Presence of co-infection between bovine leukemia virus and bovine herpesvirus 1 in herds vaccinated against bovine respiratory complex.

The aim of this study was molecular identification of bovine leukemia virus and possible co-infection with bovine respiratory disease complex (BRDC) viral agents in Mexican dairy herds. We collected 533 blood samples from cattle vaccinated against the BRDC virus in 9 states across Mexico. Peripheral blood leukocytes were removed and genetic material was extracted to detect bovine leukemia virus (BLV), bovine herpesvirus 1 (BoHV-1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV-3), and bovine respiratory syncytial virus (BRSV) infection using polymerase chain reaction. We identified high BLV infection rates in 270 cattle (50.65%). One hundred and thirty-three cows (24.95%) tested positive for BoHV-1, of which 65 samples were positive for both viruses (BoHV-1 and BLV) and 68 were only positive for BoHV-1. Only 4 samples tested positive for BPIV-3 and no sample was positive for BVDV or BRSV. Relative risk and odds ratio analyses did not identify that the presence of BLV infection favors BoHV-1 co-infection in vaccinated herds.

Le but de cette étude était l'identification moléculaire du virus de la leucémie bovine et une éventuelle co-infection par des agents viraux du complexe des maladies respiratoires bovines (BRDC) dans des troupeaux laitiers mexicains. Nous avons recueilli 533 échantillons de sang de bovins vaccinés contre le virus BRDC dans neuf états du Mexique. Les leucocytes du sang périphérique ont été prélevés et le matériel génétique a été extrait pour détecter le virus de la leucémie bovine (BLV), le virus de l'herpès bovin 1 (BoHV-1), le virus de la diarrhée virale bovine (BVDV), le virus parainfluenza bovin 3 (BPIV-3), et le virus respiratoire syncytial bovin (BRSV) par réaction d'amplification en chaîne par la polymérase. Nous avons identifié des taux élevés d'infection par le BLV chez 270 bovins (50,65 %). Cent trente-trois bovins (24,95 %) ont été testés positifs pour le BoHV-1, desquels 65 échantillons étaient positifs pour les deux virus (BoHV-1 et BLV) et 68 étaient uniquement positifs pour le BoHV-1. Seuls quatre échantillons ont été testés positifs pour le BPIV-3 et aucun échantillon n'a été positif pour le BVDV ou le BRSV. Les analyses du risque relatif et des rapports de cotes n'ont pas identifié que la présence d'une infection par le BLV favorise la co-infection par le BoHV-1 dans les troupeaux vaccinés.(Traduit par les auteurs).

Serologic profile of bovine Alfaherpesvirus type1 (BoHV-1) and Pestivírus A (BVDV-1) in herds of a family farming of the Sao Paulo state / Perfil sorológico do Alfaherpesvirus bovino tipo 1 (BoHV-1) e Pestivírus A (BVDV-1) em rebanhos da agricultura familiar no estado de São Paulo

Serum samples from 719 dairy cattle of reproductive age, from 47 farms in a settlement in the municipality of Andradina, São Paulo, were analyzed for the presence of antibodies against the Nebraska strain of BoHV-1 and Singer type 1 of BVDV-1, using the virus neutralization technique (VN). Regarding BoHV-1, 72.0% of the samples analyzed (518/719) were positive, with a geometric mean of antibody titers of 318. For BVDV-1, 44.8% of the samples were reactive, only one farm had no reactive animals, and the majority of antibody titers found in the animals' sera were low. The results of this study revealed that BoHV-1 was present in all of the studied herds, with moderate titration in the majority of cases, with the presence of some high individual titers among the tested animals. Also, even with the reactivation or circulation of BoHV-1 in herds, the clinical disease was not observed, an epidemiological finding that also applied to BVDV-1. In view of the results obtained, the circulation of diseases in family farm herds in the studied region was evident, suggesting the need to study local risk factors and improve health education policies as a way to prevent or reduce the prevalence of these diseases.(AU)

Coletou-se 719 amostras de soro sanguíneo de bovinos em idade reprodutiva, de 47 propriedades de um assentamento no município de Andradina-SP. As amostras foram analisadas para presença de anticorpos contra ambos os vírus estudados, pela técnica de virusneutralização, utilizando-se a estirpe Nebraska para o BoHV-1 e Singer tipo 1 para o BVDV-1. Em relação ao BoHV-1, 72,0% (518/719) das amostras foram positivas, sendo 318 a média geométrica dos títulos de anticorpos (GMT).Para o BVDV-1, 44,8% das amostras foram reagentes; apenas uma propriedade não teve animais reagentes, sendo a maioria dos títulos de anticorpos encontrados nos soros baixa. Observou-se que o BoHV-1 está presente em todos os rebanhos estudados com titulação moderada, com presença de alguns títulos individuais altos entre animais e com GMT alta em um dos rebanhos. Mesmo havendo a reativação ou mesmo a circulação do BoHV-1 nos rebanhos, não foi observado a doença clínica, fato epidemiológico que também ocorreu com o BVDV-1. Diante dos resultados obtidos, ficou evidente a circulação das doenças nos rebanhos da agricultura familiar na região estudada, sugerindo o estudo de fatores de risco local e melhorias de políticas de educação em saúde como forma de prevenir e ou reduzir a prevalência das enfermidades.(AU)

Serologic profile of bovine Alfaherpesvirus type1 (BoHV-1) and Pestivírus A (BVDV-1) in herds of a family farming of the Sao Paulo state / Perfil sorológico do Alfaherpesvirus bovino tipo 1 (BoHV-1) e Pestivírus A (BVDV-1) em rebanhos da agricultura familiar no estado de São Paulo

Serum samples from 719 dairy cattle of reproductive age, from 47 farms in a settlement in the municipality of Andradina, São Paulo, were analyzed for the presence of antibodies against the Nebraska strain of BoHV-1 and Singer type 1 of BVDV-1, using the virus neutralization technique (VN). Regarding BoHV-1, 72.0% of the samples analyzed (518/719) were positive, with a geometric mean of antibody titers of 318. For BVDV-1, 44.8% of the samples were reactive, only one farm had no reactive animals, and the majority of antibody titers found in the animals' sera were low. The results of this study revealed that BoHV-1 was present in all of the studied herds, with moderate titration in the majority of cases, with the presence of some high individual titers among the tested animals. Also, even with the reactivation or circulation of BoHV-1 in herds, the clinical disease was not observed, an epidemiological finding that also applied to BVDV-1. In view of the results obtained, the circulation of diseases in family farm herds in the studied region was evident, suggesting the need to study local risk factors and improve health education policies as a way to prevent or reduce the prevalence of these diseases.

Coletou-se 719 amostras de soro sanguíneo de bovinos em idade reprodutiva, de 47 propriedades de um assentamento no município de Andradina-SP. As amostras foram analisadas para presença de anticorpos contra ambos os vírus estudados, pela técnica de virusneutralização, utilizando-se a estirpe Nebraska para o BoHV-1 e Singer tipo 1 para o BVDV-1. Em relação ao BoHV-1, 72,0% (518/719) das amostras foram positivas, sendo 318 a média geométrica dos títulos de anticorpos (GMT).Para o BVDV-1, 44,8% das amostras foram reagentes; apenas uma propriedade não teve animais reagentes, sendo a maioria dos títulos de anticorpos encontrados nos soros baixa. Observou-se que o BoHV-1 está presente em todos os rebanhos estudados com titulação moderada, com presença de alguns títulos individuais altos entre animais e com GMT alta em um dos rebanhos. Mesmo havendo a reativação ou mesmo a circulação do BoHV-1 nos rebanhos, não foi observado a doença clínica, fato epidemiológico que também ocorreu com o BVDV-1. Diante dos resultados obtidos, ficou evidente a circulação das doenças nos rebanhos da agricultura familiar na região estudada, sugerindo o estudo de fatores de risco local e melhorias de políticas de educação em saúde como forma de prevenir e ou reduzir a prevalência das enfermidades.

Mycoplasma bovis and viral agents associated with the development of bovine respiratory disease in adult dairy cows.

The etiology and pathologic findings of bovine respiratory disease (BRD) in adult dairy cows (n = 35) from a commercial dairy herd in Southern Brazil were investigated. Pulmonary samples were examined for histopathologic patterns and specific features within these patterns, while immunohistochemical (IHC) assays were designed to detect the intralesional antigens of viral infectious disease agents and Mycoplasma bovis. Pneumonia was diagnosed in 91.4% (32/35) of these cases; neither pneumonia nor any of the infectious disease pathogens evaluated occurred in three cows. The presence of multiple respiratory pathogens in 75% (24/32) of these cases indicated the complex origin of pneumonia in cattle. Interstitial pneumonia, necrosuppurative bronchopneumonia and suppurative bronchopneumonia were the principal patterns of pulmonary disease identified by histopathology. The most frequent pathogens identified by IHC were bovine viral diarrhea virus (BVDV; n = 18), M. bovis (n = 16) and bovine alphaherpesvirus type 1 (BoHV-1; n = 14), followed by bovine respiratory syncytial virus (BRSV; n = 11) and bovine parainfluenza virus type 3 (BPIV-3; n = 5). Obliterative bronchiolitis and peribronchial lymphocytic cuffings were the characteristic histopathologic features associated with M. bovis. Necrohemorrhagic bronchitis with bronchial angiogenesis was associated with BoHV-1. Necrotizing bronchitis and bronchiolitis were associated with BVDV, BoHV-1 and BRSV. Ballooning degeneration of the bronchial and bronchiolar epithelia was associated with BRSV and BoHV-1. This is the first report from Brazil that correlated the histopathologic findings of BRD with the associated infectious disease agents by immunohistochemistry. M. bovis was frequently detected in the tissues of cows with fatal pulmonary disease during this study and may be a possible primary disease pathogen associated with the development of BRD in dairy cows. Additionally, the histopathologic features identified within patterns of pulmonary disease during this investigation may be an efficient diagnostic tool to associate histopathologic findings with specific agents of BRD in dairy cows.

A molecular survey using a validated real-time PCR assay finds no evidence of bovine alphaherpesvirus 2 in samples from animals with suspected vesicular disease in Brazil between 2014 and 2017.

Bovine alphaherpesvirus 2 (BoHV-2) is the etiologic agent of bovine mammillitis (BM) and pseudo-lumpy skin disease. BM is also important because its clinical presentation can be confused with foot-and-mouth disease (FMD), making it necessary to establish differential diagnoses and perform additional laboratory tests. The objective of this work was to use a validated real-time PCR assay to test for the presence of BoHV-2 in samples from cattle and buffalo with suspected vesicular disease in Brazil. The method could detect the virus at a concentration of 0.5 fg/µL and had 99.4% amplification efficiency, a repeatability error of only 4.1%, and good reproducibility with other reagents. No evidence of BoHV-2 causing vesicular disease in cattle and buffalo was found in this work. This study was able to validate a new methodology for detection of BoHV-2 and evaluate its usefulness for investigating outbreaks of vesicular disease Brazil. The importance of BoHV-2 in cases involving other clinical signs should still be studied using the qPCR developed in this work.

Bayesian estimation of herd-level prevalence and risk factors associated with BoHV-1 infection in cattle herds in the State of Paraíba, Brazil.

A cross-sectional study was carried out to estimate the animal- and herd-level prevalence of bovine herpesvirus 1 (BoHV-1) infection in cattle in the State of Paraíba, and to identify risk factors associated with herd-level infection. The state was divided into three sampling strata, and for each stratum, the prevalence of herds infected with BoHV-1 was estimated through a two-stage sampling survey carried out from September 2012 to January 2013. In total, 2443 animals were sampled from 478 herds. A virus-neutralization test was used for BoHV-1 antibody detection. A Bayesian latent-class model was used to describe the data, taking into account imperfect diagnostic test characteristics and the non-independence of test results from animals within the same herd, and using a dynamic within-model risk factor selection method based on indicator variable selection. The adjusted herd-level prevalence was estimated to be 84% (95% CI: 80-88%) for the State of Paraíba, and the animal-level prevalence was estimated to be 73% (95% CI: 66-84%). Only five of the available risk factors were used by the model, with the three most influential being disposal of aborted foetuses (3.78, 95% CI: 1.11-13.85), sharing resources with other farms (3.0, 95% CI: 1.1-8,6), and a herd size of > 23 animals (2.5, 95% CI: 1.1-6.0). Our findings suggest that the animal- and herd-level seroprevalence of BoHV-1 infection in the State of Paraíba is high. While some risk factors such as herd size and sharing resources were identified as risk factors for BoHV-1 infection, these risk factors are initially likely to be of only minor relevance in a control programme due to the extremely high prevalence of infected farms. However, the results are relevant to the risk of reintroduction of disease on farms that have previously eradicated the disease.

Latent bovine herpesvirus 1 and 5 in milk from naturally infected dairy cattle.

Bovine herpesvirus 1 and 5 (BoHV-1 and -5) are antigenically and genetically related and can establish latent infection. We aimed to analyze the applicability of the milk sample to detect latently BoHV-infected cattle. BoHV-1 non-vaccinated clinically healthy cows from five dairy cattle herds (herd 1, n=24; herd 2, n=39; herd 3, n=39; herd 4, n=36; herd 5, n=70) were studied. We confirmed the presence of BoHV-1, and for the first time, BoHV-5 in the milk of naturally infected dairy cattle.

Development of a nanogold slot blot inhibition assay for the detection of antibodies against bovine herpesvirus type 1.

Bovine herpesvirus type 1 (BoHV-1) is recognized as an important pathogen causing respiratory, reproductive, and neurological disorders in cattle and is associated with economic losses to animal industry. Accurate diagnostic methods are needed for prevention of disease transmission. While the virus neutralization test is considered the gold standard method, it requires maintenance of the virus and cell cultures, which is time consuming and expensive. Serological techniques such as enzyme-linked immunosorbent assay (ELISA) are widely applied, as these are easy to perform and provide quick results. In the present study, a nanogold slot blot inhibition assay was developed for the serological diagnosis of BoHV-1 and compared with standard ELISA and horseradish peroxidase (HRP) slot blot assays. Of 42 serum samples tested by ELISA, 32 (76.2%) were positive and 10 (23.8%), were negative. The sensitivity and specificity of the nanogold slot blot inhibition assay was similar to that observed for ELISA and HRP slot blot assays, and a strong correlation was observed between the tests. Thus, the nanogold slot blot inhibition assay may serve as an efficient and rapid alternative to ELISA in settings, where plate-reading equipment is lacking.

Molecular survey of infectious agents associated with bovine respiratory disease in a beef cattle feedlot in southern Brazil.

We investigated the occurrence of infectious pathogens during an outbreak of bovine respiratory disease (BRD) in a beef cattle feedlot in southern Brazil that has a high risk of developing BRD. Nasopharyngeal swabs were randomly collected from steers ( n = 23) and assessed for the presence of infectious agents of BRD by PCR and/or RT-PCR assays. These included: Histophilus somni, Mannheimia haemolytica, Pasteurella multocida, Mycoplasma bovis, bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCoV), bovine viral diarrhea virus (BVDV), bovine alphaherpesvirus 1 (BoHV-1), and bovine parainfluenza virus 3 (BPIV-3). Pulmonary sections of one steer that died with clinical BRD were submitted for pathology and molecular testing. The frequencies of the pathogens identified from the nasopharyngeal swabs were: H. somni 39% (9 of 23), BRSV 35% (8 of 23), BCoV 22% (5 of 23), and M. haemolytica 13% (3 of 23). PCR or RT-PCR assays did not identify P. multocida, M. bovis, BoHV-1, BVDV, or BPIV-3 from the nasopharyngeal swabs. Single and concomitant associations of infectious agents of BRD were identified. Fibrinous bronchopneumonia was diagnosed in one steer that died; samples were positive for H. somni and M. haemolytica by PCR. H. somni, BRSV, and BCoV are important disease pathogens of BRD in feedlot cattle in Brazil, but H. somni and BCoV are probably under-reported.

Involvement of toll-like receptors 3 and 7/8 in the neuropathogenesis of bovine herpesvirus types 1 and 5.

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.

Detection of multiple viral infections in cattle and buffalo with suspected vesicular disease in Brazil.

Vesicular diseases are of high importance for livestock, primarily because of foot-and-mouth disease (FMD), which is a high-morbidity disease that generates direct losses caused by low milk production, weight loss, and indirect losses because of the need for sanitary barriers. Other vesicular diseases are also of importance for livestock because of direct impacts or because their clinical signs may be confused with those of FMD. We report herein the detection of multiple infections in cattle with suspected vesicular disease in the Brazilian states of Amazonas (AM), Mato Grosso (MT), and Roraima. Thirty-seven epithelial samples from cattle and 1 sample from a buffalo were sent to the laboratory for testing for FMDV and similar disease agents. All samples from MT were positive for parapoxvirus (Pseudocowpox virus and Bovine papular stomatitis virus). In addition, 3 samples were positive for Bluetongue virus, and 5 samples were positive for Bovine herpesvirus 1 Among these samples, 1 was positive for all of these 3 agents. Only 2 samples from AM were negative for parapoxvirus. The molecular tests conducted in this study detected multiple infections, with a high prevalence of parapoxvirus.

Distribution of bovine herpesvirus type 1 in the nervous system of experimentally infected calves.

Bovine herpesvirus type 1 (BoHV-1) is responsible for respiratory and genital disease in cattle. BoHV-1 encephalitis is only occasionally reported. However, several cases of neurological disease have been recently attributed to BoHV-1. In this study, the distribution and pathological alterations caused by two BoHV-1 strains in the nervous system of experimentally infected calves during acute infection and reactivation are described. Calves were inoculated intranasally with BoHV-1 Los Angeles (BoHV-1.LA) or Cooper (BoHV-1.Cooper) strains. Acutely infected calves were euthanased at 6 days (BoHV-1.Cooper, n = 2) and 7 days post-inoculation (BoHV-1.LA, n = 2). Latently infected calves that were given dexamethasone to induce reactivation were euthanased at 2 days (BoHV-1.Cooper, n = 2) or 5 days (BoHV-1.LA, n = 2) after dexamethasone administration. Both BoHV-1 strains were isolated from the brains of acutely infected calves. Distribution of viral DNA in the neural tissues was similar for both strains. During reactivation, neither BoHV-1.LA nor BoHV-1.Cooper was isolated from any brain section or trigeminal ganglia in infected calves. Macroscopic lesions were not evident in any group. In BoHV-1.LA infected calves, microscopic lesions were found in the brain but not in the trigeminal ganglia. Microscopic lesions in the brain of BoHV-1.Cooper infected calves were not as evident as in BoHV-1.LA infected animals. However, mononuclear infiltrates and neuronophagia were present in trigeminal ganglia. The results of this study demonstrated that respiratory BoHV-1 strains are able to replicate and disseminate within the bovine nervous tissue and provide evidence of the neuroinvasiveness of BoHV-1 strains.

A new method for simultaneous detection and discrimination of Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) using real time PCR with high resolution melting (HRM) analysis.

Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are antigenically and genetically similar. The aim of this study was to develop a simple and reliable one-step real time PCR assay with high resolution melting (HRM) analysis for the simultaneous detection and differentiation of BoHV-1 and BoHV-5. Optimization of assay conditions was performed with DNA from reference strains. Then, DNA from field isolates, clinical samples and tissue samples of experimentally infected animals were studied by real time PCR-HRM. An efficient amplification of real time PCR products was obtained, and a clear melting curve and appropriate melting peaks for both viruses were achieved in the HRM curve analysis for BoHV type identification. BoHV was identified in all of the isolates and clinical samples, and BoHV types were properly differentiated. Furthermore, viral DNA was detected in 12/18 and 7/18 samples from BoHV-1- and BoHV-5-infected calves, respectively. Real time PCR-HRM achieved a higher sensitivity compared with virus isolation or conventional PCR. In this study, HRM was used as a novel procedure. This method provides rapid, sensitive, specific and simultaneous detection of bovine alpha-herpesviruses DNA. Thus, this technique is an excellent tool for diagnosis, research and epidemiological studies of these viruses in cattle.

Frequency of antibodies against bovine herpesvirus type 1 (BoHV-1) in beef cattle not vaccinated / Frequência de anticorpos contra o herpesvírus bovino tipo 1 (BoHV-1) em bovinos de corte não vacinados

Bovine herpesvirus type 1 (BoHV-1), is responsible for clinical manifestations such as infectious bovine rhinotracheitis, abortion, conjunctivitis, infectious pustular vulvovaginitis and balanoposthitis. This virus has been responsible for major losses in different productive and reproductive herds in the country. Thus, the objective of this study was to estimate the frequency of antibodies against BoHV-1 in beef heifers not vaccinated in Microregion of Imperatriz, Maranhao, and identify the age group most affected by the virus, as well as a study of factors associated with virus infection and to evaluate the indirect ELISA using the serum neutralization (SN) as a reference standard. The study was conducted in 48 herds, cutting, distributed in 12 counties of Microregion of Imperatriz. The samples were collected from female cattle stratified into three age groups, ? 12 months, between 12 and 36 months and ? 36 months of age. The samples were subjected to two serological tests, ELISA and SN. In each herd, an epidemiological questionnaire was applied in order to obtain information on management and reproductive sanitary, for the study of risk factors. The frequency of antibodies against BoHV-1 in Microregion of Imperatriz was 63.23%, and the municipalities of Açailândia Buritirana showed the highest frequencies, both with 80.44%, the most affected age group, the Microregion, was animals aged ≥ 36 months (69.65%). Based on the results we can conclude that the frequency of antibodies against BoHV-1 is high, between the age groups most affected were the animals aged ≥ 36 months were considered risk factors for virus transmission, return to estrus (OR=1.874), recovery of animals from other states / region (OR=1.365) and the creation of goat / sheep associated with bovine (OR=1.348), the indirect ELISA technique showed moderate concordance when compared to SN technique, which is the gold standard technique for diagnosis of BoHV-1.(AU)

O herpesvírus bovino tipo 1 (BoHV-1), é responsável por manifestações clínicas como a rinotraqueíte infecciosa bovina, abortamentos, conjuntivite, balanopostite e vulvovaginite pustular infecciosa. Esse vírus tem sido responsável por grandes prejuízos produtivos e reprodutivos em diversos rebanhos do país. Deste modo, o objetivo desse trabalho foi estimar a frequência de anticorpos contra o BoHV-1 em fêmeas bovinas de corte não vacinadas, identificar a faixa etária mais acometida, bem como, realizar um estudo dos fatores associados à infecção do vírus e avaliar a técnica ELISA indireto utilizando a soroneutralização (SN) como padrão de referência. O estudo foi realizado em 48 rebanhos de corte, distribuídos em 12 municípios da Microrregião de Imperatriz, Maranhão. As amostras foram coletadas de fêmeas bovinas estratificadas em três faixas etárias, ? 12 meses, entre 12 e 36 meses e ? 36 meses de idade. Os testes empregados foram: ELISA indireto e SN. Em cada rebanho, foi aplicado um questionário epidemiológico, com o objetivo de obter informações sobre manejo sanitário e reprodutivo, para o estudo de fatores de risco. A frequência de anticorpos contra o BoHV-1 na Microrregião de Imperatriz foi de 63,23% (n=698), os municípios de Açailândia e Buritirana, apresentaram maiores frequências, ambos com 80,44% (n=74). a faixa etária mais acometida, foram a dos animais com idade ≥ 36 meses; foram considerados fatores de risco para a transmissão do vírus, retorno ao cio (OR=1,874), reposição de animais oriundos de outros estados/região (OR=1,365) e a criação de caprinos/ovinos associados com bovinos (OR=1,348); a técnica ELISA indireta apresentou concordância moderada quando comparada a técnica de SN, que é a técnica padrão ouro de diagnóstico para BoHV-1.(AU)

Relevamiento serológico de anticuerpos contra enfermedades virales de interés sanitario en llamas (Lama glama) de la provincia de Jujuy, Argentina / Serological survey of antibodies aainst vital diseases of public health interest in llamas (Lama glama) from Jujuy rpovince, Argentina

Las poblaciones de llamas de Argentina se concentran principalmente en la provincia de Jujuy; su explotación representa un importante recurso económico de las comunidades altoandinas. El objetivo de este trabajo fue evaluar la seroprevalencia de anticuerpos contra algunos agentes virales asociados a enfermedades de impacto productivo en rodeos de llamas de Jujuy. Se analizaron 349 sueros de llamas adultas de 6 departamentos de la puna jujeña ubicados por encima de los 3300 msnm. Se obtuvo una prevalencia del 100 % para rotavirus grupo A y del 70 % para el virus parainfluenza-3 bovino, mientras que no se detectaron reactores para herpesvirus bovino 1, virus de la diarrea viral bovina, influenza A humana (H1N1) e influenza equina (H3N8). Los resultados obtenidos confirman la amplia distribución de rotavirus y virus parainfluenza y la baja susceptibilidad a herpesvirus y pestivirus en las tropas de llamas de la puna jujeña

Llama population from Argentina is mainly concentrated in the Andean Puna, Jujuy. Llamas represent an important economic resource for the Andean communities. The aim of this study was to investigate the prevalence of antibodies against viral antigens associated to viral diseases of economic impact (neonatal diarrhea, reproductive and respiratory syndromes). A total of 349 serum samples from adult llamas were analyzed. The obtained antibody prevalence was 100 % for Rotavirus A and 70 % for Bovine parainfluenza virus 3. In contrast, no reactors were detected to Bovine herpesvirus 1, Bovine viral diarrhea virus 1, Human influenza A virus (H1N1) and Equine influenza virus (H3N8). These results confirm the wide circulation of rotavirus and parainfluenza virus in Argentinean llamas and suggest that susceptibility to infection with bovine herpesvirus, pestivirus and influenza A viruses is low. This serologic survey provides novel information regarding the epidemiology of viral diseases affecting llamas from the Argentinean Andean Puna

Frequency of antibodies against bovine herpesvirus type 1 (BoHV-1) in beef cattle not vaccinated / Frequência de anticorpos contra o herpesvírus bovino tipo 1 (BoHV-1) em bovinos de corte não vacinados

Bovine herpesvirus type 1 (BoHV-1), is responsible for clinical manifestations such as infectious bovine rhinotracheitis, abortion, conjunctivitis, infectious pustular vulvovaginitis and balanoposthitis. This virus has been responsible for major losses in different productive and reproductive herds in the country. Thus, the objective of this study was to estimate the frequency of antibodies against BoHV-1 in beef heifers not vaccinated in Microregion of Imperatriz, Maranhao, and identify the age group most affected by the virus, as well as a study of factors associated with virus infection and to evaluate the indirect ELISA using the serum neutralization (SN) as a reference standard. The study was conducted in 48 herds, cutting, distributed in 12 counties of Microregion of Imperatriz. The samples were collected from female cattle stratified into three age groups, ? 12 months, between 12 and 36 months and ? 36 months of age. The samples were subjected to two serological tests, ELISA and SN. In each herd, an epidemiological questionnaire was applied in order to obtain information on management and reproductive sanitary, for the study of risk factors. The frequency of antibodies against BoHV-1 in Microregion of Imperatriz was 63.23%, and the municipalities of Açailândia Buritirana showed the highest frequencies, both with 80.44%, the most affected age group, the Microregion, was animals aged ≥ 36 months (69.65%). Based on the results we can conclude that the frequency of antibodies against BoHV-1 is high, between the age groups most affected were the animals aged ≥ 36 months were considered risk factors for virus transmission, return to estrus (OR=1.874), recovery of animals from other states / region (OR=1.365) and the creation of goat / sheep associated with bovine (OR=1.348), the indirect ELISA technique showed moderate concordance when compared to SN technique, which is the gold standard technique for diagnosis of BoHV-1.

O herpesvírus bovino tipo 1 (BoHV-1), é responsável por manifestações clínicas como a rinotraqueíte infecciosa bovina, abortamentos, conjuntivite, balanopostite e vulvovaginite pustular infecciosa. Esse vírus tem sido responsável por grandes prejuízos produtivos e reprodutivos em diversos rebanhos do país. Deste modo, o objetivo desse trabalho foi estimar a frequência de anticorpos contra o BoHV-1 em fêmeas bovinas de corte não vacinadas, identificar a faixa etária mais acometida, bem como, realizar um estudo dos fatores associados à infecção do vírus e avaliar a técnica ELISA indireto utilizando a soroneutralização (SN) como padrão de referência. O estudo foi realizado em 48 rebanhos de corte, distribuídos em 12 municípios da Microrregião de Imperatriz, Maranhão. As amostras foram coletadas de fêmeas bovinas estratificadas em três faixas etárias, ? 12 meses, entre 12 e 36 meses e ? 36 meses de idade. Os testes empregados foram: ELISA indireto e SN. Em cada rebanho, foi aplicado um questionário epidemiológico, com o objetivo de obter informações sobre manejo sanitário e reprodutivo, para o estudo de fatores de risco. A frequência de anticorpos contra o BoHV-1 na Microrregião de Imperatriz foi de 63,23% (n=698), os municípios de Açailândia e Buritirana, apresentaram maiores frequências, ambos com 80,44% (n=74). a faixa etária mais acometida, foram a dos animais com idade ≥ 36 meses; foram considerados fatores de risco para a transmissão do vírus, retorno ao cio (OR=1,874), reposição de animais oriundos de outros estados/região (OR=1,365) e a criação de caprinos/ovinos associados com bovinos (OR=1,348); a técnica ELISA indireta apresentou concordância moderada quando comparada a técnica de SN, que é a técnica padrão ouro de diagnóstico para BoHV-1.

Evidence of bovine viral diarrhea, but absence of infectious bovine rhinotracheitis and bovine brucellosis in the endangered huemul deer (Hippocamelus bisulcus) in Chilean Patagonia.

We screened 18 endangered Chilean huemul (Hippocamelus bisulcus) for antibodies to infectious agents. We detected no antibody to bovine herpesvirus-1 (BHV-1) or Brucella abortus (BA); two huemul had antibody to bovine viral diarrhea virus (BVDV). Cattle (n=35) had antibody to BVDV and BHV-1 but not BA.

Serological evidence and risk factors associated with Caprine herpesvirus 1 in dairy goat flocks in a semiarid region of northeastern Brazil.

A cross-sectional study was carried out to determine the flock-level seroprevalence of Caprine herpesvirus 1 (CpHV-1) and Bovine herpesvirus 1 (BoHV-1) and 2 (BoHV-2) and risk factors associated with CpHV-1 in dairy goat flocks from a semiarid region of northeastern Brazil. A total of 1,034 serum samples from 110 flocks were collected from March 2009 through March 2010. A structured questionnaire focusing on variables related to risk factors for CpHV-1 infection was given to each farmer at the time of blood collection. Antibodies against CpHV-1, BoHV-1, and BoHV-2 were detected by neutralization tests. The flock-level prevalences of CpHV-1, BoHV-1, and BoHV-2 were 89.1% (98/110; 95% confidence interval [CI]: 81.7-94.2), 80% (88/110; 95% CI: 71.3-87), and 4.5% (5/110; 95% CI: 1.5-10.3), respectively. Frequencies of seropositive animals were 36.6% (379/1,034), 25.8% (267/1,034), and 0.6% (6/1,034) for CpHV-1, BoHV-1, and BoHV-2, respectively. The use of natural mating was identified as a risk factor associated with CpHV-1 flock-level prevalence (P = 0.001). It is suggested that adoption of veterinary services and active surveillance of the at-risk flocks in the study region should be initiated to reduce the prevalence of herpesvirus infections.

Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and its use in an ELISA for gE antibodies / Expressão procariota de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 e uso em ELISA para anticorpos contra a gE

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.

Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and its use in an ELISA for gE antibodies / Expressão procariota de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 e uso em ELISA para anticorpos contra a gE

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.(AU)

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.(AU)