Bovine Respiratory Syncytial Virus Research Articles

Cross-sectional study of primary antimicrobial treatment and vaccination coverage in outbreaks of bovine respiratory disease on dairy and beef farms in northern Belgium.

To what extent veterinarians active in the dairy or beef sector follow the antimicrobial therapy guidelines made available in different European countries for bovine respiratory disease (BRD) outbreaks, and whether differences in therapeutic or preventive preferences for BRD management exist, is currently unknown. Therefore, the objectives of this cross-sectional study were to compare vaccination coverage and primary antimicrobial and anti-inflammatory treatment on dairy, beef and mixed-breed farms in northern Belgium, and determine their compliance with the recommendations made by the Belgian formulary. Information on antimicrobial and anti-inflammatory drug use and vaccination coverage from 190 BRD outbreaks in 180 herds, submitted by 101 veterinarians, was analysed. Multivariable linear probability models, adjusted for clustering at the veterinarian level, were used to determine differences between dairy and beef farms. Antimicrobials and non-steroidal anti-inflammatory drugs (NSAIDs) were used in 93.5% and 81.7% of the BRD outbreaks, respectively. First-line antimicrobials were used as primary treatment in only 42.3%, 50.9% and 38.6% of dairy, beef and mixed-breed farms, respectively. Significant differences (p<0.05) were observed between dairy and beef farms in terms of use of long-acting macrolides (-17.2 percentage points [pp]; 95% confidence interval [CI]: -31.9, -2.5), steroidal anti-inflammatory drugs (15.2 pp; CI: 0.5, 29.8) and vaccination coverage (bovine respiratory syncytial virus, parainfluenza virus type 3 [33.1 pp; Cl: 15.7-50.6] and Mannheimia haemolytica [23.1 pp; Cl: 3.4-39.8]). The herds that participated in this study were likely among the more motivated regarding BRD control. As such, the information on vaccination coverage is likely not entirely representative of herds in the study area. Interpretation is further complicated by the fact that vaccinated herds were potentially less likely to face a BRD outbreak and therefore participate in the current study. This study reveals differences in the primary use of (N)SAIDs, type of antimicrobials used and vaccination coverage on beef and dairy farms in the study region, and also differences in the appropriateness of antimicrobial selection based on the Belgian formulary.

PSXIII-B-17 Risk of Bovine Respiratory Disease by Pathogen Incidence

Abstract Bovine respiratory disease (BRD) affects millions of cattle in the US annually. This study sought to identify commensal pathogens in animals without clinical signs that were predictive of clinical BRD using the Wisconsin Health Scoring (WHS) system. Cases were healthy cattle that were later diagnosed with BRD (WHS ≥ 5.0), positive controls (PC) were cattle that were diagnosed with BRD at the time of initial sampling and negative controls (NC) were those that remained healthy throughout the study (WHS ≤ 4.0). Beef feedlot cattle were sampled in Colorado (CO; cases = 13, NC = 500, PC = 488) and Washington (WA; cases = 5, NC = 499, PC = 506). Preweaned Holstein dairy calves were sampled from California (CA; cases = 73, NC = 1017, PC = 1003) and New Mexico (NM; cases = 25, NC = 381, PC = 376). For each animal, mid-nasal and deep pharyngeal swabs were used to test for nine BRD pathogens. ANOVA testing (P&amp;lt; 0.05) and odds ratios (OR) were calculated for each population. In CO, bovine viral diarrhea virus (P = 0.0001, OR = 13.1) and Histophilus somni (P = 0.008, OR = 4.2) were predictive of cattle that would be diagnosed with BRD while bovine respiratory syncytial virus (BRSV; P = 1.1 × 10-6, OR = 40.8) was most predictive in WA. In NM, BRSV (P = 0.005, OR = 5.8) was also predictive of clinical BRD. No pathogen was predictive of BRD risk in CA. Prevalence of the predictive pathogen was higher (P &amp;lt; 0.05) in cases than in PC cattle suggesting that the incidence of pathogens taken at the time of BRD diagnosis may not reflect the pathogen initiating disease. These results also highlight geographical differences in the incidence of BRD pathogens which should be considered when managing to reduce disease.

Longitudinal study of the immune response and memory following natural bovine respiratory syncytial virus infections in cattle of different age.

Human and bovine respiratory syncytial virus (HRSV and BRSV) are closely genetically related and cause respiratory disease in their respective host. Whereas HRSV vaccines are still under development, a multitude of BRSV vaccines are used to reduce clinical signs. To enable the design of vaccination protocols to entirely stop virus circulation, we aimed to investigate the duration, character and efficacy of the immune responses induced by natural infections. The systemic humoral immunity was monitored every two months during two years in 33 dairy cattle in different age cohorts following a natural BRSV outbreak, and again in selected individuals before and after a second outbreak, four years later. Local humoral and systemic cellular responses were also monitored, although less extensively. Based on clinical observations and economic losses linked to decreased milk production, the outbreaks were classified as moderate. Following the first outbreak, most but not all animals developed neutralising antibody responses, BRSV-specific IgG1, IgG2 and HRSV F- and HRSV N-reactive responses that lasted at least two years, and in some cases at least four years. In contrast, no systemic T cell responses were detected and only weak IgA responses were detected in some animals. Seronegative sentinels remained negative, inferring that no new infections occurred between the outbreaks. During the second outbreak, reinfections with clinical signs and virus shedding occurred, but the signs were milder, and the virus shedding was significantly lower than in naïve animals. Whereas the primary infection induced similar antibody titres against the prefusion and the post fusion form of the BRSV F protein, memory responses were significantly stronger against prefusion F. In conclusion, even if natural infections induce a long-lasting immunity, it would probably be necessary to boost memory responses between outbreaks, to stop the circulation of the virus and limit the potential role of previously infected adult cattle in the chain of BRSV transmission.

Reducing Calf Mortality in Ethiopia

Simple SummaryDisease and death of young livestock cause financial and production difficulties to farmers around the world. High rates of disease and death occur in various production systems in Ethiopia, hampering livestock production, reducing incomes, and damaging livelihoods. Over the last 10 years, studies carried out in Ethiopia have reported death and disease incidence rates in young livestock as high as 31% and 67%, respectively. Diarrhea and respiratory infections are the two leading causes of disease and death in calves in all production systems. In this paper, we describe findings from the experience of the Young Stock Mortality Reduction Consortium. This unique group produced important information on the main causes of disease and death in Ethiopia and created activities for small-scale farmers to address these problems. We found that several diseases caused diarrhea and respiratory infections in young calves in Ethiopia. Improving farmer knowledge and behaviors with respect to basic livestock management led to considerable reductions in young livestock disease and death and has the potential to help improve livestock productivity and human livelihoods in Ethiopia.Morbidity and mortality of young stock present economic and production challenges to livestock producers globally. In Ethiopia, calf morbidity and mortality rates, particularly due to diarrhea and respiratory disease, are high, limiting production, incomes, and the ability of farmers to improve their livelihoods. In this paper, we present findings from the combined experience of the Young Stock Mortality Reduction Consortium, which conducted epidemiological and intervention testing in calves across three production systems. This innovative alliance identified Cryptosporidium parvum and E. Coli K99 as the most common causes of diarrhea in pastoral and peri-urban calves; Strongyloides spp. as the most common fecal parasite in mixed crop–livestock and peri-urban calves; and bovine adenovirus, parainfluenza virus-3, and bovine respiratory syncytial virus as the most common respiratory pathogens in peri-urban calves. Furthermore, by improving producer knowledge with respect to fundamental livestock husbandry, feeding, housing, and neonatal care practices, calf mortality risk across production systems was reduced by 31.4 to 71.4% compared to baseline (between 10.5 and 32.1%), whereas risk of diarrhea was reduced by 52.6–75.3% (baseline between 11.4 and 30.4%) and risk of respiratory disease was reduced by 23.6–80.8% (baseline between 3.3 and 16.3%). These findings have informed scaling strategies and can potentially contribute to improved livestock productivity and human livelihoods in Ethiopia.

Clinical and microbiological effects in high-risk beef calves administered intranasal or parenteral modified-live virus vaccines.

Experimental bovine respiratory syncytial virus (BRSV) infection can enhance Histophilus somni (Hs) disease in calves; we thus hypothesized that modified-live virus (MLV) vaccines containing BRSV may alter Hs carriage. Our objective was to determine the effects of an intranasal (IN) trivalent (infectious bovine rhinotracheitis virus [IBRV], parainfluenza-3 virus [PI3V], and BRSV) respiratory vaccine with parenteral (PT) bivalent bovine viral diarrhea virus (BVDV) type I + II vaccine, or a PT pentavalent (BVDV type I and II, IBRV, BRSV, and PI3V) respiratory vaccine, on health, growth, immunity, and nasal pathogen colonization in high-risk beef calves. Calves (n = 525) were received in five truckload blocks and stratified by body weight (213 ± 18.4 kg), sex, and presence of a pre-existing ear-tag. Pens were spatially arranged in sets of three within a block and randomly assigned to treatment with an empty pen between treatment groups consisting of: 1) no MLV respiratory vaccination (CON), 2) IN trivalent MLV respiratory vaccine with PT BVDV type I + II vaccine (INT), or 3) PT pentavalent, MLV respiratory vaccine (INJ). The pen was the experimental unit, with 15 pens/treatment and 11 to 12 calves/pen in this 70-d receiving study. Health, performance, and BRSV, Hs, Mycoplasma bovis (Mb), Mannheimia haemolytica (Mh), and Pasteurella multocida (Pm) level in nasal swabs via rtPCR was determined on days 0, 7, 14, and 28, and BRSV-specific serum neutralizing antibody titer, and serum IFN-γ concentration via ELISA, were evaluated on days 0, 14, 28, 42, 56, and 70. Morbidity (P = 0.83), mortality (P = 0.68) and average daily gain (P ≥ 0.82) did not differ. Serum antibodies against BRSV increased with time (P < 0.01). There was a treatment × time interaction (P < 0.01) for Hs detection; on days 14 and 28, INT (21.1% and 57.1%) were more frequently (P < 0.01) Hs positive than CON (3.6% and 25.3%) or INJ (3.4 % and 8.4%). Also, INT had reduced (P = 0.03) cycle time of Hs positive samples on day 28. No difference (P ≥ 0.17) was found for IFN-γ concentration and Mb, Mh, or Pm detection. The proportion of Mh positive culture from lung specimens differed (P < 0.01); INT had fewer (0.0%; 0 of 9) Mh positive lungs than INJ (45.5%; 6 of 13) or CON (74.0%; 14 of 19). Vaccination of high-risk calves with MLV did not clearly impact health or growth during the receiving period. However, INT was associated with an altered upper respiratory microbial community in cattle resulting in increased detection and level of Hs.

Mycoplasma bovis is associated with Mannheimia haemolytica during acute bovine respiratory disease in feedlot cattle.

Bovine Respiratory Disease (BRD) represents a significant burden to the health of feedlot cattle and the profitability of the beef industry in the US. Mannheimia haemolytica is widely regarded as the primary bacterial pathogen driving acute BRD. While Mycoplasma bovis is most commonly implicated in chronic cases of BRD, this agent's potential role in acute stages of BRD is unclear. Therefore, this study aimed to evaluate potential associations between M. bovis and M. haemolytica during acute BRD in feedlot cattle. Nasal swabs (n = 1,044) were collected over time from feedlot cattle (n = 270) enrolled in an experiment assessing the effect of vaccination for Bovine Respiratory Syncytial Virus (BRSV). Swabs were analyzed for detection of M. bovis, M. haemolytica, Pasteurella multocida, Histophilus somni, and BRSV via multiplex qPCR assays. Data were analyzed using inverse conditional probability weighted (ICPW) logistic regression models to investigate potential effects of M. bovis presence on arrival (d0), day seven (d7) and day 14 (d14) post-arrival on M. haemolytica prevalence on day 28 (d28) post-arrival, adjusting for the previous history of P. multocida, H. somni, BRSV, BRD morbidity, and body weight. The potential association between time-to-BRD detection and M. bovis presence on d0, d7, and d14 post-arrival, was inferred via an ICPW time-to-event model. The presence of M. bovis in nasal swabs collected on d7 post-arrival was significantly associated with an increase in the prevalence of M. haemolytica on d28 (prevalence difference: 45%; 95% Confidence Interval: 31%, 60%; P-value < 0.001). Significant time-varying coefficients for M. bovis presence were detected at d0, d7, and d14 post-arrival in the ICPW time-to-event model (P-value < 0.001). The shortest median time-to-BRD detection was 29 days in cattle that were M. bovis positive on d0, d7, and d14 post-arrival and in those that were positive on d0 and d14 post-arrival. Under the conditions of this study, our findings suggest that M. bovis may be influencing the respiratory environment during the acute phase of BRD, increasing the abundance of M. haemolytica, which could have important impacts on the occurrence of BRD.

The Prevalence, Coexistence, and Correlations between Seven Pathogens Detected by a PCR Method from South-Western Poland Dairy Cattle Suffering from Bovine Respiratory Disease.

Bovine respiratory disease (BRD) is a very important disease that contributes to economic losses in dairy and beef cattle breeding worldwide. The molecular testing of material from 296 calves showing BRD symptoms from 74 dairy herds located in south-western Poland was performed in 2019–2021. Molecular tests were performed using a commercial kit “VetMAXTM Ruminant Respiratory Screening Kit” (Thermo Fisher Scientific) for the simultaneous detection of genetic material of seven pathogens responsible for BRD. At least one pathogen was detected in 95.95% of herds. The overall prevalence was: Pasteurella multocida 87.84%, Mannheimia haemolytica 44.59%, bovine coronavirus (BcoV) 32.43%, Mycoplasma bovis 29.73%, Histophilus somni 28.38%, bovine parainfluenza virus type 3 (BPIV-3) 13.51%, and bovine respiratory syncytial virus (BRSV) 10.81%. Twenty-nine configurations of pathogen occurrences were found. Bacterial infections were the most frequently recorded as 56.7% of all results. Coinfections mainly consisted of two pathogens. Not a single purely viral coinfection was detected. The most frequent result was a single P. multocida infection accounting for 18.31% of all results. The statistically significant correlation (p = 0.001) with the highest strength of effect (ϕ 0.38) was between M. bovis and H. somni.

Individual antimicrobial treatments in veal calves: Effect on the net carcasses weight at the slaughterhouse and relationship with the serostatus of the calves upon arrival to the fattening unit

In recent years, the scientific community, institutions, and public opinion have shown a growing awareness towards the problem of antimicrobial resistance. Guidelines from the European Union aim at significantly reducing (even avoiding) the antimicrobial use for prophylactic and metaphylactic purposes in veterinary medicine and this represents an important issue for several intensive farming systems, such as the veal industry. This retrospective observational cohort study analyzed net carcass weight data at the slaughter of 618 veal calves belonging to 6 batches of animals fattened in an Italian commercial farm according to the number of individual antimicrobial treatments administered during the fattening cycle. Sixty-eight percent (419 animals) of the overall sample of calves received at least one individual antimicrobial treatment due to a disease event during the fattening, and 280 of these (45 % of the overall sample) had more than one treatment. On average, the net carcass weight of calves individually treated more than once was about 10 kg lower than that of untreated calves or treated only once. Moreover, the carcasses of calves that received more than one treatment were 16 times more at risk of being severely penalized in payment at the slaughterhouse due to a net weight below 110 kg than those of untreated calves. Serum analysis data of a subsample of 105 calves belonging to 3 out of the 6 batches collected within 5 days after their arrival to the veal farm identified 3 persistently infected animals by bovine viral diarrhea (BVD) virus and a prevalence of calves with serum immunoglobulin concentration below 7.5 g/L of 37 %. The presence of specific antibodies against infectious bovine rhinotracheitis (IBR) and bovine respiratory syncytial virus (BRSV) and a serum immunoglobulin level above 7.5 g/L at the onset of the fattening were the most important factors associated with a decreased risk of multiple individual antimicrobial treatments (and thus, disease development) or with a delay in disease onset during the fattening, thus leading to better performances at the slaughter. The outcomes of this study suggested that a relationship could exist between calf serostatus upon arrival to the veal farm and the possibility to avoid multiple individual antimicrobial treatments during fattening. Further research is needed to deeply study this association, as such knowledge would allow for planning effective preventing strategies to reduce the antimicrobial use in veal calves.

Infectious Disease Agents Associated with Pulmonary Alterations in Aborted Bovine Fetuses.

Simple SummaryA retrospective study was performed to identify pulmonary alterations and/or pneumonia in aborted bovine fetuses (n = 37) and to associate the presence of infectious disease antigens and nucleic agents with patterns and/or alterations of pulmonary disease. Immunohistochemical (IHC) assays were performed to identify antigens of selected agents associated with bovine respiratory disease: bovine alphaherpesvirus 1 (BoAHV1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV-3), bovine respiratory syncytial virus (BRSV), and Mycoplasma bovis. Molecular assays were performed to identify nucleic acids of a panel of reproductive disease agents and bovine gammaherpesvirus 6 (BoGHV6) in the lungs of 12 fetuses. Only interstitial pneumonia (12/37) and suppurative bronchopneumonia (1/37) were observed; pneumonia was not observed in 65% of the tissues evaluated. The most frequent intralesional agents were BRSV (16.2%; 6/37), BVDV (13.5%; 5/37), and BoAHV1 (8.1%; 3/37). Interstitial pneumonia was associated with BRSV (n = 3), BoAHV1 (n = 3), and BVDV (n = 2); suppurative bronchopneumonia contained a Gram-positive bacterium and intralesional antigens of BVDV and BRSV. Nucleic acid detection identified at least one disease agent in 75% of the fetuses examined. Reproductive pathogens detected included Leptospira spp., (n = 3), BVDV, Neospora caninum, and Brucella abortus (n = 2). BoGHV6 DNA was identified in the lungs of two fetuses with interstitial pneumonia. Single (n = 7), dual (n = 3), triple (n = 4), and quadruple (n = 1) mixed infections were identified while infectious agents were not identified in 59.5% (22/37) of the examined lungs. Single fetal infections were associated with BRSV, BVDV (n = 2), Leptospira spp., BoAHV1, and BoGHV6 (n = 1). These results indicate that the fetuses suffered intrauterine infection through transplacental transmission. The identification of BRSV and BoGHV6 in multiple fetuses with associated pulmonary alterations warrants further investigation relative to the role of these agents in fetopathy and possible direct and/or indirect effects on fetal survival.This study investigated the occurrence of selected pathogens of bovine respiratory disease in fetal pulmonary tissue of cattle and associated these with patterns of disease. Fetal pulmonary (n = 37) tissues were evaluated by histopathology; immunohistochemical assays identified intralesional antigens of bovine alphaherpesvirus 1 (BoAHV1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV-3), bovine respiratory syncytial virus (BRSV), and Mycoplasma bovis. Molecular assays were performed to amplify reproductive disease pathogens and bovine gammaherpesvirus 6 (BoGHV6) from 12 lungs. The 2 patterns of pulmonary diseases were interstitial pneumonia (12/37) and suppurative bronchopneumonia (1/37). The frequency of the intralesional antigens identified was BRSV (16.2%; 6/37), BVDV (13.5%; 5/37), BoAHV1 (8.1%; 3/37), M. bovis (5.4%; 2/37), and BPIV-3 (2.7%; 1/37). Interstitial pneumonia was associated with BRSV (n = 3), BoAHV1 (n = 3), and BVDV (n = 2); suppurative bronchopneumonia contained a Gram-positive bacterium and BVDV and BRSV. Reproductive pathogens detected included Leptospira spp., (n = 3), BVDV, Neospora caninum, and Brucella abortus (n = 2). BoGHV6 DNA was identified in the lungs of two fetuses with interstitial pneumonia. These findings suggest that these fetuses were infected transplacentally by several pathogens. The role of some of these pathogens herein identified must be further elucidated in the possible participation of fetal disease.

Bovine Coronavirus Co-infection and Molecular Characterization in Dairy Calves With or Without Clinical Respiratory Disease.

Bovine respiratory disease (BRD) is considered a major cause of morbidity and mortality in young calves and is caused by a range of infectious agents, including viruses and bacteria. This study aimed to determine the frequency of viral and bacterial pathogens detected in calves with BRD from high-production dairy cattle herds and to perform the molecular characterization of N and S1 genes in identified bovine coronavirus (BCoV) strains. Nasal swabs were collected from 166 heifer calves, namely, 85 symptomatic and 81 asymptomatic calves aged between 5 and 90 days, from 10 dairy cattle herds. Nasal swabs were evaluated using molecular techniques for the identification of viruses (BCoV, bovine alphaherpesvirus 1, bovine viral diarrhea virus, bovine parainfluenza virus 3, and bovine respiratory syncytial virus) and bacteria (Pasteurella multocida, Mannheimia haemolytica, Histophilus somni, and Mycoplasma bovis). In addition, five and two BCoV-positive samples were submitted to N and S1 gene amplification and nucleotide sequencing, respectively. The frequency of diagnosis of BCoV was higher (56%, 93/166) than the frequency of P. multocida (39.8%, 66/166) and M. haemolytica (33.1%, 55/166). The three microorganisms were identified in the calves of symptomatic and asymptomatic heifer calve groups. All other pathogens included in the analyses were negative. In the phylogenetic analysis of the S1 gene, the Brazilian strains formed a new branch, suggesting a new genotype, called # 15; from the N gene, the strains identified here belonged to cluster II. This study describes high rates of BCoV, P. multocida, and M. haemolytica in heifer calves from high-production dairy cattle herds with BRD. Additionally, the molecular characterization provides evidence that the circulating BCoV strains are ancestrally different from the prototype vaccine strains and even different BCoV strains previously described in Brazil.

Characterization of tropical cattle production units: Effect of sanitary management

Objective: To characterize dual-purpose cattle production units (PUs) and to identify the sanitary management that producers use with their herds. Design/Methodology/Approach: Sixty (60) cattle producers were interviewed; the questions were related to the characteristics of the PU and the sanitary status. Four serum samples from the largest herd were sent to the laboratory to detect diseases. Results: All the cattle producers surveyed were men between 20 and 50 years old. Of them, 40% have been devoted to the activity for 11 to 25 years. The PUs have on average 25 animals, 45% of producers have between 21 and 30 cows, and the highest percentage of animals are breeds Gyr (35%) and Sardo Negro (30%). Of producers, 65% tend to their sick animals. On the other hand, the laboratory studies showed that three cows were positive for Bovine Respiratory Syncytial Virus (BRSV), two cows were positive for type 1 bovine herpes virus (BoHV-1), or Infectious Bovine Rhinotracheitis (IBR) and parainfluenza 3 (PI3), and finally all the cows were positive for Leptospira sp. Hardjo. Study Limitations/Implications: Under the conditions in which the study was conducted, it is costly to send samples to the laboratory. The producer with the largest herd financed the laboratory tests of the cows sampled. This information is useful for local producers with regards to the characteristics in which they produce them. Findings/Conclusions: Dual-purpose cattle producers in the tropics produce at a small scale; and one of the largest PUs presents problems of abortive diseases.

Immunization with a mucosal, post-fusion F/G protein-based polyanhydride nanovaccine protects against BRSV infection in neonatal calves

Abstract Human respiratory syncytial virus (HRSV) is a leading cause of severe acute lower respiratory tract (LRT) disease in young children worldwide. Currently, there is no approved vaccine for HRSV. Bovine RSV (BRSV) is closely related to HRSV and is the cause of severe acute LRT disease in young cattle. A neonatal calf model presents an opportunity to test HRSV vaccines and study a naturally susceptible host-pathogen interaction similar to HRSV infection. We tested the efficacy of a polyanhydride nanoparticle-vaccine (nanovaccine) encapsulating the post-fusion F and G proteins from BRSV and CpG adjuvant in neonatal calves with maternally-derived antibodies (MDA). A prime-boost immunization of neonatal calves with the BRSV post-fusion F/G CpG nanovaccine induced clinical protection in the presence of MDA. Vaccinated calves developed moderate or no clinical symptoms and were mostly protected from virus-associated lung pathology compared to unvaccinated controls. Calves that received a mucosal/systemic nanovaccine immunization had complete virological protection against BRSV infection in the lungs, which correlated with BRSV-specific immune responses in the bronchoalveolar lavage. Vaccination with the nanovaccine induced anamnestic BRSV-specific IgA titers in the lungs and systemic T cell responses with BRSV-specific CD4+ T cell proliferation and antigen-specific IL-17 and IFNγ secretion. Overall, the BRSV post-fusion F/G CpG nanovaccine is a promising candidate for further studies to protect neonatal calves and infants from RSV infection.

Altered transcriptome responses in the lungs of preweaned calves supplemented with a yeast fermentate during a viral-bacterial coinfection

Abstract Bovine respiratory disease is a complex syndrome which contributes to severe and often fatal pneumonia in beef and dairy cattle. Saccharomyces cerevisiae fermentation products (SCFP) are feed ingredients that have been previously shown to alter the immune response and outcome of a respiratory viral infection in calves. The objective of this study was to determine the effect of SCFP supplementation on the lung transcriptome in calves responding to a co-infection with bovine respiratory syncytial virus (BRSV) and Pasteurella multocida (PM). Twenty-eight, 1–2 day old calves were assigned to two groups: 1) control diet; or 2) SCFP treated diet. Calves were infected with BRSV on day 21, followed 6 days later by intratracheal inoculation with PM. Calves were euthanized on day 10 post-viral infection. There were no differences in viral or bacterial burden between treatment groups, but SCFP treated calves tended to have less severe lung pathology. Transcriptome analysis of lung tissue and bronchoalveolar lavage samples revealed over-represented genes related to membrane integrity, pathogen binding, and complement activation which presumably contributes to the host’s resiliency and response to the coinfection. Results from this study suggest that supplementing preweaned calves with SCFP may modulate the innate and adaptive immune response in mucosal sites. These findings will contribute to a better understanding of the underlying host response to bovine respiratory disease. Supported by funding from Diamond V Mills, Inc., Cedar Rapids, IA

Bovine Respiratory Syncytial Virus Genome Sequences from Cattle with Clinical Respiratory Disease in Kansas, 2021.

ABSTRACTWe report two near-complete bovine respiratory syncytial virus genome sequences collected from 10-month-old cattle with respiratory disease in Kansas in December 2021. No other respiratory pathogens were confirmed in the samples. These genome sequences update the currently circulating BRSV field strains in the United States.

Comparative Histopathology of Bovine Acute Interstitial Pneumonia and Bovine Respiratory Syncytial Virus-Associated Interstitial Pneumonia

Acute interstitial pneumonia (AIP) is a significant disease of cattle and many aetiologies have been implicated on the basis of the characteristic pathological lesions. Bovine respiratory syncytial virus (BRSV) is one of the key aetiological factors in bovine respiratory disease complex and several studies have suggested, controversially, that BRSV may be an underlying cause of bovine AIP. BRSV infection is known to cause several distinctive histopathological changes, including epithelial syncytia formation and intracytoplasmic viral inclusions. However, distinguishing bovine AIP from BRSV-related pneumonia by clinical presentation, gross pathology or histopathology can sometimes be challenging. In order to identify the potential distinguishing features, we compared the histopathological findings of AIP that were, and were not, associated with BRSV infection in naturally occurring cases. We found that multinucleated giant cells were more frequently identified in cattle with AIP while bronchiolitis was more common in BRSV-infected cattle. However, this was not considered a sole indicator of either disease group. Statistically, we identified that a combination of several histopathological features, including alveolar septal necrosis, presence of multinucleated giant cells and bronchiolitis, can serve as an excellent indicator for distinguishing between idiopathic AIP and BRSV-related pneumonia, with a strong statistical significance (P=0.0004). Based on the results of this retrospective study, we present a histopathological scoring system for predicting BRSV-associated AIP.

Limitations of bacterial culture, viral PCR, and tulathromycin susceptibility from upper respiratory tract samples in predicting clinical outcome of tulathromycin control or treatment of bovine respiratory disease in high-risk feeder heifers.

A cross-sectional prospective cohort study including 1026 heifers administered tulathromycin due to high risk of clinical signs of bovine respiratory disease (BRD), measured poor association between BRD clinical outcomes and results of bacterial culture and tulathromycin susceptibility from BRD isolates of deep nasopharyngeal swabs (DNS) and adequate association with viral polymerase chain reaction (PCR) results from nasal swabs. Isolation rates from DNS collected on day-0 and at 1st BRD-treatment respectively were: Mannheimia haemolytica (10.9% & 34.1%); Pasteurella multocida (10.4% & 7.4%); Mycoplasma bovis (1.0% & 36.6%); and Histophilus somni (0.7% & 6.3%). Prevalence of BRD viral nucleic acid on nasal swabs collected exclusively at 1st BRD-treatment were: bovine parainfluenza virus type-3 (bPIV-3) 34.1%; bovine viral diarrhea virus (BVDV) 26.3%; bovine herpes virus type-1 (BHV-1) 10.8%; and bovine respiratory syncytial virus (BRSV) 54.1%. Increased relative risk, at 95% confidence intervals, of 1st BRD-treatment failure was associated with positive viral PCR results: BVDV 1.39 (1.17–1.66), bPIV-3 1.26 (1.06–1.51), BHV-1 1.52 (1.25–1.83), and BRSV 1.35 (1.11–1.63) from nasal swabs collected at 1st BRD-treatment and culture of M. haemolytica 1.23 (1.00–1.51) from DNS collected at day-0. However, in this population of high-risk feeder heifers, the predictive values of susceptible and resistant isolates had inadequate association with BRD clinical outcome. These results indicate, that using tulathromycin susceptibility testing of isolates of M. haemolytica or P. multocida from DNS collected on arrival or at 1st BRD-treatment to evaluate tulathromycin clinical efficacy, is unreliable.

Development of a One-Step Multiplex Real-Time PCR Assay for the Detection of Viral Pathogens Associated With the Bovine Respiratory Disease Complex.

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.

Bovine Respiratory Syncytial Virus (BRSV) Infection Detected in Exhaled Breath Condensate of Dairy Calves by Near-Infrared Aquaphotomics.

Bovine respiratory syncytial virus (BRSV) is a major contributor to respiratory disease in cattle worldwide. Traditionally, BRSV infection is detected based on non-specific clinical signs, followed by reverse transcriptase-polymerase chain reaction (RT-PCR), the results of which can take days to obtain. Near-infrared aquaphotomics evaluation based on biochemical information from biofluids has the potential to support the rapid identification of BRSV infection in the field. This study evaluated NIR spectra (n = 240) of exhaled breath condensate (EBC) from dairy calves (n = 5) undergoing a controlled infection with BRSV. Changes in the organization of the aqueous phase of EBC during the baseline (pre-infection) and infected (post-infection and clinically abnormal) stages were found in the WAMACS (water matrix coordinates) C1, C5, C9, and C11, likely associated with volatile and non-volatile compounds in EBC. The discrimination of these chemical profiles by PCA-LDA models differentiated samples collected during the baseline and infected stages with an accuracy, sensitivity, and specificity >93% in both the calibration and validation. Thus, biochemical changes occurring during BRSV infection can be detected and evaluated with NIR-aquaphotomics in EBC. These findings form the foundation for developing an innovative, non-invasive, and in-field diagnostic tool to identify BRSV infection in cattle.

Effect of fish oil and canola oil supplementation on immunological parameters, feed intake, and growth of Holstein calves

Supplemental n-3 fatty acids (FA) may support better immune responses than n-6 and n-9 FA in dairy calves. The objective was to evaluate the effect of n-3 FA, supplemented as a fish oil product (FO) in the milk replacer (MR), in comparison to n-6 and n-9 FA, supplemented as canola oil (CO), on body weight (BW), daily gain, and immunological parameters of preweaning Holstein calves. The study was conducted from September to December 2019. Calves were randomly allocated to a control group (n = 15; BW = 36.2 ± 1.5 kg; mean ± SEM) supplemented daily with 30 mL of CO and to an experimental group (n = 15; BW = 36.3 ± 1.5 kg) supplemented with 60 g of a product containing 30 g of FO. Both treatments were added to the MR during the morning feeding. All calves were fed 4 L of MR at 12.5% solids at 0700 and 1600 h for wk 1, 6 L from wk 2 to 7, and 3 L once daily (0700 h) during wk 8 until weaning (56 d). Blood samples were collected at 7, 14, 21, 28, 35, 42, 49, and 56 d of age for serum haptoglobin, TNF-α, IL-1β, and protectin. Dry matter intake was recorded in all experimental calves daily. Seroneutralization titers to vaccination against viral diseases (infectious bovine rhinotracheitis, parainfluenza 3, bovine viral diarrhea, and bovine respiratory syncytial virus) were determined. Mixed models for repeated measures were developed to analyze variables over time. Seroneutralization titers were analyzed by the Kruskal-Wallis test. The other variables were compared by a generalized linear model. Serum FA profile at 35 d of age showed that FO supported higher concentrations of n-3 FA than CO. Final BW [65.2 vs. 62.0 kg, standard error of the mean (SEM) = 2.1 kg] and average daily gain (0.52 vs. 0.46 kg/d, SEM = 0.1 kg/d) tended to be higher for the FO than the CO group. An interaction of treatment × day for dry matter intake was observed, especially during weaning (2.17 kg vs. 1.94 kg, SEM = 0.158 kg, for FO and CO group, respectively). Blood lactate (mmol/L) was higher in the CO than in the FO group at d 7. Haptoglobin and IL-1β were higher for the CO group on d 14 than the FO group. The TNF- α concentrations for the FO group were reduced over time, whereas the concentrations in the CO group remained constant. Protectin was higher in the FO group on d 14, but was lower on d 28, 35, and 49. Seroneutralization antibody titers postvaccination for the PI3 virus were higher for the FO than the CO group. In conclusion, calves supplemented with FO had lower concentrations of blood lactate, haptoglobin, IL-1β and TNF-α than calves supplemented with CO during the study period. The FO supplementation had a higher DMI than CO supplementation. Results of this trial should be interpreted with caution due to the lack of a negative control group as well as the lower birth weight and growth rate observed under heat stress conditions.

Early Activation of the Innate Immunity and Specific Cellular Immune Pathways after Vaccination with a Live Intranasal Viral Vaccine and Challenge with Bovine Parainfluenza Type 3 Virus.

Bovine parainfluenza type 3 (BPIV3) and bovine respiratory syncytial virus (BRSV) may cause bovine respiratory disease (BRD) in very young calves, and therefore vaccination should induce protection at the youngest age and as quickly as possible. This can be achieved by intranasal vaccination with a vaccine containing live attenuated BRSV and BPIV3 virus strains. The objective of this study was to measure gene expression levels by means of RT-qPCR of proteins involved in the innate and adaptive immune response in the nasopharyngeal mucosae after administration of the above-mentioned vaccine and after challenge with BPIV3. Gene expression profiles were different between (i) vaccinated, (ii) nonvaccinated-challenged, and (iii) vaccinated-challenged animals. In nonvaccinated-challenged animals, expression of genes involved in development of disease symptoms and pathology were increased, however, this was not the case after vaccination. Moreover, gene expression patterns of vaccinated animals reflected induction of the antiviral and innate immune pathways as well as an initial Th1 (cytotoxic) cellular response. After challenge with BPIV3, the vaccinated animals were protected against nasal shedding of the challenge virus and clinical symptoms, and in parallel the expression levels of the investigated genes had returned to values that were found before vaccination. In conclusion, in comparison to the virulent wild-type field isolates, the two virus strains in the vaccine have lost their capacity to evade the immune response, resulting in the induction of an antiviral state followed by a very early activation of innate immune and antiviral responses as well as induction of specific cellular immune pathways, resulting in protection. The exact changes in the genomes of these vaccine strains leading to attenuation have not been identified. These data represent the real-life situation and can serve as a basis for further detailed research. This is the first report describing the effects on immune gene expression profiles in the nasal mucosae induced by intranasal vaccination with a bivalent, live BRSV-BPI3V vaccine formulation in comparison to wild-type infection with a virulent BPI3V strain.