Comparative efficacy of modified-live and inactivated vaccines in boosting responses to bovine respiratory syncytial virus, bovine parainfluenza virus Type 3, and bovine coronavirus following neonatal mucosal priming of beef calves.

Objective: This study compared clinical and immunological responses to coinfection challenge of beef calves mucosally primed and differentially boosted with commercial combination vaccines containing antigens against bovine coronavirus (BCoV), bovine parainfluenza virus Type 3 (BPIV3), and bovine respiratory syncytial virus (BRSV). Animals: Nineteen commercial beef heifers. Procedure: At birth, calves were mucosally (IN) primed with modified-live virus (MLV) vaccines, differentially boosted by injection of either combination MLV (IN-MLV) or inactivated virus (IN-KV) vaccines at a mean age of 44 d, and then challenged by coinfection with BCoV, BPIV3, and BRSV at weaning. Results: Both groups were similarly protected from clinical disease and had anamnestic neutralizing antibody responses to all 3 viruses. The IN-KV group shed more BCoV, and less BPIV3 and BRSV, than the IN-MLV group. Conclusion: These data indicated similar clinical and immunological protection between IN-MLV and IN-KV; however, shed of virus varied. Clinical relevance: Whereas boosting with KV or MLV appeared to have similar efficacy, viral shed differences may affect disease control.

Efficacité comparative des vaccins vivants modifiés et inactivés pour stimuler les réponses au virus respiratoire syncytial bovin, au virus parainfluenza bovin de type 3 et au coronavirus bovin après amorçage via la muqueuse de veaux de boucherie nouveau-nés. Objectif: Cette étude a comparé les réponses cliniques et immunologiques à une co-infection de veaux de boucherie amorcés par voie muqueuse et différentiellement stimulés avec des vaccins combinés commerciaux contenant des antigènes contre le coronavirus bovin (BCoV), le virus parainfluenza bovin de type 3 (BPIV3) et le virus respiratoire syncytial bovin (BRSV). Animaux: Dix-neuf génisses de boucherie commerciales. Procédure: À la naissance, les veaux ont été vaccinés au niveau des muqueuses (IN) avec des vaccins à virus vivants modifiés (MLV), stimulés de manière différentielle par l'injection de vaccins combinés MLV (IN-MLV) ou de virus inactivés (IN-KV) à un âge moyen de 44 jours. puis provoqué par une co-infection avec BCoV, BPIV3 et BRSV au sevrage. Résultats: Les deux groupes étaient protégés de la même manière contre la maladie clinique et présentaient des réponses anamnestiques en anticorps neutralisants contre les 3 virus. Le groupe IN-KV a excrété plus de BCoV et moins de BPIV3 et de BRSV que le groupe IN-MLV. Conclusion: Ces données indiquent une protection clinique et immunologique similaire entre IN-MLV et IN-KV; cependant, l'excrétion du virus variait. Pertinence clinique: Alors que le rappel avec KV ou MLV semble avoir une efficacité similaire, les différences d'excrétion virale peuvent affecter la limitation de la maladie.(Traduit par Dr Serge Messier).

The MVA vector expressing the F protein of bovine respiratory syncytial virus is immunogenic in systemic and mucosal immunization routes.

Bovine respiratory syncytial virus (BRSV) affects both beef and dairy cattle, reaching morbidity and mortality rates of 60-80% and 20%, respectively. The aim of this study was to obtain a recombinant MVA expressing the BRSV F protein (MVA-F) as a vaccine against BRSV and to evaluate the immune response induced by MVA-F after systemic immunization in homologous and heterologous vaccination (MVA-F alone or combined with a subunit vaccine), and after intranasal immunization of mice. MVA-F administered by intraperitoneal route in a homologous scheme elicited levels of neutralizing antibodies similar to those obtained with inactivated BRSV as well as better levels of IFN-γ secretion. In addition, nasal administration of MVA-F elicited local and systemic immunity with a Th1 profile. This study suggests that MVA-F is a good candidate for further evaluations combining intranasal and intramuscular routes, in order to induce local and systemic immune responses, to improve the vaccine efficacy against BRSV infection.

Dynamics of neutralizing antibodies against Bovine respiratory syncytial virus in a dairy herd from Santa Fe Province, Argentina.

Bovine respiratory syncytial virus (BRSV) is one of the most relevant agents responsible for respiratory disease in cattle from both dairy and beef farms. BRSV is spread by horizontal contact causing a constant presence of seropositive animals that favors viral circulation throughout the year. Moreover, reinfections with BRSV are frequent between animals regardless of their age as BRSV does not confer long-lasting protective immunity. Several studies have demonstrated the circulation of BRSV in cattle from different regions of the world; however, little is known about the dynamics of BRSV infection in cows before and after they begin lactation. The aim of this work was to study the dynamics of BRSV neutralizing antibodies from birth up to 36 months of age in a closed dairy herd of Argentina specifically around the lactation period. Passive maternal antibodies against BRSV started to decrease monthly and became almost undetectable at 8 months of age. We detected two potential infection points at months 11 and 27 after birth, in which 30% and 45% of the animals showed seroconversion, respectively. Specifically, an increase in the proportion of seropositive cows after the start of lactation suggests that they became reinfected around the time they began lactating. We demonstrate the importance of understanding BRSV dynamics in a closed dairy herd to review the vaccination schedule of the animals to achieve protection against BRSV infection.

El virus respiratorio sincitial bovino (Bovine respiratory syncytial virus, [BRSV]) es uno de los principales agentes responsables de la enfermedad respiratoria en bovinos, tanto de tambos como de cría. El virus se transmite horizontalmente y causa la presencia constante de animales seropositivos, lo cual favorece la circulación viral a lo largo del año. A su vez, las reinfecciones por BRSV son frecuentes entre animales independientemente de su edad, dado que el virus no confiere inmunidad protectora a largo plazo. Numerosos estudios han demostrado la circulación de BRSV en bovinos de diferentes regiones del mundo, sin embargo, poco se conoce acerca de la dinámica de infección en vacas antes y después del inicio de la fase de lactancia. El objetivo de este trabajo fue estudiar la dinámica de anticuerpos neutralizantes anti- BRSV en vacas lecheras desde el nacimiento hasta los 36 meses de vida en un tambo cerrado de Argentina, específicamente, en el período de lactancia. Los anticuerpos pasivos específicos para BRSV comenzaron a declinar mensualmente hasta ser casi indetectables a los 6 meses. Detectamos dos potenciales puntos de infección a los meses 11 y 27 luego del nacimiento, momentos en los que el 30 y el 45% de los animales mostraron seroconversión, respectivamente. El incremento en la proporción de vacas seropositivas luego del comienzo de la lactancia sugiere que estas se reinfectaron en el inicio de dicha etapa. Demostramos la importancia de entender la dinámica de circulación del BRSV en un tambo cerrado, a fin de revisar el esquema de vacunación de los animales para que estén protegidos frente a la posible infección por este virus.

Genome-wide association study for response to vaccination in Angus calves1.

BACKGROUND: Bovine respiratory disease complex (BRDC) is one of the most important sources of loss within the beef cattle industry in the USA. Steps have been taken to reduce the incidence of BRDC through vaccination. Despite the effectiveness of vaccines, large proportions of cattle still experience morbidity and mortality. Identification of genomic regions that are associated with variation in response to vaccination would allow for the selection of individuals genetically predisposed to respond to vaccination based on specific markers, while heritability and accuracy estimates would help facilitate genomic selection. This in turn may lead to selection for beef cattle herds that may have lower incidence rate of BRDC after vaccination. This study utilizes an Angus herd of more than 2000 head of cattle to identify these regions of association. RESULTS: Genome wide association studies were performed for viral neutralization antibody level and response to vaccination traits against four different viruses associated with BRDC: bovine viral diarrhea virus 1 and 2 (BVDV1 and BVDV2), bovine respiratory syncytial virus (BRSV), and bovine herpesvirus (BHV1). A total of six 1-Mb windows were associated with greater than 1% of the genetic variance for the analyzed vaccination response traits. Heritabilities ranged from 0.08 to 0.21 and prediction accuracy ranged from 0.01 to 0.33 across 7 different vaccination traits. CONCLUSIONS: Although six 1-Mb windows were identified as associated with 1% or greater genetic variance for viral neutralization antibody level and response to vaccination traits, few genes around these windows could readily be considered candidates. This indicates the need for further functional genomic annotation, as these regions appear to be gene deserts. Traits ranged from lowly to moderately heritable, which indicated the potential for selection of individuals that are genetically pre-disposed to respond to vaccination. The relatively low amount of genetic variance accounted for by any 1-Mb window indicated that viral neutralization antibody level and response to vaccination traits are polygenic in nature. Selection for these traits is possible, but likely to be slow due to the low heritabilities and absence of markers with high genetic variation associated with them.

Cross-neutralization of four paramyxoviruses by a human monoclonal antibody.

Broadly neutralizing antibodies reactive against most and even all variants of the same viral species have been described for influenza and HIV-1 (ref. 1). However, whether a neutralizing antibody could have the breadth of range to target different viral species was unknown. Human respiratory syncytial virus (HRSV) and human metapneumovirus (HMPV) are common pathogens that cause severe disease in premature newborns, hospitalized children and immune-compromised patients, and play a role in asthma exacerbations. Although antisera generated against either HRSV or HMPV are not cross-neutralizing, we speculated that, because of the repeated exposure to these viruses, cross-neutralizing antibodies may be selected in some individuals. Here we describe a human monoclonal antibody (MPE8) that potently cross-neutralizes HRSV and HMPV as well as two animal paramyxoviruses: bovine RSV (BRSV) and pneumonia virus of mice (PVM). In its germline configuration, MPE8 is HRSV-specific and its breadth is achieved by somatic mutations in the light chain variable region. MPE8 did not result in the selection of viral escape mutants that evaded antibody targeting and showed potent prophylactic efficacy in animal models of HRSV and HMPV infection, as well as prophylactic and therapeutic efficacy in the more relevant model of lethal PVM infection. The core epitope of MPE8 was mapped on two highly conserved anti-parallel ß-strands on the pre-fusion viral F protein, which are rearranged in the post-fusion F protein conformation. Twenty-six out of the thirty HRSV-specific neutralizing antibodies isolated were also found to be specific for the pre-fusion F protein. Taken together, these results indicate that MPE8 might be used for the prophylaxis and therapy of severe HRSV and HMPV infections and identify the pre-fusion F protein as a candidate HRSV vaccine.

Comparative efficacy of modified-live and inactivated vaccines in boosting responses to bovine respiratory syncytial virus following neonatal mucosal priming of beef calves.

Bovine respiratory syncytial virus (BRSV) is the leading cause of viral pneumonia in calves, making young passively immune calves candidates for vaccination, and raising issues concerning boosting of neonatally primed responses. To address this, 18, 2-month-old Angus-cross passively immune beef heifer calves that had been primed at birth with a combination viral intranasal vaccine were administered either a parenteral combination vaccine containing modified-live (MLV) BRSV or a similar vaccine containing inactivated BRSV. At 6 months of age, these calves and 2 controls that received only the MLV at 2 months of age were challenged with BRSV via aerosol. Two calves, 1 control, and 1 MLV-boosted, developed severe respiratory disease and required euthanasia; the remaining calves developed no or mild respiratory disease and recovered. Calves that received the inactivated booster had significantly higher arterial oxygen concentrations on Day 7 after challenge and had anamnestic BRSV-specific IgG and neutralizing antibodies after challenge; the MLV-boosted calves did not. These data suggest that adjuvanted inactivated parenteral BRSV vaccines administered at 2 months of age may provide better boosting for neonatally mucosally primed calves.

Efficacité comparée des vaccins vivants modifiés et des vaccins inactivés pour améliorer la réponse au virus respiratoire syncytial bovin après la sensibilisation active néonatale des muqueuses chez les veaux de boucherie. Le virus respiratoire syncytial bovin (VRS) est la cause principale de pneumonie virale chez les veaux, ce qui rend des jeunes veaux à immunité passive des candidats pour la vaccination et soulève des enjeux liés à l'amélioration de la réponse des nouveau-nés sensibilisés. Dans le but d'aborder cette situation, 18 veaux de boucherie de race croisée Angus âgés de 2 mois ayant une immunité passive, qui avaient été sensibilisés activement à la naissance à l'aide d'une combinaison de vaccins intranasaux viraux, ont reçu soit un vaccin combiné parentéral contenant le VRS modifié vivant (VMV) ou un vaccin semblable contenant le VRS inactivé. À l'âge de 6 mois, ces veaux et deux témoins qui avaient reçu seulement le VNV à l'âge de 2 mois, ont été exposés au VRS par voie aérosol. Deux veaux, un témoin et un animal ayant reçu le rappel VMV, ont développé une maladie respiratoire grave et ont dû être euthanasiés; les autres animaux ont développé une maladie respiratoire légère et se sont rétablis ou n'ont manifesté aucun symptôme. Les veaux qui avaient reçu le rappel inactivé affichaient des concentrations d'oxygène significativement supérieures dans le sang artériel le jour 7 après le test et présentaient des anticorps neutralisants et anamnestiques spécifiques aux VRS après le test, contrairement aux veaux ayant reçu le rapport VNV. Ces données suggèrent que les vaccins VRS parentéraux inactivés avec adjuvants administrés à l'âge de 2 mois peuvent offrir une meilleure protection pour les veaux sensibilisés activement à la naissance sur les muqueuses.(Traduit par Isabelle Vallières).

Bovine respiratory syncytial virus-specific IgG-1 in nasal secretions of colostrum-fed neonatal calves.

In order to determine whether nasal secretions of young calves contain passively derived antibodies to bovine respiratory syncytial virus (BRSV) and if there are differences in presence and/or subclass of these antibodies between calves fed different colostrum replacement products, 17 Holstein calves were fed 150 g of IgG in either a sprayed-dried colostrum-based (CR; n = 8) or a plasma-based colostrum replacement product (PR; n = 9) within 6 h of birth. Venous blood and nasal secretions obtained before feeding and at 24 h of age were assayed for total IgG (serum) by radial immunodiffusion and for BRSV-specific total IgG, IgG-1, and IgG-2 by indirect enzyme-linked immunosorbent assay (ELISA). Calves that were fed a CR had higher concentrations of BRSV-specific IgG and IgG-1 in their serum and nasal secretions compared to calves fed product PR; calves fed the PR had higher levels of serum BRSV-specific IgG-2. The only subclass of antibodies detected in nasal secretions was IgG-1. Re-secretion of passive IgG with neutralizing activity, onto the nasal mucosa could contribute to BRSV-associated disease-sparing observed in the laboratory and in the field. Use of PR will result in lower nasal antibodies since IgG-2 is not re-secreted.

IgG-1 spécifique au virus respiratoire syncytial bovin dans les sécrétions nasales des veaux néonataux nourris au colostrum. Afin de déterminer si les sécrétions nasales des jeunes veaux contenaient des anticorps dérivés passivement envers le virus respiratoire syncytial bovin (VRS) et s'il y a des différences dans la présence et/ou la sous-catégorie de ces anticorps entre les veaux nourris avec différents produits de remplacement du colostrum, 17 veaux Holstein ont été nourris avec 150 g d'IgG soit sous forme de produit vaporisé-séché à base de colostrum (CR; n = 8) ou d'un produit de remplacement de colostrum à base de plasma (PR; n = 9) au cours des 6 premières heures après la naissance. Du sang veineux et des sécrétions nasales obtenus avant le nourrissage et à l'âge de 24 h ont été analysés pour obtenir la quantité d'IgG totale (sérum) par immunodiffusion radiale et le total des quantités d'IgG, d'IgG-1 et d'IgG-2 spécifiques au VRS par ELISA indirecte. Les veaux qui avaient été nourris d'un CR avaient des concentrations supérieures d'IgG et dIgG-1 spécifiques au VRS dans leur sérum et les sécrétions nasales comparativement aux veaux nourris de produits PR; les veaux nourris d'un PR avaient des niveaux supérieurs d'IgG-2 sérique spécifique au VRS. La seule sous-catégorie d'anticorps détectée dans les sécrétions nasales était l'IgG-1. La re-secrétion passive d'IgG avec de l'activité neutralisante sur les muqueuses nasales pourrait contribuer à l'immunité associée au VRS observée en laboratoire et sur le terrain. L'usage de PR produira des anticorps nasaux inférieurs vu que l'IgG-2 n'est pas re-secrété.(Traduit par Isabelle Vallières).

Modulation of the transcription factor NF-κB in antigen-presenting cells by bovine respiratory syncytial virus small hydrophobic protein.

Bovine respiratory syncytial virus (BRSV) is an important cause of respiratory disease in young cattle and is closely related to human RSV (HRSV), which causes severe respiratory disease in infants and the elderly. The RSV genome encodes a small hydrophobic (SH) protein with viroporin activity. Previous studies have shown that recombinant BRSV lacking the SH gene (rBRSVΔSH) is attenuated in the lungs, but not in the upper respiratory tract, of calves and mucosal vaccination with rBRSVΔSH induced long-lasting protective immunity. Attenuation of rBRSVΔSH may be due to the ability of this virus to induce an early innate response as rBRSVΔSH induces higher levels of pro-inflammatory cytokines than wild-type (wt) rBRSV. In this study, we investigated the effects of the BRSV SH protein on NF-κB p65 phosphorylation, a master step in the regulation of pro-inflammatory cytokines. Expression of SH resulted in the inhibition of NF-κB p65 phosphorylation in response to BRSV infection and extracellular lipopolysaccharide, and a reduction in the production of pro-inflammatory cytokines. In contrast, rBRSVΔSH does not inhibit NF-κB p65 phosphorylation in bovine antigen-presenting cells, including monocytes, macrophages and dendritic cells, resulting in increased expression of pro-inflammatory cytokines and increased activation of T cells compared to cells infected with wt BRSV. These findings highlight an important role for the BRSV SH protein in immune modulation.

Identification of systemic immune response markers through metabolomic profiling of plasma from calves given an intra-nasally delivered respiratory vaccine.

Vaccination procedures within the cattle industry are important disease control tools to minimize economic and welfare burdens associated with respiratory pathogens. However, new vaccine, antigen and carrier technologies are required to combat emerging viral strains and enhance the efficacy of respiratory vaccines, particularly at the point of pathogen entry. New technologies, specifically metabolomic profiling, could be applied to identify metabolite immune-correlates representative of immune protection following vaccination aiding in the design and screening of vaccine candidates. This study for the first time demonstrates the ability of untargeted UPLC-MS metabolomic profiling to identify metabolite immune correlates characteristic of immune responses following mucosal vaccination in calves. Male Holstein Friesian calves were vaccinated with Pfizer Rispoval® PI3 + RSV intranasal vaccine and metabolomic profiling of post-vaccination plasma revealed 12 metabolites whose peak intensities differed significantly from controls. Plasma levels of glycocholic acid, N-[(3α,5ß,12α)-3,12-Dihydroxy-7,24-dioxocholan-24-yl]glycine, uric acid and biliverdin were found to be significantly elevated in vaccinated animals following secondary vaccine administration, whereas hippuric acid significantly decreased. In contrast, significant upregulation of taurodeoxycholic acid and propionylcarnitine levels were confined to primary vaccine administration. Assessment of such metabolite markers may provide greater information on the immune pathways stimulated from vaccine formulations and benchmarking early metabolomic responses to highly immunogenic vaccine formulations could provide a means for rapidly assessing new vaccine formulations. Furthermore, the identification of metabolic systemic immune response markers which relate to specific cell signaling pathways of the immune system could allow for targeted vaccine design to stimulate key pathways which can be assessed at the metabolic level.

A bovine respiratory syncytial virus model with high clinical expression in calves with specific passive immunity.

BACKGROUND: Bovine respiratory syncytial virus (BRSV) is a major cause of respiratory disease in cattle worldwide. Calves are particularly affected, even with low to moderate levels of BRSV-specific maternally derived antibodies (MDA). Available BRSV vaccines have suboptimal efficacy in calves with MDA, and published infection models in this target group are lacking in clinical expression. Here, we refine and characterize such a model. RESULTS: In a first experiment, 2 groups of 3 calves with low levels of MDA were experimentally inoculated by inhalation of aerosolized BRSV, either: the Snook strain, passaged in gnotobiotic calves (BRSV-Snk), or isolate no. 9402022 Denmark, passaged in cell culture (BRSV-Dk). All calves developed clinical signs of respiratory disease and shed high titers of virus, but BRSV-Snk induced more severe disease, which was then reproduced in a second experiment in 5 calves with moderate levels of MDA. These 5 calves shed high titers of virus and developed severe clinical signs of disease and extensive macroscopic lung lesions (mean+/-SD, 48.3+/-12.0% of lung), with a pulmonary influx of inflammatory cells, characterized by interferon gamma secretion and a marked effect on lung function. CONCLUSIONS: We present a BRSV-infection model, with consistently high clinical expression in young calves with low to moderate levels of BRSV-specific MDA, that may prove useful in studies into disease pathogenesis, or evaluations of vaccines and antivirals. Additionally, refined tools to assess the outcome of BRSV infection are described, including passive measurement of lung function and a refined system to score clinical signs of disease. Using this cognate host calf model might also provide answers to elusive questions about human RSV (HRSV), a major cause of morbidity in children worldwide.

Recombinant bovine respiratory syncytial virus with deletion of the SH gene induces increased apoptosis and pro-inflammatory cytokines in vitro, and is attenuated and induces protective immunity in calves.

Bovine respiratory syncytial virus (BRSV) causes inflammation and obstruction of the small airways, leading to severe respiratory disease in young calves. The virus is closely related to human (H)RSV, a major cause of bronchiolitis and pneumonia in young children. The ability to manipulate the genome of RSV has provided opportunities for the development of stable, live attenuated RSV vaccines. The role of the SH protein in the pathogenesis of BRSV was evaluated in vitro and in vivo using a recombinant (r)BRSV in which the SH gene had been deleted. Infection of bovine epithelial cells and monocytes with rBRSVΔSH, in vitro, resulted in an increase in apoptosis, and higher levels of TNF-α and IL-1ß compared with cells infected with parental, wild-type (WT) rBRSV. Although replication of rBRSVΔSH and WT rBRSV, in vitro, were similar, the replication of rBRSVΔSH was moderately reduced in the lower, but not the upper, respiratory tract of experimentally infected calves. Despite the greater ability of rBRSVΔSH to induce pro-inflammatory cytokines, in vitro, the pulmonary inflammatory response in rBRSVΔSH-infected calves was significantly reduced compared with that in calves inoculated with WT rBRSV, 6 days previously. Virus lacking SH appeared to be as immunogenic and effective in inducing resistance to virulent virus challenge, 6 months later, as the parental rBRSV. These findings suggest that rBRSVΔSH may be an ideal live attenuated virus vaccine candidate, combining safety with a high level of immunogenicity.

Bovine model of respiratory syncytial virus infection.

Bovine respiratory syncytial virus (BRSV), which is an important cause of respiratory disease in young calves, is genetically and antigenically closely related to human (H)RSV. The epidemiology and pathogenesis of infection with these viruses are similar. The viruses are host-specific and infection produces a spectrum of disease ranging from subclinical to severe bronchiolitis and pneumonia, with the peak incidence of severe disease in individuals less than 6 months of age. BRSV infection in calves reproduces many of the clinical signs associated with HRSV in infants, including fever, rhinorrhoea, coughing, harsh breath sounds and rapid breathing. Although BRSV vaccines have been commercially available for decades, there is a need for greater efficacy. The development of effective BRSV and HRSV vaccines face similar challenges, such as the need to vaccinate at an early age in the presence of maternal antibodies, the failure of natural infection to prevent reinfection, and a history of vaccine-augmented disease. Neutralising monoclonal antibodies (mAbs) to the fusion (F) protein of HRSV, which can protect infants from severe HRSV disease, recognise the F protein of BRSV, and vice versa. Furthermore, bovine and human CD8(+) T-cells, which are known to be important in recovery from RSV infection, recognise similar proteins that are conserved between HRSV and BRSV. Therefore, not only can the bovine model of RSV be used to evaluate vaccine concepts, it can also be used as part of the preclinical assessment of certain HRSV candidate vaccines.

Inhibition of priming for bovine respiratory syncytial virus-specific protective immune responses following parenteral vaccination of passively immune calves.

The effect of maternal antibodies (MatAb) on immunological priming by neonatal parenteral vaccination for bovine respiratory syncytial virus (BRSV) was addressed for the first time in experimental infection in 34 Holstein calves. Both vaccinated and control calves developed moderate to severe respiratory disease characteristic of acute BRSV infection. There were no differences in clinical signs, BRSV shed, arterial oxygen concentrations, or mortality between vaccinated and control calves after BRSV challenge approximately 11 wk after vaccination. There were no anamnestic antibody or cytokine responses in the vaccinates after challenge. Lung lesions were extensive in both groups, and although there was a statistically significant (P = 0.05) difference between groups, this difference was considered not biologically significant. These data indicate that stimulation of protective immune responses was inhibited by maternal antibodies when a combination modified-live BRSV vaccine was administered parenterally to young passively immune calves. Alternate routes of administration or different vaccine formulations should be used to successfully immunize young calves with good passive antibody transfer.

Inhibition de l'amorçage pour les réponses immunitaires protectrices spécifiques pour le virus respiratoire syncytial bovin après la vaccination parentérale des veaux ayant une immunité passive. L'effet des anticorps maternels sur l'amorçage immunologique par une vaccination parentérale néonatale pour le virus respiratoire syncytial bovin (VRS) a été abordé pour la première fois dans une infection expérimentale chez 34 veaux Holstein. Les veaux vaccinés et témoins ont développé une maladie respiratoire de modérée à grave présentant les caractéristiques d'une infection aiguë au VRS. Il n'y avait pas de différences au niveau des signes cliniques, de l'excrétion du VRS, des concentrations d'oxygène artérielle ou de la mortalité entre les veaux vaccinés et témoins après un test de provocation de VRS, environ 11 semaines après le vaccin. Il n'y avait aucune réponse d'anticorps ou de cytokines anamnestiques chez les veaux vaccinés après le test de provocation. Les lésions aux poumons étaient importantes dans les deux groupes et, même s'il y avait une différence statistiquement significative (P = 0,05) entre ces groupes, cette différence n'était pas considérée significative sur le plan biologique. Ces données indiquent que la stimulation des réponses immunitaires protectrices a été inhibée par les anticorps maternels lors de l'administration parentérale d'une combinaison de vaccin à VRS vivant modifié aux jeunes veaux ayant une immunité passive. D'autres voies d'administration ou différentes formulations de vaccins devraient être utilisées pour immuniser avec succès les jeunes veaux ayant un bon transfert passif.(Traduit par Isabelle Vallières).

Intranasal booster vaccination of beef steers reduces clinical signs following experimental coinfection with BRSV and BHV-1 without reducing shedding of BRD-associated bacteria.

OBJECTIVE: To determine the efficacy of primary or booster intranasal vaccination of beef steers on clinical protection and pathogen detection following simultaneous challenge with bovine respiratory syncytial virus and bovine herpes virus 1. METHODS: 30 beef steers were randomly allocated to 3 different treatment groups starting at 2 months of age. Group A (n = 10) was administered a single dose of a parenteral modified-live vaccine and was moved to a separate pasture. Groups B (n = 10) and C (10) remained unvaccinated. At 6 months of age, all steers were weaned and transported. Subsequently, groups A and B received a single dose of an intranasal modified-live vaccine vaccine while group C remained unvaccinated. Group C was housed separately until challenge. Two days following vaccination, all steers were challenged with bovine respiratory syncytial virus and bovine herpes virus 1 and housed in a single pen. Clinical and antibody response outcomes and the presence of nasal pathogens were evaluated. RESULTS: The odds of clinical disease were lower in group A compared with group C on day 7 postchallenge; however, antibody responses and pathogen detection were not significantly different between groups before and following viral challenge. All calves remained negative for Histophilus somni and Mycoplasma bovis; however, significantly greater loads of Mannheimia haemolytica and Pasteurella multocida were detected on day 7 postchallenge compared with day -2 prechallenge. CLINICAL RELEVANCE: Intranasal booster vaccination of beef steers at 6 months of age reduced clinical disease early after viral challenge. Weaning, transport, and viral infection promoted increased detection rates of M haemolytica and P multocida regardless of vaccination status.

Differential chemokine and cytokine production by neonatal bovine γδ T-cell subsets in response to viral toll-like receptor agonists and in vivo respiratory syncytial virus infection.

γδ T cells respond to stimulation via toll-like receptors (TLR). Bovine γδ T cells express TLR3 and TLR7, receptors that are key for the recognition of viruses such as bovine respiratory syncytial virus (BRSV); however, responses of γδ T cells to stimulation via these receptors, and their role during viral infections, remains unclear. Here, we demonstrate that neonatal bovine γδ T cells exhibit robust chemokine and cytokine production in response to the TLR3 agonist, Poly(I:C), and the TLR7 agonist, Imiquimod. Importantly, we observe a similar phenotype in γδ T-cell subsets purified from calves infected with BRSV. Bovine γδ T cells are divided into subsets based upon their expression of WC1, and the response to TLR stimulation and viral infection differs between these subsets, with WC1.1(+) and WC1(neg) γδ T cells producing macrophage inflammatory protein-1α and granulocyte-macrophage colony-stimulating factor, and WC1.2(+) γδ T cells preferentially producing the regulatory cytokines interleukin-10 and transforming growth factor-ß. We further report that the active vitamin D metabolite 1,25-dihydroxyvitamin D3 does not alter γδ T-cell responses to TLR agonists or BRSV. To our knowledge, this is the first characterization of the γδ T-cell response during in vivo BRSV infection and the first suggestion that WC1.1(+) and WC1(neg) γδ T cells contribute to the recruitment of inflammatory populations during viral infection. Based on our results, we propose that circulating γδ T cells are poised to rapidly respond to viral infection and suggest an important role for γδ T cells in the innate immune response of the bovine neonate.

The effects of viral vaccination of dairy heifer calves on the incidence of respiratory disease, mortality, and growth.

Bovine respiratory disease (BRD) is one of the most common infectious causes of morbidity and mortality in young dairy cattle. The objective of this randomized clinical trial was to determine the effectiveness of 1 or 2 doses of a 5-way, modified-live viral vaccine, administered to heifer calves before weaning to aid in the prevention of BRD. The hypotheses were that vaccination would reduce the incidence of BRD and mortality, and that 2 doses would be more effective than 1. A total of 2,874 heifer calves from 19 commercial dairy farms in Minnesota and Ontario were enrolled at 1 to 7d of age and were followed until 3 mo of age. Calves were randomly assigned to receive a commercial, intramuscular, modified-live vaccine against bovine viral diarrhea virus types 1 and 2, bovine respiratory syncytial virus, bovine herpesvirus type 1, and parainfluenza virus type 3 at 15 to 21 d of age (2 wk only), 35 to 42 d (5 wk only), both 2 and 5 wk, or sterile saline at both times (unvaccinated controls). The incidence of failure of passive transfer was 11 or 32%, using cut-points of serum total protein of 5.2 and 5.7 g/dL, respectively. Overall, 22% of calves were treated at least once for BRD. The incidence risk of naturally occurring BRD was 7.7% before 2 wk of age, 8.0% between 2 and 5 wk, and 9.5% between 5 wk and 3 mo of age, and was not different between vaccination groups. Overall mortality throughout the 3-mo study period was 3.5%. Mortality was 1.6% before 2 wk of age, 0.5% between 2 and 5 wk, and 1.2% between 5 wk and 3 mo of age. The risk of mortality was not affected by vaccination. Mean average daily gain of 1.07 kg/d from 5 wk to 3 mo of age was not different between vaccine groups. In this population of commercial, home-raised calves, with an overall low incidence of failure of passive transfer, intramuscular vaccination with a multivalent, modified live viral vaccine at 2 or 5 wk of age or both was not associated with a decreased risk of BRD or mortality, or with growth until 3 mo of age. Reasons for these findings may include interference by maternal antibodies, unresponsiveness of the neonatal immune system, timing of immunity relative to pathogen exposure, disease caused by pathogens other than the viruses in the vaccine, or herd immunity. However, in populations with higher incidence of failure of passive transfer or risk of BRD, calves with low levels of specific antibodies may respond differently to vaccination.

Detection of antibodies and risk factors for infection with bovine respiratory syncytial virus and parainfluenza virus 3 in dual-purpose farms in Colima, Mexico.

A cross-sectional study was carried out, from November 2007 to March 2008, to estimate the prevalence of and to determine risk factors associated with bovine syncytial respiratory virus (BRSV) and parainfluenza 3 virus (PIV3) in dual-purpose herds in Colima, México. One hundred and seventy-six sera from 33 herds for PIV3 and 232 sera from 44 herds for BRSV were used. Sera were analyzed by indirect ELISA for the detection of antibodies against BRSV and PIV3 in cattle herds to determine the seroprevalence of respiratory diseases. The apparent and true prevalences for PIV3 were 60.8% and 54.4% and for BRSV 52.2% and 50.8%, respectively. The percentage of herds showing at least one positive animal was 78.7% for PIV3, and 93.2% for BRSV. Age (≤ 12, 13-48, and >48 months old) and respiratory signs (no, yes) showed significant association (P < 0.05) with PIV3 and age with BRSV. This study showed that animals were exposed to both viruses and that age was the main risk factor. The need to establish new vaccination plans to effectively protect cattle against those infections in the state of Colima, Mexico is suggested.

Prevalence of and risk factors for bovine respiratory syncytial virus (BRSV) infection in non-vaccinated dairy and dual-purpose cattle herds in Ecuador.

A cross-sectional study was carried out to determine the seroprevalence and risk factors associated with bovine respiratory syncytial virus (BRSV) infection in non-vaccinated dairy and dual-purpose cattle herds from Ecuador. A total of 2,367 serum samples from 346 herds were collected from June 2008 to February 2009. A questionnaire, which included variables related to cattle, health, management measures, and the environment, was filled out in each herd. Presence of antibodies against BRSV was analyzed using a commercial indirect ELISA test. A logistic regression model was used to determine risk factors associated with BRSV at herd level. The individual seroprevalence against BRSV in non-vaccinated herds in Ecuador was 80.48% [1,905/2,367; 95% confidence interval (CI) = 78.9-82.1]. The herd prevalence was 91.3% (316/346; 95% CI = 88.3-94.3), and the intra-herd prevalence ranged between 25% and 100% (mean, 90.47%). The logistic regression model showed that the existence of bordering cattle farms, the dual-purpose farms, and the altitude of the farm (more than 2,338 m above sea level) were risk factors associated with BRSV infection. This is the first study about BRSV prevalence in Ecuador. It shows the wide spread of the BRSV infection in the country. The risk factors found will help to design effective control strategies.

Comparison of the immune response following subcutaneous versus intranasal modified-live virus booster vaccination against bovine respiratory disease in pre-weaning beef calves that had received primary vaccination by the intranasal route.

This study was performed to elucidate whether the route of booster vaccination affects the immune response against respiratory vaccine viruses in pre-weaning beef calves that receive primary intranasal (IN) vaccination during the first month of life. The objective was to compare the serum neutralizing antibody (SNA) titers to BHV1, BRSV, and BPI3V, cytokine mRNA expression and mucosal BHV1- and BRSV-specific IgA in nasal secretions following administration of IN or subcutaneous (SC) modified-live virus (MLV) booster vaccines 60 days after primary IN vaccination in young beef calves. Twenty-one beef calves were administered 2 mL of an IN MLV vaccine containing BHV1, BRSV, and BPI3V (Inforce3®) between one and five weeks of age. Sixty days after primary vaccination, calves were randomly assigned to one of two groups: IN-MLV (n = 11): Calves received 2 mL of the same IN MLV vaccine used for primary vaccination and 2 mL of a SC MLV vaccine containing BVDV1 & 2 (Bovi- Shield GOLD® BVD). SC-MLV (n = 10): Calves were administered 2 mL of a MLV vaccine containing, BHV1, BRSV, BPI3V, and BVDV1 & 2 (Bovi-Shield GOLD® 5). Blood and nasal secretion samples were collected on days -61 (primary vaccination), -28, -14, 0 (booster vaccination), 14, 21, 28, 42 and 60 for determination of SNA titers, cytokine gene expression analysis and nasal virus-specific IgA concentrations. Statistical analysis was performed using a repeated measures analysis through PROC GLIMMIX of SAS®. Booster vaccination by neither IN nor SC routes induced a significant increase in SNA titers against BHV1, BRSV, and BPI3V. Subcutaneous booster vaccination induced significantly greater BRSV-specific SNA titers (on day 42) and IgA concentration in nasal secretions (on days 21 and 42) compared to calves receiving IN booster vaccination. Both IN and SC booster vaccination were able to stimulate the production of BHV1-specific IgA in nasal secretions. In summary, booster vaccination of young beef calves using either SC or IN route two months after IN MLV primary vaccination resulted in comparable SNA titers, cytokine gene expression profile and virus-specific IgA concentration in nasal secretions. Only a few differences in the systemic and mucosal immune response against BHV1 and BRSV were observed. Subcutaneous booster vaccination induced significantly greater BRSV-specific SNA and secretory IgA titers compared to IN booster vaccination.

Impact of meloxicam on respiratory virus titers and health outcomes when administered concurrently with a modified live respiratory vaccine in abruptly weaned beef steers.

Abruptly weaned crossbred steer calves (N = 271) were used in a randomized, blinded 2-arm clinical trial to assess the impact of a long-acting non-steroidal anti-inflammatory drug on bovine herpesvirus type 1, bovine respiratory syncytial virus, parainfluenza virus type 3, and coronavirus titers and health outcomes when administered concurrently with a modified live respiratory vaccine upon arrival at a feedlot. Treatment groups included a control (saline; n = 135) and an experimental group (injectable meloxicam; n = 136). Viral antibody titers and body weight were measured on arrival, day 7, and day 21, along with a final weight on day 45. Body weight and antibody titers for all viruses increased over time (P < 0.001); however, there were no differences by treatment group or a significant group × time interaction when evaluated using repeated measures analysis of variance. Interestingly, the use of meloxicam was associated with increased treatment risk (P < 0.05). In conclusion, the administration of meloxicam may adversely affect health; however, a decreased vaccine response is likely not a contributing factor.

Des bouvillons croisés sevrés rapidement (N = 271) ont été utilisés dans un essai clinique randomisé en aveugle à deux bras pour évaluer l'impact d'un anti-inflammatoire non stéroïdien à action prolongée sur les titres du virus de la rhinotrachéite infectieuse bovine, du virus respiratoire syncytial bovin, du virus parainfluenza 3 et du coronavirus, et les résultats pour la santé lorsqu'administré en même temps qu'un vaccin vivant modifié respiratoire à l'arrivée dans un parc d'engraissement. Les groupes de traitement comprenaient un témoin (solution saline; n = 135) et un groupe expérimental (méloxicam injectable; n = 136). Les titres d'anticorps viraux et le poids corporel ont été mesurés à l'arrivée, au jour 7 et au jour 21, ainsi qu'un poids final au jour 45. Le poids corporel et les titres d'anticorps pour tous les virus ont augmenté avec le temps (P < 0,001); cependant, il n'y avait aucune différence selon le groupe de traitement ou une interaction groupe × temps significative lors de l'évaluation à l'aide de mesures répétées d'analyse de la variance. Fait intéressant, l'utilisation du méloxicam était associée à un risque de traitement accru (P < 0,05). En conclusion, l'administration de méloxicam peut nuire à la santé; cependant, une réponse vaccinale réduite n'est probablement pas un facteur contributif.(Traduit par Docteur Serge Messier).