a booster administered 4 weeks later. Calves were also immunised against BoHV-1 at 6 weeks of age, using one dose of a live commercial vaccine. The results demonstrated that level of passive immunity had no effect on immune response to vaccination and the importance of feeding colostrum from vaccinated BoHV-1 gE- dams to provide calves with passive protection against IBRV.

Bu çalışmanın amacı, ikili (Bovine viral diarrhea virus / Bovine herpesvirus -1) ve tek enfeksiyonlu (Bovine viral diarrhea virus veya Bovine herpesvirus -1) hayvanlarda ve ayrıca sağlıklı sığırlarda serum amiloid A, haptoglobin ve özellikle hepcidin konsantrasyonlarını belirlemekti. Daha önce sadece hepcidin geni üzerinde çalışmalar yapılmış olup ancak sığır hastalıklarında hepcidin değerleri araştırılmamıştır. Bu çalışmada toplam 56 sığır kullanılmıştır. Onlardan 17'inde Bovine herpesvirus-1, 13 tanesinde Bovine viral diarrhea virüsü, bunların 4'ünde ise hem Bovine herpesvirus-1 hem Bovine viral diarrhea virüsü tespit edildi. Kontrol grubu ise 22 sağlıklı sığırdan oluşturuldu. Bulgulara göre, çift infeksiyonlu hayvanlar ile tek enfeksiyonlu hayvanlar arasında akut faz proteinlerinin seviyelerinde belirgin farklılık saptanmamıştır. Serum hepcidin değerleri, 3 çalışma grubu arasında farklılık göstermedi. Fakat üç deneme grubunda kontrol grubuna göre anlamlı farklılıklar vardı (p

We aimed to determine the seroprevalence of infection with Neospora caninum, Leptospira, and bovine herpesvirus type 1 and risk factors associated with these infections in water buffaloes in Veracruz State, Mexico. Through a cross-sectional study, 144 water buffaloes (Bubalus bubalis) raised in 5 ranches of Veracruz were examined for anti-N. caninum and anti-bovine herpesvirus type 1 antibodies by enzyme immunoassays, and anti-Leptospira interrogans antibodies by microscopic agglutination test. Of the 144 buffaloes studied, 35 (24.3%) were positive for N. caninum, 50 (34.7%) for Leptospira, and 83 (57.6%) for bovine herpes virus. The frequencies of leptospiral serovars in buffaloes were as follows: 18.7% for Muenchen (n = 27), 10.4% for Hardjo LT (n = 15), 9.0% for Pyrogenes (n = 13), and 4.8% for Icterohaemorrhagiae (n = 7). Seropositive buffaloes were found in all 5 ranches studied. Logistic regression showed that cohabitation of buffaloes with cows was associated with infection with Leptospira (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.04–4.5; P = 0.03) and bovine herpesvirus (OR, 12.0; 95% CI, 4.0–36.2; P < 0.01). This is the first study that provides serological evidence of N. caninum, Leptospira, and bovine herpesvirus type 1 infections in water buffaloes in Mexico. Our findings could be used to enhance preventive measures against these infections.

This study aimed at comparing the surveillance program of bovine herpesvirus type 1 (BHV1) as laid down by EU Decision 2004/558/EC and 2007/584/EC (‘conventional design’) with an alternative design. The alternative design was based on monthly bulk-milk testing, clinical surveillance and a risk-based component that involves testing of animals that are purchased from non-free cattle herds. Scenario-tree analyses were carried out to determine sensitivities of the surveillance system (and its components) and the monthly confidence of freedom on herd-level. Also, the expected costs per surveillance design and components thereof were calculated. Results showed that the conventional (EU) and alternative surveillance designs to obtain a BHV1-free status performed equally well in terms of sensitivity. However, total costs per cattle herd to obtain a free status were highest in the conventional design. In an endemic situation and with a within-herd design prevalence of 10%, the conventional design led to a varying probability of freedom ranging from 99.6% to 100% per month. With the alternative design, in this situation, a constant probability of freedom of >99.9% per month was found. In a disease-free situation, both designs performed equally well (probability of freedom >99.9% per month). The yearly costs per farm for monitoring the disease-free status decreased by approximately 25% in the alternative design. The alternative strategy based on monthly bulk-milk monitoring therefore was deemed most cost-effective. This study showed that the surveillance regime to attain and maintain a BHV1-free status as described by EU-legislation can be improved to reduce the monitoring costs without reduction of the system’s sensitivity, given a within-herd design prevalence of 10%. The assessment of various surveillance designs could be highly useful to support decision-making towards a more risk-based approach of animal health surveillance.

Tesis para obtener el grado de Doctor en Ciencia Animal, presentada en la Universidad Nacional del Centro de la Provincia de Buenos Aires, en 2020 En la presente tesis se trabajó en invernadas del Valle 16 de Octubre, provincia de Chubut, con terneros de entre 6-8 meses de vida. Los objetivos fueron desarrollar conocimientos sobre la dinámica de circulación y características moleculares de Herpes Virus Bovino tipo 1 (BoHV1), Virus de Parainfluenza Bovina 3 (BPI3V) y Virus de la Diarrea Viral Bovina (BVDV), estudiar el rol de los mismos en la ocurrencia de casos clínicos de Complejo Respiratorio Bovino (CRB) y evaluar estrategias sanitarias para el control de CRB. Se realizaron 4 actividades. La primera fue evaluar el rol de posibles factores de riesgo correspondientes al animal y a medidas de manejo en la ocurrencia de casos clínicos de CRB. La segunda actividad se centralizó en conocer la dinámica de circulación de BoHV1, BPI3V y BVDV a partir de estudios serológicos y virológicos. La tercera se relacionó con la detección de casos clínicos de CRB y la evaluación del rol de los virus durante los mismos. Por último, se evaluaron diferentes esquemas de vacunación en bovinos vacunados con inmunógenos formulados a partir de aislamientos locales. No se identificaron factores de riesgo asociados a la ocurrencia de casos de CRB en este estudio, aunque el clima húmedo y frio característico de la zona durante el invierno, podría actuar como un potencial factor de riesgo. Los tres virus circularon en las invernadas estudiadas, BoHV1 lo hizo asociado a un cuadro de queratoconjuntivitis, BPI3V lo hizo desde el momento de ingreso a las invernadas y BVDV asociado a la mezcla de animales de diferentes orígenes. En los casos de CRB no se pudo confirmar que los virus estudiados hayan estado involucrados. Como estrategia de control se concluye que serían necesarias 2 dosis de vacuna para alcanzar niveles satisfactorios de inmunidad, la primera dosis puede ser aplicada al momento del ingreso de los terneros a las invernadas y la segunda dosis entre 21-28 días posteriores al mismo. In the present thesis was worked in feedlot of the Valley October 16, province of Chubut, with calves between 6-8 months of life. The objectives were to develop knowledge about the dynamics of circulation and molecular characteristics of Bovine Herpes Virus type 1 (BoHV1), Bovine Parainfluenza 3 Virus (BPI3V) and Bovine Viral Diarrhea Virus (BVDV), to study their role in the occurrence of clinical cases of Bovine Respiratory Complex (BRC) and evaluate sanitary strategies for the control of BRC. To achieve them, 4 activities were carried out. The first was to evaluate the role of possible risk factors for the animal and management measures in the occurrence of clinical cases of BRC. The second activity was focused on knowing the circulation dynamics of BoHV1, BPI3V and BVDV from serological and virological studies. The third was related to the detection of clinical cases of BRC and the evaluation of the role of the viruses during them. Finally, different vaccination schemes were evaluated in cattle vaccinated with immunogens formulated from local isolates. No risk factors associated with the occurrence of CRB cases were identified in this study, although the humid and cold climate characteristic of the area during winter could act as a potencial risk factor. The three viruses circulated in the feedlot periods studied, BoHV1 was associated with cases of keratoconjunctivites, BPI3V did so from the time of arrival to feedlot and BVDV associated with the commingled of calves from multiple sources. In the cases of BRC, it could not be confirmed that the viruses studied were involved. As a control strategy it is concluded that 2 doses of vaccine would be necessary to reach satisfactory levels of immunity, the first dose can be applied at the time of the arrival of the calves to the feedlot and the second dose between 21-28 days later. EEA Esquel Fil: Apóstolo, Romina María. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agroforestal Esquel; Argentina

The etiology of viral encephalitis in cattle often remains unresolved, posing a potential risk for animal and human health. In metagenomics studies of cattle with bovine non-suppurative encephalitis, parainfluenza virus 5 (PIV5) was identified in three brain samples. Interestingly, in two of these animals, bovine herpesvirus 6 and bovine astrovirus CH13 were additionally found. We investigated the role of PIV5 in bovine non-suppurative encephalitis and further characterized the three cases. With traditional sequencing methods, we completed the three PIV5 genomes, which were compared to one another. However, in comparison to already described PIV5 strains, unique features were revealed, like an 81 nucleotide longer open reading frame encoding the small hydrophobic (SH) protein. With in situ techniques, we demonstrated PIV5 antigen and RNA in one animal and found a broad cell tropism of PIV5 in the brain. Comparative quantitative analyses revealed a high viral load of PIV5 in the in situ positive animal and therefore, we propose that PIV5 was probably the cause of the disease. With this study, we clearly show that PIV5 is capable of naturally infecting different brain cell types in cattle in vivo and therefore it is a probable cause of encephalitis and neurological disease in cattle.

Bovine herpesvirus 1 (BoHV-1) is the causative agent of infectious bovine rhinotracheitis (IBR) and is also associated with reproductive failure. This study investigated the presence of BoHV-1 in cumulus-oocyte complexes (COCs) of naturally-infected cows without clinical signs of IBR. The presence of BoHV-1 in COCs was evaluated by immunofluorescence using confocal laser scanning microscopy. Blood samples and ovaries from 82 cows that had not been vaccinated against BoHV-1 were collected for serological analysis. COCs were divided into two pools: COCs derivate from seropositive cows and from seronegative cows. Then, the samples were processed for confocal microscopy analysis. The results indicated that 61% (50/82) of cows were seropositive for BoHV-1. A total of 719 COCs were obtained from the cows and processed. None of 276 COCs from the 32 seronegative cows presented BoHV-1. However, BoHV-1 was present in the cytoplasm of cumulus cells from 158 out of 443 COCs aspirated from the seropositive cows. The detection of BoHV-1 in the COCs of seropositive cows suggests that the COCs of naturally-infected, asymptomatic cows may be infected with BoHV-1.

Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are two closely related alpha-herpesviruses. BoHV-1 causes several syndromes in cattle, including respiratory disease and sporadic cases of encephalitis, whereas BoHV-5 is responsible for meningoencephalitis in calves. Although both viruses are neurotropic they differ in their neuropathogenic potential. This review summarizes the findings on the specific mechanisms and pathways known to modulate the pathogenesis of BoHV-1 and BoHV-5, particularly in relation with respiratory and neurological syndromes, which characterize the infections by each virus, respectively. Fil: Marin, Maia Solange. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Burucúa, Mercedes María. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rensetti, Daniel Ernesto. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Rosales Hurtado, Juan José. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Odeón, Anselmo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina Fil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina

Bovine herpesvirus type 1 (BoHV-1), a member of the Alphaherpesvirinae, causes a variety of diseases, which result in significant economic losses worldwide. Envelope glycoprotein D (gD) of BoHV-1 plays an important role in viral entry into the permissive cells, and protective immune response. The fine mapping epitope on the gD will contribute to the understanding of viral pathogenesis and development of alternative vaccines against various diseases associated with BoHV-1. We previously reported the preparation of a monoclonal antibody (MAb) 2B6, which was raised by a truncated recombinant gD protein, demonstrating a neutralizing activity against BoHV-1 infection in Madin-Darby bovine kidney cells. This study described the identification of a linear B-cell epitope on gD using MAb 2B6. A series of partially overlapping gD proteins with glutathione S-transferase tag were generated to define the epitope recognized by MAb 2B6. The amino acid (aa) sequence 323GEPKPGPSPDADRPE337 was recognized by MAb 2B6 using Western blot with the variedly truncated recombinant proteins. Importantly, this epitope was highly conserved among the typical members of BoHV-1, indicating that the epitope may be utilized in diagnosis of diseases due to BoHV-1 infection. Furthermore, the minimal linear epitope sequence 323GEPKPGP329 on gD recognized by MAb 2B6 was confirmed using single-aa residue deletion mutation in carboxyl terminal. This finding not only contributes to our understanding of gD of BoHV-1 virion but also shows a potential for the development of vaccine candidates and diagnostic techniques.

In this article, the results of molecular-genetic analyses of nasal swabs are presented. The experimental studies were carried out in the dairy farms of Vologda region suspected to the respiratory infections. The samples were obtained from the asymptomatic calves and from calves with cough and nasal discharges. The laboratory studies were conducted in the laboratory of virology of the FSC VIEV. The genome of the virus was detected by a polymerase chain reaction. The comparative analyses of identified nucleotide sequences of the virus with the reference strains of the International Nucleotide sequence database has shown the similarity with Bovine herpesvirus-5. Accordingly, as a result of our studies, the Bovine herpesvirus type 5 was identified in the nasal swabs of calves in farms of Vologda regions.

Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV) is a disease aff ecting cattle population of all breeds, categories and age. Th e disease can take diff erent clinical courses, infectious bovine rhinotracheitis (IBR) and infectious pustular vulvovaginitis (IPV) being the most common ones. Both diseases, an especially infectious bovine rhinotracheitis (IBR), pose severe health threat and cause major economic losses and are considered one of the most costly diseases in cattle industry. The causal agent is a virus categorized into the family Hepesviridae and designated as bovine herpesvirus type 1 (BHV-1) or IBR/IPV virus. Any positive result to IBR/IPV specifi c antibodies in non-vaccinated cattle, in either blood or milk, indicates the infection, and the animal is considered a source of infection. In this article, we described the needs and prospects for the eradication of IBR/IPV in the Republic of Serbia. The eradication of IBR/IPV is a complex process implying strictly defi ned program of measures. The implementation of such measures requires systematic strategy involving diff erent phases and activities that can continue over several years. The program requires substantial eff orts as well as fi nancial resources, which should be justifi ed and paid off through a successful eradication of IBR/IPV.

Bovine herpesvirus type 1 (BoHV-1) and bovine herpesvirus type 5 (BoHV-5) are important pathogens of cattle. The diseases they produce are quite different, with BoHV-5 being more neuropathogenic than BoHV-1 which mainly induces respiratory symptoms. The sequencing of the entire BoHV-5 genome has shown that most of the differences between these viruses are found in the immediate early and LR (latency related) genes. The LR gene is the only viral gene abundantly expressed in latently infected neurons, is essential for viral reactivation and seems to have an anti-apoptotic function which can be observed in vivo and in vitro. This gene spans two potential ORFs (1 and 2) which can also be found as a fused version, an ORF-E protein encoded within the promoter region and two miRNAs located within the 5’ UTR segment. Most of the essential functions of the LR gene seem to be located within the ORF-2 which has been found to modulate components of cell signaling/cycle pathways. In this review we present a comparative sequence analysis of the LR gene of several BoHV-5 isolates, their differences with the BoHV-1 homologue and the potential impact this may have on its function. The LR gene was found to be highly conserved in all sequenced BoHV-5 strains. ORF-1 shares 60 % homology compared to BoHV-1 whereas the BoHV-5 homologue of ORF-2 is truncated at amino acid 51. Preliminary studies analyzing the emerging transcripts from the BoHV-5 LR gene in infected cells, as well as in stably transfected cells, indicates that their products are, in fact, missing crucial components of the anti-apoptotic function when compared to the BoHV-1 LR gene. In addition these transcripts maintain a region that, similar to what is found in BoHV-1, would produce a miRNA with the potential to recognize a region within the BoHV-5 immediate early gene. All together, these BoHV-5 characteristics suggest that this virus would not possess the same repertoire of latency maintaining functions as BoHV-1. Implications for BoHV-5 neuropathogenic potential are discussed. Fil: Silvestro, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento Virologia; Argentina Fil: Bratanich, Ana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento Virologia; Argentina

The objective of this work was to investigate the presence of bovine herpesvirus type 1 (BoHV-1) in follicular fluid and in cumulus-oocyte complexes (COC) recovered from naturally infected cows but with no clinical signs of the disease. Cows that were seropositive (n=38) or seronegative (n=8, control) to infectious bovine rhinotracheitis were selected after a serum neutralization test in microplates. The presence of the virus was investigated by PCR in COC and in follicular fluid. Viral DNA was not found in any of the samples. The obtained results suggest that serologically positive cows with no clinical signs of the disease offer negligible risk of transmitting BoHV-1 by COC or follicular fluid. Resumo: O objetivo deste trabalho foi investigar a presença do herpesvírus bovino tipo 1 (BoHV-1) no fluido folicular e em complexos cumulus-oócito (COC) coletados de vacas naturalmente infectadas mas sem sinais clínicos da doença. Vacas soropositivas (n=38) ou soronegativas (n=8, controle) para o vírus da rinotraqueíte infecciosa bovina foram selecionadas por prova de soroneutralização em microplacas. A presença do vírus foi investigada por PCR em COC e fluido folicular. O DNA viral não foi encontrado em qualquer das amostras. Os resultados obtidos são indicativos de que vacas soropositivas, mas sem sinais clínicos da doença, oferecem risco negligível de transmissão do BoHV-1 por COC ou fluido folicular.

Crimean-Congo hemorrhagic fever virus (CCHFV) is the causative agent of a tick-borne infection with significant mortality rate of up to 40% in the endemic areas, with evidence for geographical expansion. Lacking effective therapeutics and control measures, the development of protective CCHFV vaccine remains a crucial public health task. This manuscript describes, for the first time, a Bovine herpesvirus type 4 (BoHV-4) based viral vector (BoHV4-∆TK-CCHFV-N) and its immunogenicity and protection potential in BALB/c and IFNAR-/- mice models in comparison with Adenovirus type 5 (Ad5-N) and pCDNA3.1 myc/His A (pCD-N1), two widely used vaccine platforms. All constructs expressing viral nucleocapsid (N) protein successfully elicited cytokine and total/specific antibody responses in BALB/c mice. BoHV4-∆TK-CCHFV-N and Ad5-N constructs further produced 100% protection in IFNAR-/- mice during CCHFV Ank-2 strain lethal challenge. Despite elevated specific antibody responses in both animal models, the produced antibodies were unable to neutralize the virus in vitro. A comparison of delivery platforms was not possible, due to similar protection rates in IFNAR-/- mice. In conclusion, vector-based CCHFV N protein expression proved to constitute an effective approach for the vaccine development pipeline and BoHV-4 emerged as a strong alternative to previously-used virus vectors.

Bovine herpesvirus 1 (BoHV-1) is one of several microbes that contributes to the development of the bovine respiratory disease (BRD) and can also induce abortions in cattle. As other alpha-herpesvirinae subfamily members, BoHV-1 efficiently replicates in many cell types and subsequently establishes a life-long latent infection in sensory neurons. BoHV-1 encodes more than 70 proteins that are expressed in a well-defined manner during productive infection. However, in silico open reading frame (ORF) prediction of the BoHV-1 genome suggests that the virus may encode more than one hundred proteins. In this study we used mass spectrometry followed by proteogenomic mapping to reveal the existence of 92 peptides that map to previously un-annotated regions of the viral genome. Twenty-one of the newly termed &ldquo, intergenic peptides&rdquo, were predicted to have a viable ORF around them. Twelve of these produced an mRNA transcript as demonstrated by strand-specific RT-PCR. We further characterized the 5&prime, and 3&prime, termini of one mRNA transcript, ORF-A, and detected a 55 kDa protein produced during active infection using a custom-synthesized antibody. We conclude that the coding potential of BoHV-1 is underestimated.

BoHV-1 genera en el ganado bovino una amplia gama de manifestaciones clínicas, tales como la rinotraqueítis infecciosa bovina, abortos y vulvovaginitis pustular infecciosa, que dependen de la virulencia del virus y del estado inmunitario del hospedador. En este trabajo se caracterizaron en Argentina 44 aislamientos de BoHV-1 por el análisis de restricción con endonucleasas (REA). El ADN viral fue sometido al clivaje de la enzima HindIII, la cual permite la diferenciación entre los subtipos 1.1, 1.2a y 1.2b. De acuerdo al perfil REA de los aislamientos, 35 cepas (80%) mostraron un patrón similar a la cepa K22 (prototipo BoHV- 1.2b), 7 cepas (16%) exhibieron un patrón similar a la cepa LA (prototipo BoHV-1.1) y sólo 2 cepas (4%) revelaron un patrón similar a la cepa ST (prototipo BoHV-1.2a). Los resultados demuestran una mayor prevalencia del subtipo 1.2b entre los aislamientos BoHV-1 argentinos. Además, se reporta por primera vez en Argentina, la circulación del subtipo BoHV-1.2a. BoHV-1 in cattle results in a wide range of clinical manifestations, such as infectious bovine rhinotracheitis, abortion and infectious pustular vulvovaginitis, depending on the virulence of the virus and the immune status of the host. Forty-four Argentinean BoHV1 isolates were characterized by restriction endonuclease analysis (REA). Viral DNA was cleaved with HindIII which allows the differentiation among subtypes 1.1, 1.2a and 1.2b. According to the REA, 35 field strains (80%) showed a pattern similar to K22 strain (BoHV-1.2b prototype), 7 field strains (16%) showed a clear pattern similar to LA strain (BoHV-1.1 prototype) and only 2 filed strains (4%) showed a clear pattern similar to ST strain (BoHV-1.2a prototype). The results demonstrated the presence of a marked prevalence of subtype 1.2b among Argentinean BoHV-1 isolates. We also report for the first time in Argentina the circulation of the subtype BoHV-1.2a. BoHV-1 in cattle results in a wide range of clinical manifestations, such as infectious bovine rhinotracheitis, abortion and infectious pustular vulvovaginitis, depending on the virulence of the virus and the immune status of the host. Forty-four Argentinean BoHV1 isolates were characterized by restriction endonuclease analysis (REA). Viral DNA was cleaved with HindIII which allows the differentiation among subtypes 1.1, 1.2a and 1.2b. According to the REA, 35 field strains (80%) showed a pattern similar to K22 strain (BoHV-1.2b prototype), 7 field strains (16%) showed a clear pattern similar to LA strain (BoHV-1.1 prototype) and only 2 filed strains (4%) showed a clear pattern similar to ST strain (BoHV-1.2a prototype). The results demonstrated the presence of a marked prevalence of subtype 1.2b among Argentinean BoHV-1 isolates. We also report for the first time in Argentina the circulation of the subtype BoHV-1.2a. Instituto de Virología Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Morón. Facultad de Ciencias Exactas, Químicas y Naturales. Cátedra de Inmunogenética; Argentina Fil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina Fil: De Stefano, Gabriel Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Combessies, Gustavo. Laboratorio Azul; Argentina Fil: Romera, Sonia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Morón. Facultad de Ciencias Exactas, Químicas y Naturales. Cátedra de Inmunogenética; Argentina. Universidad del Salvador. Cátedra de Inmunología; Argentina

Oncolytic viruses selectively replicate in tumor cells and elicit antitumor effects in vivo by both direct and indirect methods. They are attractive avenues of cancer therapy due to the absence of toxic side effects often seen in current treatment modalities. Bovine herpesvirus type 1 (BHV-1) holds promise as a broad-spectrum oncolytic vector that is able to infect and kill human tumor cells from a variety of histological origins, including cancer-initiating cells. In the majority of cases, BHV-1 elicits tumor cell death in the absence of a productive infection. In vivo, BHV-1 affects the incidence of secondary lesions in cotton rats bearing subcutaneous breast adenocarcinomas. These recent studies contribute to the characterization of BHV-1 as an oncolytic virus.

In this work, the photodynamic efficiency of anionic meso-tetrakis sulfonophenyl (TPPS4), cationic meso-tetrakis methylpyridiniumyl (TMPyP) and their zinc complexes (ZnTPPS4 and ZnTMPyP) in the inactivation of Bovine herpesvirus type 1 (BoHV-1) was evaluated. At a non-cytotoxic concentration, all porphyrins showed significant antiviral activity after irradiation using a halogen lamp. The efficiency of the cationic porphyrins was higher than that of the anionic ones. Porphyrin complexation with zinc increases its lipophilicity and the number of absorbed photons, dramatically reducing the time for complete virus inactivation. The high superposition of the compound optical absorption and light source emission spectra played a key role in the virus inactivation efficiency. The results demonstrated the high effectivity of the photodynamic inactivation of BoHV-1. This method can be used as an auxiliary in the treatment of disorders attributed to BoHV-1 infection, and the porphyrins are promising photosensitizers for this application.

Background Bovine Viral Diarrhea Virus (BVDV) and Bovine Herpesvirus type 1 (BoHV-1) cause reproductive problems in cattle and restrictions on international trade in animals worldwide. Both infections were detected in cattle herds in the Paraíba state, Northeastern Brazil, however, the spatial distribution and geographic identification of positive herds for these viruses has never been examined. Therefore, the aim of this study was to describe the spatial pattern of apparent prevalence estimate and to identify spatial clustering of positive herds of BVDV and BoHV-1 infections in cattle herds from the state of Paraíba, Northeastern Brazil. Results The herd-level prevalence for BVDV and BoHV-1 infections in Paraíba were, respectively, 65.5% (95% CI: 61.1–69.7) and 87.8% (95% CI: 84.5–90.5). The average apparent within-herd prevalence of BVDV was 31.8% and of BoHV-1 was 62.4%. The predicted prevalence was highest (0.42–0.75) for BVDV in the west, north and eastern part of Sertão and in the central and eastern part of Agreste/Zona da Mata. For BoHV-1, the highest predicted prevalence (0.74–0.97) was in some local areas across Sertão and throughout the eastern part of Agreste/Zona da Mata. Six significant clusters were detected for BVDV, a primary cluster covering the eastern Sertão region, with 11 herds, radius of 24.10 km and risk relative (RR) of 2.21 (P

The aims of this study were to determine serum amyloid A, haptoglobinand especially hepcidin concentrations in animals with dual (Bovineviral diarrhea virus / Bovine herpesvirus-1)and single infections ( Bovineviral diarrhea virus or Bovine herpesvirus -1) andreference ranges of hepcidin in healthy clinicaly cattle. Previously, studies have been conducted on only hepcidin genebut hepcidin values has not been studied in cattle diseases. In thisstudy totaly 56 cattle were used. In 17 of them Bovine herpesvirus-1,13 of them Bovine viral diarrhea virus,4 of them both Bovine herpesvirus-1 and Bovineviral diarrhea virus were diagnosed. The control group wascomposed of 22 clinicaly healthy animals. According to the results animals naturally infected with dual infection did notshow more significant differences level of acute phase proteins compared to animals with single infection. Serum hepcidin values​​did not differ between the 3 trialgroups. But there were significant differences in three trialgroups results as compared to control group (p, Buçalışmanın amacı, ikili (Bovine viral diarrhea virus / Bovine herpesvirus -1)ve tek enfeksiyonlu (Bovine viral diarrhea virus veya Bovine herpesvirus -1)hayvanlarda ve ayrıca sağlıklı sığırlarda serum amiloid A, haptoglobin veözellikle hepcidin konsantrasyonlarını belirlemekti. Dahaönce sadece hepcidin geni üzerinde çalışmalar yapılmış olup ancak sığırhastalıklarında hepcidin değerleri araştırılmamıştır.Buçalışmada toplam 56 sığır kullanılmıştır. Onlardan 17'inde Bovine herpesvirus-1,13 tanesinde Bovine viral diarrhea virüsü, bunların 4'ünde ise hem Bovine herpesvirus-1hem Bovine viral diarrhea virüsü tespit edildi. Kontrol grubu ise 22 sağlıklı sığırdanoluşturuldu. Bulgulara göre, çift infeksiyonlu hayvanlar ile tekenfeksiyonlu hayvanlar arasında akut faz proteinlerinin seviyelerinde belirginfarklılık saptanmamıştır. Serum hepcidin değerleri, 3 çalışmagrubu arasında farklılık göstermedi. Fakat üç deneme grubunda kontrol grubunagöre anlamlı farklılıklar vardı (p

Se describe un brote IBR con aislamiento de Herpes Virus Bovino tipo 1 (BoHV1) en la Patagonia argentina. Durante los meses de junio-julio de 2013 tuvo lugar un brote de rinitis y conjutivitis en un lote de terneros confinados en una recría a corral en el Valle 16 de Octubre, al noroeste de la provincia de Chubut. Debido a que los síntomas eran compatibles con la infeccion por BoVH1, se procedió a tomar muestras de secreciones oculares y nasales de 12 animales y muestras de suero de 7 animales. A partir de las muestras de secreciones se lograron aislar 10 cepas de BoHV1. Si bien el hallazgo no permite concluir sobre el rol del BoHV1 en el brote, constituye el primer reporte de aislamiento de BoVH1 y confirma los trabajos previos sobre evidencia serológica de su circulación en la Patagonia argentina. An outbreak of IBR with the isolation of Bovine Herpes Virus type 1 (BoHV1) in Argentinean Patagonia is described. During June-July 2013, an outbreak of rhinitis and conjunctivitis occurred in a group of calves confined to a farm in 16 de Octubre Valley, northwest of Chubut. Because the symptoms were compatible with the infection with BoHV1, samples of ocular and nasal secretions from 12 animals and serum samples from 7 animals were collected. Ten strains of BoHV1 from the secretion samples were isolated. Although the finding does not allow to conclude about the role of BoHV1 in the outbreak, it constitutes the first report of isolation of BoHV1 and confirms previous studies on serological evidence of its circulation in Argentinean Patagonia. Estación Experimental Agropecuaria Bariloche Fil: Apostolo, Romina Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Esquel. Grupo de Producción Animal; Argentina Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituo de Investigación Virología; Argentina Fil: Martinez Stanziola, Juan Pablo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Esquel. Agencia de Extensión Rural Trevelin; Argentina Fil: Romera, Sonia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituo de Investigación Virología; Argentina Fil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Area de Produccion Animal. Grupo de Sanidad Animal; Argentina

Bovine herpesvirus (BoHV) type 1 is an important agricultural pathogen that infects cattle and other ruminants worldwide. Acute infection of the oro-respiratory tract leads to immune suppression and allows commensal bacteria to infect an otherwise healthy lower respiratory tract. This condition is known as the Bovine Respiratory Disease (BRD). BoHV-1 latently infects the host for life and periodical stress events re-initiate BRD, translating into high morbidity and large economic losses. To gain a better understanding of the biology of BoHV-1 and the disease it causes, we elucidated the protein composition of extracellular virions using liquid chromatography-mass spectrometry analysis. We detected 33 viral proteins, including the expected proteins of the nucleocapsid and envelope as well as other regulatory proteins present in the viral tegument. In addition to viral proteins, we have also identified packaged proteins of host origin. This constitutes the first proteomic characterization of the BoHV virion.

A thymidine kinase (tk)-deleted bovine herpesvirus 5 (BoHV-5tkΔ) was previously shown to establish latent infection and reactivate - even poorly - in a sheep model (Cadore et al. 2013). As TK-negative alphaherpesviruses are unlike to reactivate in neural tissue, this study investigated the sites of latency and reactivation by this recombinant in lambs. For this, groups of lambs were inoculated intranasally with the parental BoHV-5 strain (SV-507/99) or with the recombinant BoHV-5tkΔ. During latent infection (40 days post-inoculation, pi), the distribution of recombinant virus DNA in neural and non-neural tissues was similar to that of the parental virus. Parental and recombinant virus DNA was consistently detected by PCR in trigeminal ganglia (TGs); frequently in palatine and pharyngeal tonsils and, less frequently in the retropharyngeal lymph nodes. In addition, latent DNA of both viruses was detected in several areas of the brain. After dexamethasone (Dx) administration (day 40pi), the recombinant virus was barely detected in nasal secretions contrasting with marked shedding of the parental virus. In tissues of lambs euthanized at day 3 post-Dx treatment (pDx), reverse-transcription-PCR (RT-PCR) for a late viral mRNA (glycoprotein D gene) demonstrated reactivation of parental virus in neural (TGs) and lymphoid tissues (tonsils, lymph node). In contrast, recombinant virus mRNA was detected only in lymphoid tissues. These results demonstrate that BoHV-5 and the recombinant BoHV-5tkΔ do establish latent infection in neural and non-neural sites. Reactivation of the recombinant BoHV-5tkΔ, however, appeared to occur only in non-neural sites. In anyway, the ability of a tk-deleted strain to reactivate latent infection deserves attention in the context of vaccine safety.

Bovine Herpesvirus type 5 (BoHV-5) has not been conclusively demonstrated to cause bovine abortion. Brain lesions produced by Neospora caninum and Bovine Herpesvirus type 1 (BoHV-1) exhibit common features. Therefore, careful microscopic evaluation and additional diagnostic procedures are required to achieve an accurate final etiological diagnosis. The aim of the present work was to investigate the occurrence of infections due to BoHV-1, BoHV-5 and N. caninum in 68 cases of spontaneous bovine abortions which showed microscopic lesions in the fetal central nervous system. This study allowed the identification of 4 (5.9%) fetuses with dual infection by BoHV-5 and N. caninum and 33 (48.5%) cases in which N. caninum was the sole pathogen identified. All cases were negative to BoHV-1. The results of this study provide evidence that dual infection by BoHV-5 and N. caninum occur during pregnancy in cattle; however, the role of BoHV-5 as a primary cause of bovine abortion needs further research. Molecular diagnosis of BoHV-5 and N. caninum confirmed the importance of applying complementary assays to improve the sensitivity of diagnosing bovine abortion. Não está demonstrado até ao momento, que o Herpesvírus bovino tipo 5 (BoHV-5) seja um agente causal de aborto bovino. Uma vez que as lesões cerebrais tanto de Neospora caninum como de Herpesvírus bovino tipo 1(BoHV-1) têm características similares, é necessária uma avaliação microscópica cuidadosa, bem como exames laboratoriais adicionais, para obter um diagnóstico final preciso. O objetivo do presente trabalho foi investigar a presença de infeções por BoHV-1, BoHV-5 e N. caninum em 68 casos de aborto espontâneo, nos quais se verificaram lesões microscópicas no sistema nervoso central. Foram encontrados 4 (5,9%) fetos com infeção simultânea de BoHV-5 e N. caninum e 33 (48,5%) casos com infeção exclusiva de N. caninum. Todos os casos foram negativos a BoHV-1. Os resultados deste estudo demonstram que a infeção dual por BoHV-5 y N. caninum está presente durante a gestçao dos bovinos. Apesar disso, o papel de BoHV-5 como agente primário causal de aborto, carece de mais investigaçao. O diagnóstico molecular de BoHV-5 e N. caninum confirmou a importância de se aplicar ensaios complementares para melhorar a sensibilidade do diagnóstico de aborto bovino.

AVPDS (Animal Viruses Probe dataset) is a dataset of virus-specific and conserve oligonucleotides for identification and diagnosis of viruses infecting animals. The current dataset contain 20,619 virus specific probes for 833 viruses and their subtypes and 3,988 conserved probes for 146 viral genera. Dataset of virus specific probe has been divided into two fields namely virus name and probe sequence. Similarly conserved probes for virus genera table have genus, and subgroup within genus name and probe sequence. The subgroup within genus is artificially divided subgroups with no taxonomic significance and contains probes which identifies viruses in that specific subgroup of the genus. Using this dataset we have successfully diagnosed the first case of Newcastle disease virus in sheep and reported a mixed infection of Bovine viral diarrhea and Bovine herpesvirus in cattle. These dataset also contains probes which cross reacts across species experimentally though computationally they meet specifications. These probes have been marked. We hope that this dataset will be useful in microarray-based detection of viruses. The dataset can be accessed through the link https://dl.dropboxusercontent.com/u/94060831/avpds/HOME.html.

Bovine herpesvirus type 1 (BoHV-1) is the causative agent for infectious bovine rhinotracheitis and infectious pustular vulvovaginitis in cows or balanoposthitis in bulls. In this study, individual and bulk tank milk (BTM) samples from 5 Catalan dairy farms with different control strategies against BoHV-1 were analysed during the course of a year for milk quality parameters and glycoprotein E (gE) antibodies. Detection of gE antibodies was carried out with ELISA techniques. Prevalence of BoHV-1 varied between farms, and was stable during the study in individual and BTM samples. Comparing the antibody results of samples with milk quality parameters, positive samples with higher levels of antibodies corresponded to lower lactose and to higher percentages of fat and somatic cells.

Although bovine herpesvirus type 4 (BoHV-4) is primarily associated with reproductive disorders of cattle, it canproduce a variety of clinical signs. To determine the distribution, the presence and type of microscopic lesions causedby BoHV-4 strains of different genotypes an in vivo model, calves were infected with three phylogenetically differentArgentinean BoHV-4 strains. Samples from nasal and ocular secretions, peripheral blood leukocytes, tissues andserum were analyzed. BoHV-4 was isolated from nasal and ocular secretions at 7 and 14 days post-inoculation (dpi).Viral DNA was detected by nested PCR in peripheral blood leukocytes at 14 and 21 dpi for two out of three strainsand in tissues, such as nervous system, trachea, pulmonary and retropharyngeal lymph nodes, spleen and kidney, at21 dpi. Antibody levels detected by viral seroneutralization test were mostly low and varied widely for the differentstrains. The tissue distribution of the BoHV-4 strains and the variations observed in the levels of neutralizing antibodiesindicate that certain differences can be established among the patterns of biological behavior of each strain.This is an initial step to get insight into the biological characteristics of Argentinean BoHV-4 isolates. However,further evaluation involving a higher number of inoculated animals will be required to be conclusive on this aspect. Fil: Morán, Pedro Edgardo. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Virologia; Argentina Fil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Virologia; Argentina Fil: Garcia, Jo. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Virologia; Argentina Fil: Castro, Alejandra Nelly Cristina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Virologia; Argentina Fil: Spetter Lucas, Maximiliano Joaquín. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Odeón, Anselmo Carlos. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Virologia; Argentina Fil: Verna, Andrea Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina

Bovine herpesvirus type 5 (BoHV-5) is an important pathogen that causes meningoencephalitis in cattle. Few studies have used the mouse as a model for BoHV-5 infection. Despite the fact that BoHV-5 can infect mice with immune deficiencies, little is known about viral replication, immune response, and the course of infection in the central nervous system (CNS) of wild-type mice. Therefore, the aim of this study was to evaluate the response in the CNS of BALB/c mice acutely infected with BoHV-5 at different days post-inoculation (dpi). BoHV-5, when inoculated intracranially, was able to infect and replicate within the CNS of BALB/c mice. Until 15 dpi, the mice were able to survive without showing prominent neurological signs. The infection was accompanied by a Th1 immune response, with a significant expression of the cytokines IFN-γ and TNF-α and chemokine CCL-2. The expression of these cytokines and chemokines was most significant in the early course of infection (3 and 4 dpi), and it was followed by meningoencephalitis with perivascular cuffing and periventriculitis, composed mainly of macrophages and lymphocytes. After the expression of cytokines and chemokine, the mice were able to curb BoHV-5 acute infection in the brain, since there was a decrease in the number of BoHV-5 DNA copies after 3 dpi and viable viral particles were not detected after 6 dpi. Importantly, BoHV-5 was able to infect the trigeminal ganglia during acute infection, since a large number of BoHV-5 DNA copies were detected on 1 and 2 dpi.

Agaricus brasiliensis is an edible mushroom, traditionally used for the treatment of several diseases. In this paper, a polysaccharide (PLS) from A. brasiliensis, its carboxymethylated (CPLS) and sulfated (SPLS) derivatives, as well as, fractions (F1–F3) obtained from the PLS were investigated for their effect in the replication of herpes simplex virus and bovine herpes virus in HEp-2 cell cultures. The PLS, SPLS and F3 inhibited both virus strains similarly, in a dose-dependent curve. F1, F2 and CPLS did not show significant effect even at higher concentrations. All the compounds showed neither virucidal or viral adsorption inhibition activities nor effect when cells were treated prior to infection. Our study demonstrated that the extracts of A. brasiliensis, can be promising for future antiviral drug design and its biotechnological production is economically feasible.

This article describes the expression of a truncated form of bovine herpesvirus 1 (BoHV-1) glycoprotein E (gE) for use as immunodiagnostic reagent. A 651 nucleotide fragment corresponding to the amino-terminal third (217 amino acids) of BoHV-1 gE - that shares a high identity with the homologous BoHV-5 counterpart - was cloned as a 6×His-tag fusion protein in an Escherichia coli expression vector. A soluble protein of approximately 25 kDa purified from lysates of transformed E. coli was recognized in Western blot (WB) by anti-6xHis-tag and anti-BoHV-1 gE monoclonal antibodies. In addition, the recombinant protein was specifically recognized in WB by antibodies present in the sera of cattle seropositive to BoHV-1 and BoHV-5. An indirect ELISA using the expressed protein as coating antigen performed comparably to a commercial anti-gE ELISA and was able to differentiate serologically calves vaccinated with a gE-deleted BoHV-5 strain from calves infected with BoHV-1. Thus, the truncated gE may be useful for serological tests designed to differentiate BoHV-1/BoHV-5 infected animals from those vaccinated with gE-negative marker vaccines. Este trabalho relata a expressão de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 (BoHV-1) para uso em imunodiagnóstico. Um fragmento de 651 pares de bases (pb) correspondente ao terço amino-terminal (217 aminoácidos) da gE do BoHV-1 - que compartilha uma alta identidade com a gE do BoHV-5 - foi clonada como proteína de fusão com cauda 6x de histidina em um vetor de expressão em Escherichia coli. Uma proteína solúvel de aproximadamente 25 kDa purificada de lisados de E.coli foi reconhecida em Western blot (WB) por anticorpos monoclonais anti-6xHis-tag e anti-gE. Além disso, a proteína recombinante purificada foi reconhecida em WB por anticorpos presentes no soro de animais soropositivos ao BoHV-1 e BoHV-5. Um ELISA indireto utilizando a proteína recombinante como antígeno apresentou performance comparável a um ELISA gE comercial e foi capaz e diferenciar sorologicamente animais vacinados com uma cepa gE-negativa de BoHV-5 de animais infectados com o BoHV-1. Portanto, a gE truncada pode ser útil em testes sorológicos diferenciais para uso conjunto com vacinas com marcador antigênico gE para o BoHV-1 e BoHV-5.

This article describes the expression of a truncated form of bovine herpesvirus 1 (BoHV-1) glycoprotein E (gE) for use as immunodiagnostic reagent. A 651 nucleotide fragment corresponding to the amino-terminal third (217 amino acids) of BoHV-1 gE - that shares a high identity with the homologous BoHV-5 counterpart - was cloned as a 6×His-tag fusion protein in an Escherichia coli expression vector. A soluble protein of approximately 25 kDa purified from lysates of transformed E. coli was recognized in Western blot (WB) by anti-6xHis-tag and anti-BoHV-1 gE monoclonal antibodies. In addition, the recombinant protein was specifically recognized in WB by antibodies present in the sera of cattle seropositive to BoHV-1 and BoHV-5. An indirect ELISA using the expressed protein as coating antigen performed comparably to a commercial anti-gE ELISA and was able to differentiate serologically calves vaccinated with a gE-deleted BoHV-5 strain from calves infected with BoHV-1. Thus, the truncated gE may be useful for serological tests designed to differentiate BoHV-1/BoHV-5 infected animals from those vaccinated with gE-negative marker vaccines.Este trabalho relata a expressão de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 (BoHV-1) para uso em imunodiagnóstico. Um fragmento de 651 pares de bases (pb) correspondente ao terço amino-terminal (217 aminoácidos) da gE do BoHV-1 - que compartilha uma alta identidade com a gE do BoHV-5 - foi clonada como proteína de fusão com cauda 6x de histidina em um vetor de expressão em Escherichia coli. Uma proteína solúvel de aproximadamente 25 kDa purificada de lisados de E.coli foi reconhecida em Western blot (WB) por anticorpos monoclonais anti-6xHis-tag e anti-gE. Além disso, a proteína recombinante purificada foi reconhecida em WB por anticorpos presentes no soro de animais soropositivos ao BoHV-1 e BoHV-5. Um ELISA indireto utilizando a proteína recombinante como antígeno apresentou performance comparável a um ELISA gE comercial e foi capaz e diferenciar sorologicamente animais vacinados com uma cepa gE-negativa de BoHV-5 de animais infectados com o BoHV-1. Portanto, a gE truncada pode ser útil em testes sorológicos diferenciais para uso conjunto com vacinas com marcador antigênico gE para o BoHV-1 e BoHV-5.

Modified-live virus (MLV) bovine herpesvirus 1 (BHV-1) vaccines have been a major part of the program to the prevention of reproductive disease in cattle. However, their use (or misuse) continues to result in major abortion storms. The results of 20 additional cases with reproductive losses are described where the single nucleotide polymorphism (SNP) analysis was done to identify the source., American Association of Bovine Practitioners Proceedings of the Annual Conference, 2016

Objective . The aim of this study was to determine the presence of Bovine herpesvirus - 1 (BoHV-1) and Bovine herpesvirus 5 (BoHV-5) neutralizing antibodies in herds on the Colombian high plains and their correlation with the level of cross-protection against both herpesviruses. Materials and methods . This study was carried out on cattle farms located around the towns of Puerto López and Puerto Gaitán in Colombia's Meta department. Sampling was made by convenience. Twenty-three farms were involved in the study; 488 sera samples were taken by random sampling. Virus neutralization test were performed according to the protocols of the OIE. Each serum was evaluated independently for each virus, BoHV-1 and BoHV-5. Results. The serological test confirmed the presence of BoHV-1 and BoHV-5 infections in the Colombian bovine population in 100% and 73.9% respectively. However, cross-reaction for both viruses was not evident in all farms evaluated. Conclusions . Alpha-herpesviruses are amongst the most significant infectious agents affecting cattle. Bovine herpesvirus 1 (BoHV-1) is found throughout the whole world and is endemic in Colombian bovine population, whereas bovine herpesvirus 5 (BoHV-5) has limited geographical distribution, mainly being reported in South-America (Brazil and Argentina), and we also confirmed the presence of BoHV-5 in Colombia. RESUMEN Objetivo. El objetivo de este estudio fue determinar la presencia de anticuerpos para herpesvirus bovino 1 (BoHV-1) y herpesvirus bovino 5 (BoHV-5), así como también determinar el nivel de protección cruzada contra los dos herpesvirus en hatos de la altillanura colombiana. Materiales y métodos . Este estudio se realizó en fincas ubicadas en los municipios de Puerto López y Puerto Gaitán departamento del Meta. Se realizó un muestreo por conveniencia. Se emplearon 23 fincas, donde se realizó un muestreo al azar en cada una para un total de 488 muestras. Las muestras de suero fueron analizadas mediante sero-neutralización viral empleando los protocolos de la OIE. Cada uno de los sueros fue evaluado de manera independiente para cada uno de los virus BoHV-1 y BoHV-5. Resultados. Mediante la sero-neutralización viral se confirmó la presencia de BoHV-1 y BoHV-5 en la población bovina colombiana en 100% y 73.9%, respectivamente. Sin embargo, la reactividad cruzada no fue evidente en todas las fincas evaluadas. Conclusiones. Los alfa-herpesvirus bovinos se encuentran entre los agentes infecciosos más importantes del ganado bovino. Particularmente, el herpesvirus bovino 1 (BoHV-1) presenta una distribución mundial, siendo endémico en la población bovina colombiana, mientras que el herpesvirus bovino 5 (BoHV-5), presenta una distribución geográfica limitada; reportándose principalmente en Suramérica (Brasil y Argentina), comprobándose la presencia de este en los hatos colombianos.

Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are closely related alpha-herpesviruses. BoHV-5 is the causal agent of non-suppurative meningoencephalitis in calves. BoHV-1 causes respiratory disease, abortions, genital disorders and, occasionally, encephalitis in cattle. Both viruses are neurotropic and they share similar biological properties. Nevertheless, they differ in their ability to cause neurological disease. Toll-like receptors (TLRs) are involved in the innate immune response to pathogens. In this study, the variations in the expression levels of TLRs were evaluated in different regions of the bovine central nervous system during the acute infection and reactivation of BoHV-1 and BoHV-5- infected cattle. With the exception of TLR9, significant up-regulation of all TLRs was detected following primary infection of neural tissues by both bovine alpha-herpesviruses. Furthermore, the stages of acute infection and reactivation were characterized by a distinguishable TLR expression pattern. Important differences in TLR expression upon infection of the central nervous system by BoHV-1 or BoHV-5 were not detected. The striking differences in TLR mRNA levels during acute infection and reactivation provide evidence that the innate immune response may be involved in the clinical outcomes observed at each stage. Further research is required to analyze the mechanisms that initiate TLR activation and the signaling cascade mediated by each TLR to elucidate the precise role these receptors play in bovine herpesvirus encephalitis. Fil: Rensetti, Daniel Ernesto. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina Fil: Quintana, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentina Fil: Morán, Pedro Edgardo. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Verna, Andrea Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina Fil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina Fil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina

Objective. The aim of this study was to determine the presence of Bovine herpesvirus – 1 (BoHV- 1) and Bovine herpesvirus 5 (BoHV-5) neutralizing antibodies in herds on the Colombian high plains and their correlation with the level of cross-protection against both herpesviruses. Materials and methods . This study was carried out on cattle farms located around the towns of Puerto Lopez and Puerto Gaitan in Colombia’s Meta department. Sampling was made by convenience. Twenty-three farms were involved in the study; 488 sera samples were taken by random sampling. Virus neutralization test were performed according to the protocols of the OIE. Each serum was evaluated independently for each virus, BoHV-1 and BoHV-5. Results. The serological test confirmed the presence of BoHV-1 and BoHV-5 infections in the Colombian bovine population in 100% and 73.9% respectively. However, crossreaction for both viruses was not evident in all farms evaluated. Conclusions. Alpha-herpesviruses are amongst the most significant infectious agents affecting cattle. Bovine herpesvirus 1 (BoHV-1) is found throughout the whole world and is endemic in Colombian bovine population, whereas bovine herpesvirus 5 (BoHV-5) has limited geographical distribution, mainly being reported in South-America (Brazil and Argentina), and we also confirmed the presence of BoHV-5 in Colombia. Objetivo. El objetivo de este estudio fue determinar la presencia de anticuerpos para herpesvirus bovino 1 (BoHV-1) y herpesvirus bovino 5 (BoHV-5), asi como tambien determinar el nivel de proteccion cruzada contra los dos herpesvirus en hatos de la altillanura colombiana. Materiales y metodos . Este estudio se realizo en fincas ubicadas en los municipios de Puerto Lopez y Puerto Gaitan departamento del Meta. Se realizo un muestreo por conveniencia. Se emplearon 23 fincas, donde se realizo un muestreo al azar en cada una para un total de 488 muestras. Las muestras de suero fueron analizadas mediante sero-neutralizacion viral empleando los protocolos de la OIE. Cada uno de los sueros fue evaluado de manera independiente para cada uno de los virus BoHV-1 y BoHV-5. Resultados . Mediante la sero-neutralizacion viral se confirmo la presencia de BoHV-1 y BoHV-5 en la poblacion bovina colombiana en 100% y 73.9%, respectivamente. Sin embargo, la reactividad 2016 cruzada no fue evidente en todas las fincas evaluadas. Conclusiones . Los alfa-herpesvirus bovinos se encuentran entre los agentes infecciosos mas importantes del ganado bovino. Particularmente, el herpesvirus bovino 1 (BoHV-1) presenta una distribucion mundial, siendo endemico en la poblacion bovina colombiana, mientras que el herpesvirus bovino 5 (BoHV-5), presenta una distribucion geografica limitada; reportandose principalmente en Suramerica (Brasil y Argentina), comprobandose la presencia de este en los hatos colombianos.