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Many questions are raised, and challenges faced in the new era of (intranasal) bovine respiratory disease complex vaccination. An increase in vaccination rate is expected, due to its positive impact on cattle health, reduction of antimicrobial use and economic factors. However, engagement of farmers and veterinarians with regard to vaccination is often affected by limitations, resulting in the development of barriers to vaccination, but also opportunities to overcome these. The objective of the report is to provide practical recommendations and a consensus on best practises for BRDC vaccination, addressing barriers faced by veterinarians and farmers. The report combines an evidence review with expert opinions and includes discussions on different vaccination approaches, such as intranasal and systemic protocols. As result of the discussions, several barriers to BRDC vaccination were identified, including a lack of knowledge or visibility of the disease's impact, the preference for blanket antibiotic use over vaccination, resistance to change, the need for visible success, uncertainty about the best time to vaccinate, and concerns about adverse reactions and vaccine efficacy in the presence of maternal antibodies. While these barriers seem substantial, they provide opportunities for the veterinary sector. Indeed, veterinarians are encouraged to use the argumentation presented, along with local case studies and diagnostic testing to highlight the impact of disease, while conducting calf health audits, ensuring expectations are managed to achieve visible success. Overall, this consensus paper aims to provide practical recommendations and support for veterinarians and farmers to overcome barriers and increase BRDC vaccination rates in cattle.
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Abstract. Background: Infectious bovine respiratory disease complex (BRDC) is one of the world's major livestock problems. Aims: The study aimed to determine the diagnostic importance of pentraxin-3, endothelin-1, clinical biochemistry, and hematological parameters in infectious BRDC. Methods: Animals in this study were Simmental breed, 1-7 years old, untreated, and healthy and BRDC cattle (40 cattle with BRDC in the disease group, and 10 healthy cattle in the control group). Clinical findings such as general posture, respiratory rate per minute, rectal temperature, heart rate per minute, and mental posture of the diseased cattle were recorded. Blood samples were taken from the jugular vein only once from all cattle. Complete blood count from blood samples was measured in an automatic complete blood count device, biochemical parameters in an autoanalyzer, and pentraxin-3 and endothelin-1 were measured by ELISA method. Results: Rectal temperature, respiratory and pulse rates per minute, total leukocyte count, gamma-glutamyl transferase, urea, total bilirubin, lactate dehydrogenase, creatine kinase, pentraxin-3 and endothelin-1 concentrations were found to be statistically higher in BRDC group than those in the control group (P<0.001). Conclusion: Pentraxin-3 and endothelin-1 levels were statistically significantly higher in the BRDC group compared to the control group. As a result, pentraxin-3 and endothelin-1 were found to be diagnostically important in cattle diagnosed with BRDC.
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Qualitative and quantitative PCR-based tests are widely used in both diagnostics and research to assess the prevalence of disease-causing pathogens in veterinary medicine. The efficacy of these tests, usually measured in terms of sensitivity and specificity, is critical in confirming or excluding a clinical diagnosis. We undertook a meta-analysis to assess the inherent value of published PCR diagnostic approaches used to confirm and quantify bacteria and viruses associated with bovine respiratory disease (BRD) in cattle. This review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A thorough search of nine electronic databases (Web of Science, EBSCOhost, Cambridge journals online, ProQuest, PubMed, Sage journals online, ScienceDirect, Wiley online library and MEDLINE) was undertaken to find studies that had reported on the use of PCR and/or qPCR for the detection and/or quantification of BRD associated organisms. All studies meeting the inclusion criteria for reporting quantitative PCR for identification of BRD associated microorganisms were included in the analysis. Studies were then assessed on the applications of the Minimum Information for Publication of Quantitative Real-Time PCR Experiment (MIQE) and PCR primer/probe sequences were extracted and tested for in silico specificity using a high level of stringency. Fourteen full-text articles were included in this study. Of these, 79% of the analysed articles did not report the application of the MIQE guidelines in their study. High stringency in silico testing of 144 previously published PCR primer/probe sequences found many to have questionable specificity. This review identified a high occurrence of primer/probe sequences with a variable in silico specificity such that this may have implications for the accuracy of reporting. Although this analysis was only applied to one specific disease state, identification of animals suspected to be suffering from bovine respiratory disease, there appears to be more broadly a need for veterinary diagnostic studies to adopt international best practice for reporting of quantitative PCR diagnostic data to be both accurate and comparable between studies and methodologies.
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Bovine respiratory disease complex (BRDC) occurs widely in cattle farms. The main viral pathogens include bovine viral diarrhea virus (BVDV), Bovine herpesvirus 1 (BoHV-1), bovine parainfluenza virus type 3 (BPIV3), and bovine respiratory syncytial virus (BRSV), and the newly emerged influenza D virus (IDV). In this study, we have developed a one-step multiplex real-time Polymerase Chain Reaction (PCR) capable of simultaneously detecting these five viral pathogens causing BRDC. The established assay could specifically detect targeted viruses without cross-reaction with others. The detection limit was ~10 copies/reaction for single real-time PCR and 100 copies/ reaction for multiplex real-time PCR assay. A total of 213 nasal samples from cattle with signs of respiratory tract disease were then collected for performance evaluation of the established platform, proving that the method has good specificity and sensitivity. The surveillance data suggested that BVDV and BoHV-1 infections are the dominant cause of BRDC in the herd, whereas the detection rate of IDV, BIPV3, and BRSV is relatively lower. In summary, the established assay provides technical support for rapid clinical detection of BRDC associated viral pathogens to guide the formulation of BRDC prevention and control measures.
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Bovine respiratory disease complex (BRDC) is a multifactorial disease of cattle which presents as bacterial and viral pneumonia. The causative agents of BRDC work in synergy to suppress the host immune response and increase the colonisation of the lower respiratory tracts by pathogenic bacteria. Environmental stress and/or viral infection predispose cattle to secondary bacterial infections via suppression of key innate and adaptive immune mechanisms. This allows bacteria to descend the respiratory tract unchallenged. BRDC is the costliest disease among feedlot cattle, and whilst vaccines exist for individual pathogens, there is still a lack of evidence for the efficacy of these vaccines and uncertainty surrounding the optimum timing of delivery. This review outlines the immunosuppressive actions of the individual pathogens involved in BRDC and highlights the key issues in the development of vaccinations against them.
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As a multifactor disease, the bovine respiratory disease complex (BRDC) causes high morbidity and mortality that is devastating to the cattle industry. To assess viral infections in beef cattle suffering from respiratory diseases in Inner Mongolia, 302 nasal swabs and serum samples were randomly collected from cattle with mild respiratory symptoms between March 2018 and May 2019. Our results showed that the rate of RT-PCR results positive for nucleic acids of viral pathogens in 6 cities was between 54 and 80%.The rates of bovine viral diarrhea virus (BVDV), bovine herpesvirus 1 (BHV-1), bovine parainfluenza virus type 3(BPIV3), and bovine respiratory syncytial virus(BRSV)infections were 44.70% (135/302), 24.83% (75/302), 5.63% (17/302), and 6.95% (21/302),respectively. There are also 8.94% (27/302) of samples were positive for BVDV and BHV-1, and 3.97% (12/302) of samples were positive for BPIV3 and BRSV. In addition, the RT-PCR products were sequenced, and phylogenetic analysis based on these sequences was performed. The results indicated that: a) all of the BVDV isolates were BVDV-1 and were classified as BVDV-1a (66.67%) and BVDV-1b (33.33%); b) all of the BHV-1 isolates were classified as subtype 1.1; 44.44% of the isolates were closely related to modified live viral vaccine strains, and 55.56% of the isolates were closer to epidemic strains; c) all of the BPIV3 isolates belonged to BPIV3c; d) all of the BRSV isolates were classified into subgroup III. It is suggested that an important cause of respiratory diseases for beef cattle is viral infection, and phylogenetic analysis can help us choose the proper strain to develop a vaccine.
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Complejo Respiratorio Bovino , Enfermedades de los Bovinos , Virus de la Diarrea Viral Bovina Tipo 1 , Virus Sincitial Respiratorio Bovino , Animales , Complejo Respiratorio Bovino/epidemiología , Bovinos , Enfermedades de los Bovinos/epidemiología , China/epidemiología , Filogenia , Virus Sincitial Respiratorio Bovino/genéticaRESUMEN
Bovine respiratory disease (BRD) is the costliest disease affecting the cattle industry globally. Orthomyxoviruses, influenza C virus (ICV) and influenza D virus (IDV) have recently been implicated to play a role in BRD. However, there are contradicting reports about the association of IDV and ICV to BRD. Using the largest cohort study (cattle, n = 599) to date we investigated the association of influenza viruses in cattle with BRD. Cattle were scored for respiratory symptoms and pooled nasal and pharyngeal swabs were tested for bovine viral diarrhea virus, bovine herpesvirus 1, bovine respiratory syncytial virus, bovine coronavirus, ICV and IDV by real-time PCR. Cattle that have higher viral loads of IDV and ICV also have greater numbers of co-infecting viruses than controls. More strikingly, 2 logs higher IDV viral RNA in BRD-symptomatic cattle that are co-infected animals than those infected with IDV alone. Our results strongly suggest that ICV and IDV may be significant contributors to BRD.
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Complejo Respiratorio Bovino/virología , Gammainfluenzavirus/patogenicidad , Infecciones por Orthomyxoviridae/veterinaria , Thogotovirus/patogenicidad , Carga Viral/veterinaria , Animales , Complejo Respiratorio Bovino/epidemiología , Bovinos , Coinfección/epidemiología , Coinfección/veterinaria , Coinfección/virología , Femenino , Gammainfluenzavirus/aislamiento & purificación , Ganado , Masculino , Oportunidad Relativa , Infecciones por Orthomyxoviridae/epidemiología , Infecciones por Orthomyxoviridae/virología , Prevalencia , ARN Viral/análisis , Thogotovirus/aislamiento & purificaciónRESUMEN
Coinfection by bovine respiratory syncytial virus (BRSV) and Pasteurella multocida (PM) frequently has been observed in cattle that develop severe pneumonia. We recently reported that BRSV infection significantly increased PM adherence to bovine lower respiratory tract epithelial cells. However, the molecular mechanisms of enhanced PM adherence are not completely understood. To investigate whether BRSV infection regulates any cellular adherence receptors on bovine bronchus- and lung-epithelial cells, we performed proteomic and functional analyses. The proteomic analysis showed that BRSV infection increased the accumulation of the platelet-activating factor receptor (PAFR) in both cell types. Molecular experiments, including specific blockade, knockdown, and overexpression of PAFR, indicated that PM adherence to these cell types depended on PAFR expression. These findings highlight the role, in cattle with severe pneumonia, of the synergistic effect of coinfection by BRSV and PM in the lower respiratory tract.
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Neonatal calves are highly susceptible to a number of diseases including those that infect via the mucosal surfaces of the respiratory and gastrointestinal tracts. In order to determine appropriate vaccine design and delivery systems, or to identify suitable immunostimulatory methods to combat these infections, a detailed understanding of the immune cell populations present at clinically relevant sites is key. Few studies have assessed the immune cell composition of the neonatal calf lung and comparisons with circulating immune cells in the blood are lacking. We describe immune cell populations present in the peripheral blood, bronchoalveolar lavage (BAL) fluid and lung tissue of young disease-free calves. Flow cytometric analysis revealed significant differences in cell subset distribution between the peripheral blood and respiratory tract, and between compartments within the respiratory tract. Notably, whereas WC1+ γδ TCR + T lymphocytes dominate the peripheral blood, both the BAL fluid and lung tissue contained a high proportion of myeloid cells which expressed CD14 and CD172a (SIRPα). Very low numbers of tissue myeloid cells expressed MHC Class II in comparison to circulating myeloid cells in the blood. Respiratory tract tissues had low frequencies of CD4+ and CD8 + T lymphocytes, which were significantly lower than in the blood. Differences in the proportion of NKp46+ natural killer cells were also observed between tissue compartments. In order to target vaccines or immunostimulatory therapeutics appropriately, these differences in immune cell populations in tissue compartments should be taken into consideration.
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Líquido del Lavado Bronquioalveolar/inmunología , Recuento de Linfocitos/veterinaria , Sistema Respiratorio/inmunología , Subgrupos de Linfocitos T/inmunología , Animales , Animales Recién Nacidos/inmunología , Líquido del Lavado Bronquioalveolar/citología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Bovinos , Citometría de Flujo , Células Asesinas Naturales/inmunología , Pulmón/citología , Pulmón/inmunología , Masculino , Sistema Respiratorio/citologíaRESUMEN
The synergistic infection of bovine respiratory syncytial virus (BRSV) and Pasteurella multocida (PM) may predispose cattle to develop severe pneumonia. Previously, we reported that BRSV infection significantly decreased PM adherence to the upper respiratory epithelial cells. It may allow bacteria to invade into the lower respiratory tract and lead to severe pneumonia. To investigate whether BRSV infection regulates the cell surface adherence receptor on bovine trachea epithelial cells (bTECs), we performed proteomic and functional analyses. BRSV infection decreased the expression of intercellular adhesion molecule-1 (ICAM1) on bTECs. Inhibition and knockdown experiments using anti-ICAM1 antibody and siRNAs targeting ICAM1 indicated that PM adherence to bTECs was dependent on ICAM1 expression. These data suggest that under normal conditions bTECs may capture PM in the upper respiratory tract, while BRSV infection reverses this mechanism. The proposed gateway function of bTECs is disrupted by BRSV infection that may facilitate bacterial invasion into the lower respiratory tract and lead to secondary or more severe respiratory infection.
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Adhesión Bacteriana , Células Epiteliales/microbiología , Células Epiteliales/virología , Molécula 1 de Adhesión Intercelular/genética , Pasteurella multocida/fisiología , Virus Sincitial Respiratorio Bovino/fisiología , Animales , Bronquios/citología , Bronquios/microbiología , Bronquios/virología , Bovinos , Células Cultivadas , Regulación hacia Abajo , Pulmón/citología , Pulmón/microbiología , Pulmón/virología , Interacciones Microbianas , Proteómica , Tráquea/citología , Tráquea/microbiología , Tráquea/virologíaRESUMEN
Bovine respiratory disease complex is a major disease affecting the global cattle industry. Multiple infections by viruses and bacteria increase disease severity. Previously, we reported that bovine respiratory syncytial virus (BRSV) infection increases adherence of Pasteurella multocida to human respiratory and bovine kidney epithelial cells. To examine the interaction between the virus and bacteria in bovine respiratory cells, we generated respiratory epithelial cell lines from bovine trachea (bTEC), bronchus (bBEC), and lung (bLEC). Although all established cell lines were infected by BRSV and P. multocida susceptibility differed according to site of origin. The cells derived from the lower respiratory tract (bBEC and bLEC) were significantly more susceptible to BRSV than those derived from the upper respiratory tract (bTEC). Pre-infection of bBEC and bLEC with BRSV increased adherence of P. multocida; this was not the case for bTEC. These results indicate that BRSV may reproduce better in the lower respiratory tract and encourage adherence of bacteria. Thus, we identify one possible mechanism underlying severe pneumonia.
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Coinfección/veterinaria , Células Epiteliales , Interacciones Microbianas , Infecciones por Pasteurella/veterinaria , Infecciones por Virus Sincitial Respiratorio/veterinaria , Animales , Complejo Respiratorio Bovino/microbiología , Complejo Respiratorio Bovino/virología , Bronquios/citología , Bronquios/microbiología , Bronquios/virología , Bovinos , Línea Celular , Células Cultivadas , Coinfección/microbiología , Coinfección/virología , Citocinas/metabolismo , Células Epiteliales/microbiología , Células Epiteliales/virología , Pulmón/citología , Pulmón/microbiología , Pulmón/virología , Infecciones por Pasteurella/virología , Pasteurella multocida/genética , Pasteurella multocida/aislamiento & purificación , Infecciones por Virus Sincitial Respiratorio/microbiología , Virus Sincitial Respiratorio Bovino/genética , Virus Sincitial Respiratorio Bovino/aislamiento & purificación , Tráquea/citología , Tráquea/microbiología , Tráquea/virologíaRESUMEN
We identified influenza C virus (ICV) in samples from US cattle with bovine respiratory disease through real-time PCR testing and sequencing. Bovine ICV isolates had high nucleotide identities (≈98%) with each other and were closely related to human ICV strains (≈95%). Further research is needed to determine bovine ICV's zoonotic potential.
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Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/virología , Gammainfluenzavirus/clasificación , Gammainfluenzavirus/genética , Infecciones por Orthomyxoviridae/veterinaria , Infecciones del Sistema Respiratorio/veterinaria , Animales , Bovinos , Enfermedades de los Bovinos/historia , Historia del Siglo XXI , Filogenia , Estados Unidos/epidemiología , Proteínas de la Matriz Viral/genéticaRESUMEN
Primary infection with bovine respiratory syncytial virus (BRSV) predisposes cattle to secondary infection with bacteria that cause bovine respiratory disease complex (BRDC). However, the interaction between BRSV and bacteria is unclear. This in vitro study examined the adherence of Pasteurella multocida (PM) to BRSV-infected cells was assessed in colony forming unit assays, by flow cytometry analysis, and by indirect immunofluorescence analysis (IFA) of epithelial cells (A549, HEp-2, and MDBK). An in vitro model based on infection of BRSV-infected epithelial cells revealed that PM adherence to BRSV-infected cells was 2- to 8-fold higher than uninfected cells. This was confirmed by flow cytometry analysis and IFA. Epithelial cell expression of mRNA encoding cytokines and chemokines increased after exposure to PM, but increased further after co-infection with BRSV and PM. BRSV-mediated adherence of PM to epithelial cells may underlie the serious symptoms of BRDC.
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Complejo Respiratorio Bovino/virología , Células Epiteliales/microbiología , Interacciones Microbianas , Pasteurella multocida/fisiología , Infecciones por Virus Sincitial Respiratorio/veterinaria , Células A549 , Animales , Sitios de Ligazón Microbiológica , Bovinos , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/virología , Células Epiteliales/virología , Citometría de Flujo , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Infecciones por Virus Sincitial Respiratorio/complicaciones , Infecciones por Virus Sincitial Respiratorio/virología , Virus Sincitial Respiratorio Bovino/aislamiento & purificación , Virus Sincitial Respiratorio Bovino/fisiología , Sistema Respiratorio/citología , Sistema Respiratorio/microbiologíaRESUMEN
Bovine viral diarrhea virus (BVDV) infections, whether as acute, persistent or contributing to co-infections, result in significant losses for cattle producers. Although, BVDV can be identified readily by real-time PCR and ELISA, detection and quantification of viral infection at the single cell level is extremely difficult. Detection at the single lymphoid cell level is important due to the immunomodulation that accompanies BVDV infection. A novel PrimeFlow RNA assay using in-situ detection of BVDV was evaluated. The model used to develop this technique included three BL-3 cell lines with different infection statuses, one not infected with BVDV, one infected with BVDV and one dual infected with BVDV and bovine leukosis virus. Using RNA probes specific for the BVDV-2a Npro-Erns coding region, BVDV RNA was detected from both contaminated BL-3 cell lines by flow cytometry and fluorescent microscopy. This is the first report on in-situ detection of BVDV at the single-cell level.
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Virus de la Diarrea Viral Bovina/aislamiento & purificación , Citometría de Flujo/métodos , Linfocitos/virología , Microscopía Fluorescente/métodos , ARN Viral/análisis , Análisis de la Célula Individual/métodos , Animales , Bovinos , Virus de la Diarrea Viral Bovina/genética , Virología/métodosRESUMEN
Bovine respiratory disease complex (BRDc) is a multi-factorial disease, involving both viral and bacterial pathogens, that negatively impacts the cattle feedlot industry. A nitric oxide releasing solution (NORS) has been developed and shown to have potential in the prevention of BRDc. This study investigated the underlying immunological mechanisms through which the nitroslyating agent NORS provides protection against the development of BRDc in susceptible cattle. An in vitro BRDc experimental model was designed using bovine peripheral blood mononuclear cells (PBMCs) which were infected with bovine herpesvirus 1 (BHV-1) and subsequently cultured with lipopolysaccharides (LPS) extracted from Mannheimia haemolytica bacteria. The cells were treated with NORS following viral infection to reflect the timing of administering the NORS treatment in feedlots during initial processing. An expression and protein analysis of key genes involved in the innate immune response was carried out. The BRDc model produced significant increases in gene expression (p<0.01) and protein release (p<0.05) of the proinflammatory cytokines IL-1ß and TNF. Treatment with NORS reduced the protein levels of IL-1ß (0.39-fold↓) (p<0.05) and TNF (0.48-fold↓) (p<0.01) in the BRDc experimental group when compared against the non-treatment BRDc controls. TLR4 expression, having been significantly reduced under the BRDc experimental conditions (0.33-fold↓) (p<0.05), increased significantly (0.76-fold↑) (p<0.05) following NORS treatment. This study provides evidence suggesting that NO may protect against the development of BRDc by limiting deleterious inflammation while simultaneously increasing TLR4 expression and enhancing the ability of the host to detect and respond to bacterial pathogens.
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Complejo Respiratorio Bovino/inmunología , Herpesvirus Bovino 1/inmunología , Leucocitos Mononucleares/fisiología , Lipopolisacáridos/toxicidad , Mannheimia haemolytica/metabolismo , Óxido Nítrico/metabolismo , Animales , Bovinos , Células Cultivadas , Citocinas/genética , Regulación de la Expresión Génica/inmunología , Lipopolisacáridos/inmunología , Mannheimia haemolytica/inmunologíaRESUMEN
Mannheimia haemolytica (MH) is a major cause of respiratory disease in bovine complex (BRDC) that causes great economic losses. To this day, there are no reports of birds affected by this bacterium. The present report describes an atypical outbreak of mannheimiosis in captive rheas (Rhea americana).(AU)
Mannheimia haemolytica (MH) é uma das principais causas da doença do complexo respiratório dos bovinos (DCRB) que causa grandes perdas econômicas. Até os dias atuais, não há relato de aves acometidas por tal bactéria. Relata-se um surto atípico de mannheimiose em emas em cativeiro (Rhea americana).(AU)
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Animales , Enfermedades de las Aves/epidemiología , Mannheimia haemolytica , Reiformes/microbiología , Enfermedades Respiratorias/veterinariaRESUMEN
Symptoms of bovine viral diarrhea virus (BVDV) infection range from subclinical to severe, depending on strain virulence. Several in vitro studies showed BVDV infection impaired leukocyte function. Fewer studies have examined the effects of in vivo BVDV infection on monocyte/macrophage function, especially with strains of differing virulence. We characterized cytokine production by bovine myeloid cells isolated early or late in high (HV) or low virulence (LV) BVDV2 infection. Given BVDV infection may enhance susceptibility to secondary bacterial infection, LPS responses were examined as well. Monocytes from HV and LV infected calves produced higher levels of cytokines compared to cells from controls. In contrast, monocyte-derived macrophage cytokine levels were generally reduced. Modulated cytokine expression in HV BVDV2 macrophages was associated with decreased MyD88 expression, likely due to its interaction with viral NS5A. These data and those of others, suggest that certain Flaviviridae may have evolved strategies for subverting receptor signaling pathways involving MyD88.