Serological Profile for Major Respiratory Viruses in Unvaccinated Cows from High-Yielding Dairy Herds.

This study aims to determine the serological profile of high-yielding dairy cows for four main viruses (bovine alphaherpesvirus 1 (BoAHV1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV3), and bovine respiratory syncytial virus (BRSV)) related to bovine respiratory disease (BRD) in cattle herds worldwide. In this survey, 497 blood serum samples were collected from non-vaccinated dairy cows without clinical respiratory signs in 39 herds in the central-eastern mesoregion of Paraná State, South Brazil. The presence of neutralizing antibodies was determined by virus neutralization (VN) tests. VN antibodies against BoAHV1, BVDV, BPIV3, and BRSV were detected in 355 (71.4%), 280 (56.3%), 481 (96.8%), and 315 (63.4%) serum samples, respectively. The frequencies of seropositive herds for BoAHV1, BVDV, BPIV3, and BRSV were 79.5 (n = 31), 82.0 (n = 32), 100 (n = 39), and 84.6% (n = 33), respectively. The frequencies of seropositive cows varied according to the type of herd management and the number of cows in the herd. The detection of VN antibodies in unvaccinated dairy cattle herds demonstrated the endemic circulation of the four viruses in the herds evaluated. For BRD prevention, it is recommended to implement a vaccination program for cows that provides passive immunity in calves and active immunity in cows.

Coding-complete genome sequences of two bovine viral diarrhea virus 1a isolates from Uruguay.

Bovine viral diarrhea virus (BVDV) is an economically relevant pathogen affecting cattle production and reproduction worldwide. We report the coding-complete sequences of two BVDV-1a subtype isolates, circulating in Uruguay.

First report of bovine viral diarrhea virus subgenotypes 1d and 1e in southern Chile.

Bovine viral diarrhea virus (BVDV) affects cattle worldwide causing severe productive and economic loss. In this study, we investigated the subgenotypes of BVDV circulating in cattle samples from the Aysén region, an active cattle breeding area located in southern Chile. Partial amplification of the 5' untranslated region (UTR) was performed by polymerase chain reaction (PCR), and twelve samples were analyzed by Sanger sequencing and phylogenetic analysis. Eight samples were identified as belonging to Pestivirus bovis subgenotype 1e, three to 1-b, and one to 1-d. The phylogenetic analyses performed revealed a marked distance between these now-identified strains and those previously reported in the country. These findings support the need to continually expand the analysis of the variability of the viral phylogeny for the currently circulating BVDV strains and to update the vaccines recommended for this livestock area and surrounding areas.

Serological profile of respiratory viruses in unvaccinated steers upon their arrival at Brazilian feedlot facilities.

Bovine viral diarrhea virus (BVDV), bovine alphaherpesvirus 1 (BoAHV1), bovine respiratory syncytial virus (BRSV), and bovine parainfluenza virus 3 (BPIV-3) are involved in bovine respiratory disease. These viruses can infect the respiratory system and cause considerable economic losses to beef and dairy cattle herds. This study aimed to determine the serological profiles of steers for BVDV, BoAHV1, BRSV, and BPIV-3 upon their arrival at Brazilian feedlot facilities. A total of 1,282 serum samples from unvaccinated steers were obtained on the first day of feeding. Samples were collected from 31 beef cattle herds reared in an extensive rearing system in six Brazilian states. Antibodies against BVDV, BoAHV1, BRSV, and BPIV-3 were detected using a virus neutralization test. The steers were distributed in agreement with their age and the Brazilian state of origin. The highest seropositivity was for BoAHV1 and BPIV-3 at 92.1% (1,154/1,253) and 86.6% (1,100/1,270), respectively. The seropositivity of BRSV was 77.1% (959/1,244). BVDV presented a lower rate, at slightly more than 50% (51.8%; 656/1,266). Age was a risk factor for the presence of antibodies against BVDV, BoAHV1, and BPIV-3 but not BRSV. A positive correlation was identified between BoAHV1 and BPIV-3 (P = 0.85) and between BRSV and BPIV-3 (P = 0.47). The high rate of seropositive steers for these four respiratory viruses on the first day of confinement identified in this serological survey provides important epidemiological information on respiratory infections, as the seropositivity of the four main bovine respiratory viruses in Brazilian beef cattle herds in an extensive rearing system.

Immune protection induced by E2 recombinant glycoprotein of bovine viral diarrhea virus in a murine model.

Bovine viral diarrhea virus (BVDV) is considered the most important viral pathogen in ruminants worldwide due to the broad range of clinical manifestations displayed by infected animals. Therefore, infection with BVDV leads to severe economic losses in several countries' beef and dairy industries. Vaccination prevents reproductive failure and gastrointestinal and respiratory disorders caused by BVDV infection. However, considering their limitations, conventional vaccines such as live, attenuated, and killed viruses have been applied. Hence, different studies have described subunit vaccines as an effective and safe alternative for BVDV protection. Therefore, in this study, the ectodomain of E2 (E2e) glycoprotein from NADL BVDV strain was expressed in mammalian cells and used in two vaccine formulations to evaluate immunogenicity and protection against BVDV conferred in a murine model. Formulations consisted of solo E2e glycoprotein and E2e glycoprotein emulsified in adjuvant ISA 61 VG. Five groups of 6 mice of 6-to-8-week-old were immunized thrice on days 1, 15, and 30 by intraperitoneal injection with the mentioned formulations and controls. To evaluate the conferred protection against BVDV, mice were challenged six weeks after the third immunization. In addition, the humoral immune response was evaluated after vaccination and challenge. Mice groups inoculated with solo E2e and the E2e + ISA 61 VG displayed neutralizing titers; however, the E2 antibody titers in the E2e + ISA 61 VG group were significantly higher than the mice group immunized with the solo E2e glycoprotein. In addition, immunization using E2e + ISA 61 VG prevents animals from developing severe lesions in surveyed tissues. Moreover, this group acquired protection against the BVDV challenge, evidenced by a significant reduction of positive staining for BVDV antigen in the lungs, liver, and brain between the experimental groups. Our findings demonstrated that using E2e + ISA 61 VG induces greater BVDV protection by an early humoral response and reduced histopathological lesions and BVDV antigen detection in affected organs, indicating that E2e + ISA 61 VG subunit formulation can be considered as a putative vaccine candidate against BVDV. The efficacy and safety of this vaccine candidate in cattle requires further investigation.

Impact of persistent bovine viral diarrhea virus infection on indicators of innate and adaptive immune function in holstein calves and cows / Impacto da infecção persistente pelo vírus da diarreia viral bovina sobre indicadoresde função imunológica inata e adaptativa em bezerras e vacas holandesas

ABSTRACT: We evaluated some indicators of innate and humoral immune response in persistently infected (PI) Holstein calves and cows from 1 to 36 months of age matched with controls from the same herd. The effects were cataloged by grouping animals into the following age groups: <12 months, 13 to 24 months, and 25 to 36 months of age. Blood samples were collected once from each animal to measure total serum protein, haptoglobin, and neutralizing antibodies titers induced by respiratory virus vaccination. Total serum protein (g/dL) was lowest in PI calves younger until 24 months old, while haptoglobin concentration was higher in PI cattle. The serum neutralizing titers against BVDV and BRSV were lower in all PI calves and cattle than in controls. PI cattle have a high serum concentration of haptoglobin, and its possible dysregulated innate immune response appears to impact the efficacy of their adaptative immune responses, resulting in poor vaccine responsiveness.

RESUMO: O objetivo desta pesquisa foi avaliar alguns indicadores da resposta imune inata e humoral em bezerros a vacas persistentemente infectadas, entre um a 36 meses de idade, pareados com controles oriundos de um mesmo rebanho. As variáveis respostas foram avaliadas agrupando-se os animais nos seguintes grupos etários: < 12 meses, 13 a 24 meses, 25 a 36 meses de idade. Amostras sanguíneas foram coletadas para mensurar as concentrações séricas de proteína, haptoglobina e anticorpos neutralizantes induzidos pela vacinação contra as viroses respiratórias. Os teores de proteína sérica total (g/dL) foram menores nos animais persistentemente infectados (PI) jovens até 24 meses de idade, enquanto que a concentração de haptoglobina foi maior nos animais PI mais velhos (25 a 36 meses). Os títulos de anticorpos neutralizantes contra o BVDV e BRSV foi menor nos animais PIs independentemente da idade, comparado com o grupo controle. Os valores reduzidos ou nulos de anticorpos contra as viroses respiratórias, combinado com a evidência de resposta imune inata desregulada, contribui com a susceptibilidade dos animais PIs para as infecções secundárias.

Impact of persistent bovine viral diarrhea virus infection on indicators of innate and adaptive immune function in holstein calves and cows / Impacto da infecção persistente pelo vírus da diarreia viral bovina sobre indicadores de função imunológica inata e adaptativa em bezerras e vacas holandesas

We evaluated some indicators of innate and humoral immune response in persistently infected (PI) Holstein calves and cows from 1 to 36 months of age matched with controls from the same herd. The effects were cataloged by grouping animals into the following age groups: <12 months, 13 to 24 months, and 25 to 36 months of age. Blood samples were collected once from each animal to measure total serum protein, haptoglobin, and neutralizing antibodies titers induced by respiratory virus vaccination. Total serum protein (g/dL) was lowest in PI calves younger until 24 months old, while haptoglobin concentration was higher in PI cattle. The serum neutralizing titers against BVDV and BRSV were lower in all PI calves and cattle than in controls. PI cattle have a high serum concentration of haptoglobin, and its possible dysregulated innate immune response appears to impact the efficacy of their adaptative immune responses, resulting in poor vaccine responsiveness.

O objetivo desta pesquisa foi avaliar alguns indicadores da resposta imune inata e humoral em bezerros a vacas persistentemente infectadas, entre um a 36 meses de idade, pareados com controles oriundos de um mesmo rebanho. As variáveis respostas foram avaliadas agrupando-se os animais nos seguintes grupos etários: < 12 meses, 13 a 24 meses, 25 a 36 meses de idade. Amostras sanguíneas foram coletadas para mensurar as concentrações séricas de proteína, haptoglobina e anticorpos neutralizantes induzidos pela vacinação contra as viroses respiratórias. Os teores de proteína sérica total (g/dL) foram menores nos animais persistentemente infectados (PI) jovens até 24 meses de idade, enquanto que a concentração de haptoglobina foi maior nos animais PI mais velhos (25 a 36 meses). Os títulos de anticorpos neutralizantes contra o BVDV e BRSV foi menor nos animais PIs independentemente da idade, comparado com o grupo controle. Os valores reduzidos ou nulos de anticorpos contra as viroses respiratórias, combinado com a evidência de resposta imune inata desregulada, contribui com a susceptibilidade dos animais PIs para as infecções secundárias.

Association of bovine viral diarrhea virus, bovine herpesvirus 1, and Neospora caninum with late embryonic losses in highly supplemented grazing dairy cows.

The objectives of this study were: 1- to evaluate the association of Bovine Viral Diarrhea Virus (BVDV), Bovine Herpes Virus 1 (BoHV-1), and Neospora caninum (N. caninum) with the risk for Late Embryonic Loss (LEL) in grazing dairy cows, 2- to evaluate blood progesterone concentration at the time of LEL occurrence, and 3- to describe a novel ultrasound-guided technique for conceptus sampling. We run a prospective cohort study involving 92 cows (46 LEL and 46 NLEL). An LEL cow was that having an embryo with no heartbeat, detached membranes, or floating structures, including embryo remnants detected at pregnancy check by ultrasonography (US) 28-42 days post-AI, whereas an NLEL cow was that with embryo heartbeats detectable by US at pregnancy check 28-42 d post-IA. We took two blood samples from every cow at pregnancy check by US (the day of LEL detection) and 28 d later to perform serological diagnosis of BVDV, BoHV-1, and N. caninum; and to measure blood progesterone concentration at pregnancy check (28-42 d post-AI). We also sampled the conceptus from all the LEL cows. We performed PCR to detect BVDV, BoHV-1, and N. caninum in sampled conceptuses from LEL cows. Finally, we evaluated the associations of risk factors (serological titers, seroconversion, and progesterone) with LEL odds with logistic models. The risk for LEL was associated with serological titers to BVDV (P = 0.03) and tended to be associated with seroconversion to BVDV, given that 19.6% (9/46) in LEL and 6.5% (3/46) in NLEL cows seroconverted to BVDV (P = 0.09). In addition, BVDV was detected in conceptuses from LEL cows that seroconverted to BVDV but not in LEL cows that did not seroconvert. Conversely, the risk for LEL was not associated with the titers or seroconversion to BoHV-1 and N. caninum. BoHV-1 and N. caninum were not identified in any of the conceptuses. Finally, blood progesterone concentration was similar in LEL and NLEL cows, and it was not associated with the risk for LEL (P = 0.54). In conclusion, BVDV infection is a risk factor for LEL in dairy cows.

HoBi-like pestivirus in 2 cases of fatal respiratory disease of feedlot cattle in Argentina.

HoBi-like pestivirus (HoBiPeV) is an emerging virus that has been detected in cattle and other ruminants. We diagnosed 2 cases of fatal bovine respiratory disease complex (BRDC) associated with infection with HoBiPeV in a feedlot in Argentina. The main findings in 2 steers autopsied were interstitial bronchopneumonia (case 1) and fibrinous bronchopneumonia (case 2). HoBiPeV was detected by RT-PCR in lungs of both animals and by immunohistochemistry in case 2. Phylogenetic analysis showed that both strains clustered within the "Brazilian-Italian" clade. In case 2, Mannheimia haemolytica was isolated from the lung. There is scant information about the contribution of HoBiPeV to the pathogenesis of BRDC. To our knowledge, HoBiPeV has not been reported previously in association with M. haemolytica pneumonia. Our findings further support the involvement of HoBiPeV in cases of BRDC and contribute to understanding the synergy of this etiologic agent in the pathogenesis of BRD, which is critical for the development of appropriate preventive strategies.

A Serosurvey for Ruminant Pestivirus Exposure Conducted Using Sera From Stray Mexico Origin Cattle Captured Crossing Into Southern Texas.

The US Department of Agriculture (USDA), Animal Plant Health Inspection Service (APHIS), Cattle Fever Tick Eradication Program (CFTEP) monitor a quarantine zone along the Texas border to prevent the introduction of stray livestock carrying cattle fever ticks entering the United States from Mexico. Stray cattle collected by CFTEP are checked for ticks and several infectious disease-causing pathogens, but not for bovine viral diarrhea virus (BVDV). BVDV is one of the most economically impactful viruses affecting US cattle producers. BVDV is present in all parts of the world, but it has been demonstrated that another distantly related pestivirus, HoBi-like pestivirus (HoBiPev), can also cause BVD. To date, HoBiPev has not been detected in the United States, but is commonly found in Brazil, and sporadically in Europe and Asia. The objective of the current study was to evaluate the seroprevalence of pestiviruses, with a specific focus on HoBiPev, in stray cattle. Virus neutralization (VN) assay was used to determine seroprevalence (or antibody titers) of BVDV-1, BVDV-2, and HoBiPev. Approximately 50% (67 of 134) of the samples were seropositive for pestiviruses; all 67 positive samples were positive (50%) for BVDV-1, 66 samples of the 67 were positive (49.3%) for BVDV-2, and the same 66 samples of the 67 were also positive (49.3%) for HoBiPev. Due to the antigenic cross-reactivity among Pestiviruses, the comparative antibody against each pestivirus was calculated from all VN-positive samples. Titers were clearly higher against BVDV-1, and only one sample had a titer clearly higher against BVDV-2. No sample had an antibody titer higher for HoBiPev, and while this does not prove the absence of HoBiPev, it does provide evidence that the prevalence of HoBiPev is less predominant than BVDV-1. Additionally, data from these samples provide evidence on the susceptibility of animals that may enter into the United States, with ~50% of the animals seronegative for bovine pestiviruses. This cattle population provides a unique opportunity to evaluate and monitor changes in seroprevalence of economically important cattle diseases affecting the cattle industry.

Bovine Viral Diarrhea Virus within and herd prevalence on pasture-based dairy systems, in southern Chile dairy farms.

Bovine Viral Diarrhea (BVD) is a ubiquitous endemic disease in most bovine populations that causes a negative economic impact on cattle production systems. However, true herd and individual prevalence in Chile have not been estimated based on statistical and epidemiological considerations and uncertainty regarding the infection's extension at animal and herd levels. In addition, the risk factors that can influence how the infection has spread among the cattle premises have not been assessed yet. Therefore, the study aimed to estimate true herd and animal prevalence of active BVDV infection in pasture-based dairy herds and to evaluate risk factors potentially associated with the status of herds and the within-herd prevalence, using a Bayesian approach. A cross-sectional study was performed over a random stratified (proportional to herd size) sample of one-hundred and fifty dairy herds selected in Chile's two leading dairy regions (Region de Los Ríos and Region de Los Lagos). We obtained 12.311 blood samples from lactating cows, but 4.998 randomly selected samples were processed due to budget constraints. For estimating BVDV Herd Prevalence (HP), a herd was considered infected if at least one positive animal to Ag-ELISA was found. In addition, a survey was completed on each farm, aiming to collect information to assess the influence of some variables over the state of the herds; both estimations were obtained using a Bayesian approach. True herd prevalence and true individual prevalence of BVDV active infection were estimated at 77 % and 3.5 % respectively, no significant differences were found between characteristics like herd size, location, vaccination, etc. Open herds and herds that mix herd categories are more likely to have higher BVDV prevalence.

Bovine Viral Diarrhea Virus in Cattle From Mexico: Current Status.

Bovine viral diarrhea (BVD) is an infectious disease, globally-distributed, caused by bovine Pestiviruses, endemic of cattle and other ruminant populations. BVD leads to significant economic losses to the cattle industry due to the wide range of clinical manifestations, including respiratory and gastrointestinal diseases and reproductive disorders. Within the Pestivirus genus of the family Flaviviridae three viral species are associated with BVD; Pestivirus A (Bovine viral diarrhea virus 1, BVDV-1), Pestivirus B (Bovine viral diarrhea virus 2, BVDV-2), and Pestivirus H (HoBi-like pestivirus, atypical ruminant pestivirus). These species are subdivided into subgenotypes based on phylogenetic analysis. The extensive genetic diversity of BVDV has been reported for several countries, where the incidence and genetic variation are more developed in Europe than in the Americas. The first report of BVDV in Mexico was in 1975; this study revealed seropositivity of 75% in cows with a clinical history of infertility, abortions, and respiratory disease. Other studies have demonstrated the presence of antibodies against BVDV with a seroprevalence ranging from 7.4 to 100%. Recently, endemic BVDV strains affecting cattle populations started to be analyzed, providing evidence of the BVDV diversity in several states of the country, revealing that at least four subgenotypes (BVDV-1a, 1b, 1c, and 2a) are circulating in animal populations in Mexico. Little information regarding BVD epidemiological current status in Mexico is available. This review summarizes available information regarding the prevalence and genetic diversity viruses associated with BVD in cattle from Mexico.

Seroprevalence of bovine viral diarrhea virus (BVDV) in cattle from Sotaquirá, Colombia.

Worldwide distributed Bovine Viral Diarrhea Virus (BVDV) represents a high risk of infection in most bovine farms, in which it is associated with gastrointestinal, respiratory, and reproductive diseases. The purpose of this research was to establish the seroprevalence and the main risk factors associated with the presentation of BVDV in the municipality of Sotaquirá, Colombia. Samples were taken from 1000 cattle of Holstein, Ayrshire, Jersey, Normande Gyr and Holstein x Gyr. Epidemiological surveys were implemented, reproductive and management variables were taken into consideration. Indirect ELISA was performed to detect specific antibodies against BVDV using the commercial kit SERELISA® BVD p80 Ab Mono Blocking. The overall seroprevalence of antibodies against BVDV was 42.5% (425/1000), where the Gyr breed (59.1% apparent prevalence (AP); 60.3% real prevalence (PR)) and the age group > 4 years (53.0% PA; 54.4% PR) presented the highest seroprevalences. A significant statistical association was found for the breed, age, management practices evaluated and the presentation of PI3 (p ≤ 0.05). Age group > 4 years, Normande breed, presentation of PI3 and grazing lease were established as risk factors associated with BVDV in the herds. These infections are mainly associated with dairy cattle and herds with many animals, so it is important to consider vaccination plans as a preventive system and follow up on the most common diseases.

Cattle seroprevalence and risk factors associated with bovine viral diarrhea in the northeastern of Colombia.

Bovine viral diarrhea (BVD) is one of the most common and economically significant viral infections in cattle. Different risk factors have been associated with viral exposure and transmission, generating endemic regions with different biotype circulations. To find the BVD status in the northeastern region of Colombia, this study aimed to determine the seroprevalence and risk factors associated with the disease in non-vaccinated farms. For this purpose, a two-time point sampling strategy was developed, obtaining serum from 1157 animals housed in 25 farms distributed in 46 townships, thus for antibody detection against non-structural protein 3 (NS3, p80) of bovine viral diarrhea virus (BVDV) using an indirect ELISA test. For the first time in Colombia, the presence of serological persistent cattle was evaluated in 1047 animals 3 weeks later of first sampling. The information on a standardized questionnaire with closed and dichotomic answers was used to calculate the putative risk factors. The association analysis by univariate and multivariate logistic regression reported odds ratios (OR) with a 95% of confidence interval (C.I). The overall seroprevalence was 65% (95% CI 62.5-72.9%), with variations according to age ranges. The prevalence of persistent serological positive corresponded to 37.7% (95% CI 34.2-41.5). The risk factors found by multivariate analyses were the lease of pastures (OR = 2.071 CI 1.485-3.690), the use of the same needle (OR = 2.249 CI 1.354-3.736), the molasses supplementation (OR = 2.742 CI 1.156-5.807), and the native Creole breed (OR = 1.895 CI 1.416-2.804). The results of this study confirmed the endemism and higher common exposure to BVDV, as well as the presence of serological persistent cattle in Valledupar, Colombia.

Effect of bovine viral diarrhea virus on subsequent infectivity of bovine gammaherpesvirus 4 in endometrial cells in primary culture: An in vitro model of viral co-infection.

Bovine viral diarrhea virus (BVDV) and bovine gammaherpesvirus 4 (BoHV-4) infect the uterus of cattle, being responsible for huge economic losses. Most of the pathogenesis of BoHV-4 in the bovine reproductive tract has been elucidated by conducting tests on primary cultures. Thus, it is important to have optimal in vitro conditions, avoiding the presence of other pathogens that can alter the results. BVDV is one of the most frequent viral contaminants of cell cultures. Considering that non-cytopathic (NCP) BVDV biotype can generate persistently infected (PI) cattle, which are the major source for virus transmission in susceptible herds, it is important to check products derived from cattle that are intended to be used in research laboratories. The aim of this work was to evaluate how the natural infection of bovine endometrial cells (BEC) with a NCP BVDV strain (BEC + BVDV) affects BoHV-4 replication. We have demonstrated a delay in BoHV-4 gene expression and a decrease in viral load in the extracellular environment in BEC + BDVD cells compared to BEC (BVDV-free) cells. These results confirm that replication of BoHV-4 in BEC primary cultures is affected by previous infection with BVDV. This finding highlights the importance of ruling out BVDV infection in bovine primary cell cultures to avoid biological interference or misinterpretation of results at the time of performing in vitro studies with BoHV-4.

Quantitative risk assessment of introduction of BVDV and BoHV-1 through indirect contacts based on implemented biosecurity measures in dairy farms of Spain.

A stochastic quantitative risk assessment model was developed to estimate the annual probability of introduction of bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) on 127 dairy farms through indirect contacts. Vehicles transporting calves, cattle to slaughterhouse, dead animals, and mixture of feed, as well as visits by veterinarians and hoof trimmers, farm workers and contacts with neighbors were considered in the model. Data from biosecurity questionnaires of each farm, scientific literature and expert opinion from field veterinarians, animal vehicle drivers, hoof trimmers and personnel from rendering transport companies were used to estimate values for input parameters. Results showed that the annual probability of introducing BVDV or BoHV-1 through indirect contacts was very heterogeneous. The overall distribution of median values for each farm ranged from 0.5 to 14.6% and from 1.0 to 24.9% for BVDV and BoHV-1, respectively. The model identified that providing protective clothing and boots to visits, not allowing the animal vehicle driver to come into contact with animals present on the farm and ensuring that calf vehicles arrived empty, were the measures with the highest impact on the probability of infection for most farms. This model could be a useful tool to show the impact of the measures to farmers and veterinarians, thus increasing their awareness on biosecurity. In addition, it could support decision making on which measures should be prioritized in dairy cattle herds to reduce the probability of introduction of diseases.

Upregulation of interferon-alpha gene in bovine embryos produced in vitro in response to experimental infection with noncytophatic bovine-viral-diarrhea virus.

In-vitro fertilization is a routine livestock-breeding technique widely used around the world. Several studies have reported the interaction of bovine viral-diarrhea virus (BVDV) with gametes and in-vitro-produced (IVP) bovine embryos. Since, gene expression in BVDV-infected IVP bovine embryos is scarcely addressed. The aim of this work was to evaluate the differential expression of genes involved in immune and inflammatory response. Groups of 20-25 embryos on Day 6 (morula stage) were exposed (infected) or not (control) to an NCP-BVDV strain in SOF medium. After 24 h, embryos that reached expanded blastocyst stage were washed. Total RNA of each embryo group was extracted to determine the transcription levels of 9 specific transcripts related with antiviral and inflammatory response by SYBR Green real time quantitative (RT-qPCR). Culture media and an aliquot of the last embryos wash on Day 7 were analyzed by titration and virus isolation, respectively. A conventional PCR confirmed BVDV presence in IVP embryos. A significantly higher expression of interferon-α was observed in blastocysts exposed to NCP-BVDV compared to the controls (p < 0.05). In this study, the upregulation of INFα and TLR7 genes involved in inflammatory and immune response in BVDV-infected IVP bovine embryos is a new finding in this field. This differential expression suggest that embryonic cells could function in a manner like immune cells by recognizing and responding early to interaction with viral pathogens. These results provide new insights into the action of BVDV on the complex molecular pathways controlling bovine early embryonic development.

Interferon lambda protects cattle against bovine viral diarrhea virus infection.

Interferon lambda (IFN-λ) plays an important role in inducing an antiviral state in mucosal surfaces and has been used as an effective biotherapeutic against several viral diseases. Here we performed a proof of concept study on the activity of a biologically active recombinant bovine IFN-λ (rIFN-λ) produced in eukaryotic cells against Bovine Viral Diarrhea Virus (BVDV) in cattle. We first confirmed the lack of toxicity of different concentrations of rIFN-λ in bovine peripheral blood cells and the safety of its subcutaneous application in calves in doses up to 12 IU/kg. The antiviral activity of the rIFN-λ against BVDV was assessed in calves that were inoculated with 6 IU/kg of rIFN-λ (n = 4) or mock-treated (n = 2) two days before and after challenge with a BVDV type-2 non-cytopathic strain. Mock-treated animals developed respiratory disease, shedded the virus from 4 to 7 days post-infection (dpi) and had viremia between 4 and 14 dpi. Conversely, calves treated with rIFN-λ did not develop clinical symptoms. The virus was not found in nasal secretions or sera. Only one animal had a positive viral RNA detection in serum at 7 dpi. All infected animals treated with rIFN-λ increased systemic type-I IFNs levels at 4 dpi. The antiviral treatment induced an earlier onset of the anti-BVDV neutralizing antibodies. Altogether, these results constitute the proof-of-principle of bovine IFN-λ as an antiviral biotherapeutic to protect cattle against the clinical disease caused by BVDV.

Prevalence and risk factors of bovine viral diarrhea in Colombian cattle.

BACKGROUND AND AIM: Bovine viral diarrhea virus (BVDV) is present in most cattle-raising countries around the world, and it has a negative economic impact in cattle herds. In Colombia, previous studies have estimated the prevalence of BVDV in specific locations. The aim of this study was to estimate the prevalence of BVDV in cattle herds located at several municipalities across the country and to identify the associated risk and protective factors. MATERIALS AND METHODS: A cross-sectional study was carried out to investigate the prevalence of BVDV in Colombian cattle populations at farm and animal-levels. A total of 387 herds and 8110 animals located in seven different departments were included in this study. RESULTS: An animal- and farm-level prevalence of 36% and 69%, respectively, were estimated. A high variation for the farm-level prevalence was found among the municipalities studied. Moreover, seropositive cattle to the infectious bovine rhinotracheitis virus (odds ratio (OR)=2.38, p=0.0479) and Neospora caninum (OR=3.15, p=0.0122) were more likely to be seropositive for BVDV, while the practice of burning dead animals at the farm was identified as a protective factor (OR=0.17, p=0.014). CONCLUSION: The prevalence of BVDV varied more at farm-level compared to animal-level. Two risk factors and one protective factor were identified. The results of the current study are essential to understand the epidemiology of BVDV in Colombia, and to formulate strategies in the region to mitigate the impact of this virus on the productive and reproductive indicators of cattle farms at the regional level.

Co-infection by Neopora caninum and bovine viral diarrhea virus in cattle from Rio Grande do Sul, Brazil, destined to exportation / Coinfecção por Neospora caninum e vírus da diarreia viral bovina em bovinos do Rio Grande do Sul, Brasil, destinados à exportação

Reproductive tests in cattle are of great economic importance, given the impact it can have on the production system and may be caused by agents. Neospora caninum and Bovine Viral Diarrhea virus (BVDV) are considered of great importance as reproductive and should be considered responsible for keeping animals persistently infected. The present study included 479 calf serum samples for export in the state of Rio Grande do Sul (RS). All samples were screened for BVDV by an ELISA antigen. BVDV antigen-positive ELISA samples were isolated from BVDV in cell culture. An indirect immunofluorescence (IFT) technique was used to detect anti-N. caninum antibodies. Of the 479 export-treated serum samples, 361 were positive for BVDV antigens by ELISA and/or viral isolation test (361/479-75.36%), and 109 IFT-positive samples for N. caninum (109/479-22.75%). Despite detection of antibodies anti-N. caninum did not differ statistically between naturally infected BVDV and non-BVDV infected animals suggesting that there is no interference of BVDV infection on infection or detection rate of animals with N. caninum, positive animals in viral isolation and high DO in BVDV-Ag ELISA. may present active disease and consequent immunosuppression, contributing to a potential reactivation of N. caninum.(AU)

Testes reprodutivos em bovinos são de grande importância econômica, dado o impacto que podem ter no sistema de produção e podem ser causados por agentes. O Neospora caninum e o vírus da Diarreia Viral Bovina (BVDV) são considerados de grande importância como reprodutivos e devem ser considerados responsáveis por manter os animais persistentemente infectados. O presente estudo incluiu 479 amostras de soro de bezerro para exportação no estado do Rio Grande do Sul (RS). Todas as amostras foram rastreadas para BVDV por um antígeno ELISA. As amostras de ELISA positivas para o antigénio BVDV foram isoladas a partir de BVDV em cultura de células. Uma técnica de imunofluorescência indireta (IFT) foi utilizada para detectar anticorpos anti-N caninum. Das 479 amostras de soro tratadas para exportação, 361 foram positivas para antígenos de BVDV por ELISA e/ou teste de isolamento viral (361/479-75,36%) e 109 amostras positivas para IFT para N. caninum (109/479-22,75%). Apesar da detecção de anticorpos anti-N. caninum não diferiu estatisticamente entre animais infectados naturalmente BVDV e não BVDV sugerindo que não há interferência da infecção pelo BVDV na infecção ou taxa de detecção de animais com N. caninum, animais positivos em isolamento viral e alta DO em BVDV-Ag ELISA, pode apresentar doença ativa e consequente imunossupressão, contribuindo para uma potencial reativação de N. caninum.(AU)