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.

Enhanced Pathogenesis Caused by Influenza D Virus and Mycoplasma bovis Coinfection in Calves: a Disease Severity Linked with Overexpression of IFN-γ as a Key Player of the Enhanced Innate Immune Response in Lungs.

Bovine respiratory disease (BRD) is a major disease of young cattle whose etiology lies in complex interactions between pathogens and environmental and host factors. Despite a high frequency of codetection of respiratory pathogens in BRD, data on the molecular mechanisms and pathogenesis associated with viral and bacterial interactions are still limited. In this study, we investigated the effects of a coinfection with influenza D virus (IDV) and Mycoplasma bovis in cattle. Naive calves were infected by aerosol with a French IDV strain and an M. bovis strain. The combined infection shortened the incubation period, worsened the disease, and led to more severe macroscopic and microscopic lesions compared to these parameters in calves infected with only one pathogen. In addition, IDV promoted colonization of the lower respiratory tract (LRT) by M. bovis and increased white cell recruitment to the airway lumen. The transcriptomic analysis highlighted an upregulation of immune genes in the lungs of coinfected calves. The gamma interferon (IFN-γ) gene was shown to be the gene most statistically overexpressed after coinfection at 2 days postinfection (dpi) and at least until 7 dpi, which correlated with the high level of lymphocytes in the LRT. Downregulation of the PACE4 and TMPRSS2 endoprotease genes was also highlighted, being a possible reason for the faster clearance of IDV in the lungs of coinfected animals. Taken together, our coinfection model with two respiratory pathogens that when present alone induce moderate clinical signs of disease was shown to increase the severity of the disease in young cattle and a strong transcriptomic innate immune response in the LRT, especially for IFN-γ. IMPORTANCE Bovine respiratory disease (BRD) is among the most prevalent diseases in young cattle. BRD is due to complex interactions between viruses and/or bacteria, most of which have a moderate individual pathogenicity. In this study, we showed that coinfection with influenza D virus (IDV) and Mycoplasma bovis increased the severity of the respiratory disease in calves in comparison with IDV or M. bovis infection. IDV promoted M. bovis colonization of the lower respiratory tract and increased white cell recruitment to the airway lumen. The transcriptomic analysis highlighted an upregulation of immune genes in the lungs of coinfected calves. The IFN-γ gene in particular was highly overexpressed after coinfection, correlated with the disease severity, immune response, and white cell recruitment in the lungs. In conclusion, we showed that IDV facilitates coinfections within the BRD complex by modulating the local innate immune response, providing new insights into the mechanisms involved in severe respiratory diseases.

Haptoglobin and its association with naturally occurring diseases in Holstein heifer calves / [Haptoglobina e sua associação com doenças ocorridas naturalmente em bezerras da raça Holandesa]

The present study evaluated the use of haptoglobin (Hp) as an indicator of health and performance in 166 Holstein heifer calves reared in an intensive production system. Calves were evaluated at D6-9; D10-13; D20-23; D35-38 and D65-68, corresponding to the days of life. The absence or presence of diseases was evaluated by physical examination and classification of scores. The performance parameters evaluated were body weight, height at withers and hind width. Hp was measured by spectrophotometric technique. The highest prevalence of diarrhea (59.4%; 98/165) was observed in D10-13, bovine respiratory disease (BRD) was on D35-38 (25.8%; 42/163), and umbilical inflammations in D6-D9 (7.8%; 13/166). Highest values of Hp were observed in animals with diarrhea (P=0.02), and umbilical inflammation (P=0.057), in comparison with the group of healthy calves. A significant negative correlation was observed between Hp and performance index. This protein presented an important relation with diarrhea and performance of the calves, opening perspectives on its utilization as a biomarker of diseases.(AU)

O presente estudo avaliou o uso da haptoglobina (Hp) como indicadora de sanidade e desempenho em 166 bezerras Holandesas criadas em um sistema de produção intensivo. As bezerras foram avaliadas nos momentos D6-9; D10-13; D20-23; D35-38 e D65-68, sendo estes correspondentes aos dias de vida. A ausência ou a presença de doenças foi avaliada por meio do exame físico e da classificação por escores. Os parâmetros de desempenho avaliados foram peso corporal, altura de cernelha e largura de garupa. A Hp foi mensurada por técnica espectrofotométrica. A maior prevalência de diarreia (59,4%; 98/165) foi observada em D10-13, doença respiratória bovina (DRB) ocorreu em D35-38 (25,8%; 42/163) e inflamações umbilicais em D6-D9 (7,8%; 13/166). O valor de Hp foi maior nos animais que apresentaram diarreia (P=0,02) e inflamações umbilicais (P=0,057), em comparação ao grupo de bezerras saudáveis. Houve correlação negativa significativa entre a Hp e os índices de desempenho. Essa proteína apresentou uma importante relação com a diarreia e com o desempenho das bezerras, abrindo perspectivas sobre a sua utilização como biomarcadora de doenças.(AU)

Comprehensive at-arrival transcriptomic analysis of post-weaned beef cattle uncovers type I interferon and antiviral mechanisms associated with bovine respiratory disease mortality.

BACKGROUND: Despite decades of extensive research, bovine respiratory disease (BRD) remains the most devastating disease in beef cattle production. Establishing a clinical diagnosis often relies upon visual detection of non-specific signs, leading to low diagnostic accuracy. Thus, post-weaned beef cattle are often metaphylactically administered antimicrobials at facility arrival, which poses concerns regarding antimicrobial stewardship and resistance. Additionally, there is a lack of high-quality research that addresses the gene-by-environment interactions that underlie why some cattle that develop BRD die while others survive. Therefore, it is necessary to decipher the underlying host genomic factors associated with BRD mortality versus survival to help determine BRD risk and severity. Using transcriptomic analysis of at-arrival whole blood samples from cattle that died of BRD, as compared to those that developed signs of BRD but lived (n = 3 DEAD, n = 3 ALIVE), we identified differentially expressed genes (DEGs) and associated pathways in cattle that died of BRD. Additionally, we evaluated unmapped reads, which are often overlooked within transcriptomic experiments. RESULTS: 69 DEGs (FDR<0.10) were identified between ALIVE and DEAD cohorts. Several DEGs possess immunological and proinflammatory function and associations with TLR4 and IL6. Biological processes, pathways, and disease phenotype associations related to type-I interferon production and antiviral defense were enriched in DEAD cattle at arrival. Unmapped reads aligned primarily to various ungulate assemblies, but failed to align to viral assemblies. CONCLUSION: This study further revealed increased proinflammatory immunological mechanisms in cattle that develop BRD. DEGs upregulated in DEAD cattle were predominantly involved in innate immune pathways typically associated with antiviral defense, although no viral genes were identified within unmapped reads. Our findings provide genomic targets for further analysis in cattle at highest risk of BRD, suggesting that mechanisms related to type I interferons and antiviral defense may be indicative of viral respiratory disease at arrival and contribute to eventual BRD mortality.

Predicting bovine respiratory disease outcome in feedlot cattle using latent class analysis.

Bovine respiratory disease (BRD) is the most significant disease affecting feedlot cattle. Indicators of BRD often used in feedlots such as visual signs, rectal temperature, computer-assisted lung auscultation (CALA) score, the number of BRD treatments, presence of viral pathogens, viral seroconversion, and lung damage at slaughter vary in their ability to predict an animal's BRD outcome, and no studies have been published determining how a combination of these BRD indicators may define the number of BRD disease outcome groups. The objectives of the current study were (1) to identify BRD outcome groups using BRD indicators collected during the feeding phase and at slaughter through latent class analysis (LCA) and (2) to determine the importance of these BRD indicators to predict disease outcome. Animals with BRD (n = 127) were identified by visual signs and removed from production pens for further examination. Control animals displaying no visual signs of BRD (n = 143) were also removed and examined. Blood, nasal swab samples, and clinical measurements were collected. Lung and pleural lesions indicative of BRD were scored at slaughter. LCA was applied to identify possible outcome groups. Three latent classes were identified in the best model fit, categorized as non-BRD, mild BRD, and severe BRD. Animals in the mild BRD group had a higher probability of having visual signs of BRD compared with non-BRD and severe BRD animals. Animals in the severe BRD group were more likely to require more than 1 treatment for BRD and have ≥40 °C rectal temperature, ≥10% total lung consolidation, and severe pleural lesions at slaughter. Animals in the severe BRD group were also more likely to be naïve at feedlot entry and the first BRD pull for Bovine Viral Diarrhoea Virus, Bovine Parainfluenza 3 Virus, and Bovine Adenovirus and have a positive nasal swab result for Bovine Herpesvirus Type 1 and Bovine Coronavirus. Animals with severe BRD had 0.9 and 0.6 kg/d lower overall ADG (average daily gain) compared with non-BRD animals and mild BRD animals (P < 0.001). These results demonstrate that there are important indicators of BRD severity. Using this information to predict an animal's BRD outcome would greatly enhance treatment efficacy and aid in better management of animals at risk of suffering from severe BRD.

Non-specific protection from respiratory tract infections in cattle generated by intranasal administration of an innate immune stimulant.

Alternatives to antibiotics for prevention of respiratory tract infections in cattle are urgently needed given the increasing public and regulatory pressure to reduce overall antibiotic usage. Activation of local innate immune defenses in the upper respiratory tract is one strategy to induce non-specific protection against infection with the diverse array of viral and bacterial pathogens associated with bovine respiratory disease complex (BRDC), while avoiding the use of antibiotics. Our prior studies in rodent models demonstrated that intranasal administration of liposome-TLR complexes (LTC) as a non-specific immune stimulant generated high levels of protection against lethal bacterial and viral pathogens. Therefore, we conducted studies to assess LTC induction of local immune responses and protective immunity to BRDC in cattle. In vitro, LTC were shown to activate peripheral blood mononuclear cells in cattle, which was associated with secretion of INFγ and IL-6. Macrophage activation with LTC triggered intracellular killing of Mannheimia hemolytica and several other bacterial pathogens. In studies in cattle, intranasal administration of LTC demonstrated dose-dependent activation of local innate immune responses in the nasopharynx, including recruitment of monocytes and prolonged upregulation (at least 2 weeks) of innate immune cytokine gene expression by nasopharyngeal mucosal cells. In a BRDC challenge study, intranasal administration of LTC prior to pathogen exposure resulted in significant reduction in both clinical signs of infection and disease-associated euthanasia rates. These findings indicate that intranasal administration of a non-specific innate immune stimulant can be an effective method of rapidly generating generalized protection from mixed viral and bacterial respiratory tract infections in cattle.

Associations of barn air quality parameters with ultrasonographic lung lesions, airway inflammation and infection in group-housed calves.

Barn climate is believed to play a major role in the bovine respiratory disease complex. However, the exact air quality parameters associated with (sub)clinical pneumonia or airway inflammation in calves are currently unknown. The objective of this cross-sectional study was to assess associations of air quality parameters with clinical signs, lung consolidation, pulmonary inflammation and infection in group-housed calves. In total, 60 beef and dairy farms were visited from January to April 2017 and 428 calves sampled. Measured air quality parameters included continuous 24-h measurements of ammonia concentration, relative humidity and temperature and punctual measurements of air velocity, ammonia, CO2 and bacterial air load. Calf sampling consisted of clinical examination, thoracic ultrasonography and broncho-alveolar lavage sampling for bacteriological and cytological analysis of broncho-alveolar lavage fluid (BALf). Average air temperature was 14.2 °C (standard deviation (SD) 4.4, range 5.5-23.9) and relative humidity 68.8 % (SD 8.9, range 52.2-91.6). Average ammonia concentration was 1.7 ppm (SD 0.9, range 0-10.0). Lung consolidations of ≥1 cm, ≥3 cm and ≥6 cm in depth were present in 41.1 % (176/428), 27.1 % (116/428) and 16.1 % (69/428) of the calves, respectively. Average pen temperature was positively associated with consolidations of ≥1 cm (P = 0.005), ≥3 cm (P = 0.002) and ≥6 cm (P < 0.01). Ammonia exposure, in hours>4 ppm, was associated with lung consolidation ≥1 cm (odds ratio (OR) = 1.73; confidence interval (CI) = 1.02-3.07; P = 0.04). Ammonia concentration was positively associated with BALf epithelial cell percentage (P = 0.01). Air velocity >0.8 m/s was associated with increased odds of lung consolidation of ≥3 cm (OR = 6.8; CI = 1.2-38.5; P = 0.04) and ≥6 cm (OR = 15.9; CI = 1.2-200.0; P = 0.03). The prevalence of lung consolidations ≥1 cm was higher in the draught (81.8 %; P = 0.0092) and warm, dry and ammonia accumulation clusters (54.2 %; P = 0.02) compared to the presumably normal cluster (31.6 %). In addition, in the warm, dry and ammonia cluster the prevalence of lung consolidations ≥3 cm (38.1 %; P = 0.04) and ≥6 cm (31.4 %; P = 0.01) in depth were higher compared to the presumably normal climate cluster (18.2 % and 9.1 %, respectively). Of all frequently measured indoor air quality parameters, only average temperature, ammonia concentration and air velocity were associated with pneumonia and might therefore be preferable for cost-effective evaluation of calf barn climate.

Host Tolerance to Infection with the Bacteria that Cause Bovine Respiratory Disease.

Calves vary considerably in their pathologic and clinical responses to infection of the lung with bacteria. The reasons may include resistance to infection because of pre-existing immunity, development of effective immune responses, or infection with a minimally virulent bacterial strain. However, studies of natural disease and of experimental infections indicate that some calves develop only mild lung lesions and minimal clinical signs despite substantial numbers of pathogenic bacteria in the lung. This may represent "tolerance" to pulmonary infection because these calves are able to control their inflammatory responses or protect the lung from damage, without necessarily eliminating bacterial infection. Conversely, risk factors might predispose to bovine respiratory disease by triggering a loss of tolerance that results in a harmful inflammatory and tissue-damaging response to infection.

Bovine Respiratory Disease Diagnosis: What Progress Has Been Made in Infectious Diagnosis?

When it is desired to identify infectious agents involved in an outbreak of bovine respiratory disease, a variety of possible sampling methods may be used. For field use, the deep nasopharyngeal swab, transtracheal wash, and nonendoscopic bronchoalveolar lavage are most feasible. At present, bacterial culture and polymerase chain reaction testing are most commonly used to identify infectious agents. Interpretation of test results can be challenging, particularly for opportunistic pathogens. Evidence-based guidelines for precise interpretation of microbiologic tests results are lacking; however, approaches that have been practically useful for the management of bovine respiratory disease outbreaks are presented.

An evaluation of the economic effects of bovine respiratory disease on animal performance, carcass traits, and economic outcomes in feedlot cattle defined using four BRD diagnosis methods.

Bovine respiratory disease (BRD) causes significant economic losses to the feedlot industry due to decreased production and increased costs associated with treatment. This study aimed to assess the impacts of BRD on performance, carcass traits, and economic outcomes defined using four BRD diagnosis methods: number of BRD treatments an animal received, pleural lesions at slaughter, lung lesions at slaughter, and clinical BRD status defined using both treatment records and lung and pleural lesions. Crossbred steers (n = 898), with an initial body weight of 432 kg (± SD 51), were followed from feedlot entry to slaughter. Veterinary treatment records were collected and lungs scored at slaughter for lesions indicative of BRD. There was an 18% morbidity rate and a 2.1% BRD mortality rate, with an average net loss of AUD$1,647.53 per BRD mortality. Animals treated ≥3 times for BRD had 39.6 kg lighter carcasses at slaughter and returned an average of AUD$384.97 less compared to animals never treated for BRD (P < 0.001). Animals with severe lung lesions at slaughter grew 0.3 kg/d less, had 14.3 kg lighter carcasses at slaughter, and returned AUD$91.50 less than animals with no lung lesions (P < 0.001). Animals with subclinical and clinical BRD had 16.0 kg and 24.1 kg lighter carcasses, respectively, and returned AUD$67.10 and AUD$213.90 less at slaughter, respectively, compared to healthy animals that were never treated with no lesions (P < 0.001). The severity of BRD based on the number of treatments an animal received and the severity of lung and pleural lesions reduced animal performance, carcass weight and quality, and economic returns. Subclinical BRD reduced animal performance and economic returns compared to healthy animals; however, subclinical animals still had greater performance than animals with clinical BRD. This information can be used to plan for strategic investments aimed at reducing the impacts of BRD in feedlot cattle such as improved detection methods for subclinical animals with lesions at slaughter and BRD treatment protocols.

The pulmonary virome, bacteriological and histopathological findings in bovine respiratory disease from western Canada.

The aetiology and pathogenesis of bovine respiratory disease (BRD) are complex and involve the interplay of infectious agents, management and environmental factors. Previous studies of BRD focused on ante-mortem samples from the upper respiratory tract and identified several unconventional viruses. The lung, however, is the primary location where significant BRD lesions are usually found and is a common post-mortem diagnostic specimen. In this study, results of high-throughput virome sequencing, bacterial culture, targeted real-time PCR and histological examination of 130 bovine pneumonic lungs from western Canadian cattle were combined to explore associations of microorganisms with different types of pneumonia. Fibrinous bronchopneumonia (FBP) was the predominant type of pneumonia (46.2%, 60/130) and was associated with the detection of Mannheimia haemolytica. Detection of Histophilus somni and Pasteurella multocida was associated with suppurative bronchopneumonia (SBP) and concurrent bronchopneumonia and bronchointerstitial pneumonia (BP&BIP), respectively. Sixteen viruses were identified, of which bovine parvovirus 2 (BPV2) was the most prevalent (11.5%, 15/130) followed by ungulate tetraparvovirus 1 (UTPV1, 8.5%, 11/130) and bovine respiratory syncytial virus (BRSV, 8.5%, 11/130). None of these viruses, however, were significantly associated with a particular type of pneumonia. Unconventional viruses such as influenza D virus (IDV) and bovine rhinitis B virus (BRBV) were detected, although sparsely, consistent with our previous findings in upper respiratory tract samples. Taken together, our results show that while virus detection in post-mortem lung samples is of relatively little diagnostic value, the strong associations of H. somni and M. haemolytica with SBP and FBP, respectively, indicate that histopathology can be useful in differentiating bacterial aetiologies.

Serological investigation of antibodies against respiratory viruses in calves from Brazilian family farming and their relation to clinical signs of bovine respiratory disease / Investigação sorológica de anticorpos contra vírus respiratórios em bezerros de propriedades familiares brasileiras e sua relação com os sinais clínicos da doença respiratória dos bovinos

Bovine respiratory disease (BRD) is responsible for economic losses in cattle production. Viruses are categorized as primary etiological agents. The aims of this study were to evaluate the presence of antibodies against bovine viral diarrhea virus (BVDV), bovine herpes virus type 1 (BoHV-1), and bovine respiratory syncytial virus (BRSV) in healthy and BRD calves from family farming in relation to clinical signs of BRD. Hundred and forty-five calves were randomly selected and physical examination was performed. Only 123 animals were classified as healthy and BRD calves. Antibodies were evaluated by virus neutralization test. Person's Chi-square test and Fisher's exact test were performed as univariate analysis. Binary Logistic Regression was applied as multivariate analysis. Variables with P<0.10 were considered statistically significant. Variables with 0.15<P<0.10 were considered as statistical tendencies. Antibodies against BoHV-1, BVDV, and BRSV were detected in 32%, 23% and 37% animals. Antibodies against both three viruses were detected in 26.8% of calves. The presence of antibodies against BRSV were associated to normal heart rates (P=0.018) and unilateral airflow (P=0.035). Tendency was observed to unilateral airflow (P=0.06) Statistical tendencies were observed to Ab-BoHV-1 and body temperature (P=0.119) and breathing pattern (P=0.123). The profile of antibodies against respiratory viruses in calves from Brazilian family farming was firstly described. The results confirmed the importance of some clinical signs to viral infection.(AU)

A doença respiratória dos bovinos (DRB) é responsável por importantes perdas econômicas para a produção bovina, com maior impacto na agricultura familiar. Os vírus são comumente caracterizados como agentes etiológicos primários devido a mudanças na mucosa respiratória, na produção de citocinas e no funcionamento do sistema imune. Os objetivos deste estudo transversal foram avaliar a presença de anticorpos contra o vírus da diarreia viral bovina (VDVB), herpes vírus bovino tipo 1 (HVbo-1) e vírus respiratório sincicial bovino (VRSB) em bezerros sadios e com DRB de assentamentos em associação com a presença sinais clínicos de DRB. Cento e quarenta e cinco animais foram randomicamente selecionados e exame físico foi realizado. Somente 123 animais foram classificados em sadios e com DRB. Anticorpos foram identificados pelo teste de virusneutralização. Teste de qui-quadrado e teste exato de Fisher foram utilizados como análise univariada. A análise multivariada foi realizada pela regressão binária logística com o método Backward conditional. Variáveis com P<0,10 foram considerados significantes. Variáveis com 0,15<P<0,10 foram consideradas tendências. Anticorpos contra HVbo-1, VDVB e VRSB foram detectados em 32%, 23% e 37% dos animais. A presença concomitante de anticorpos contra os três vírus foi detectada em 26,8% dos bezerros. A presença de anticorpos contra VRSB foi associada à frequência cardíaca normal (P=0,018) e fluxo de ar nasal unilateral (P=0,035). Tendência estatística observada para anticorpos contra VDVB temperatura corporal (P=0,119) e padrão respiratório (P=0,123). Ao nosso conhecimento, este é o primeiro estudo que objetivou a avaliação da presença de anticorpos contra VDVB, VRSB e HVbo-1 em bezerros de assentamentos do estado de São Paulo, Brasil e sua relação com as manifestações clínicas da DRB.(AU)

Rapid Communication: Subclinical bovine respiratory disease - loci and pathogens associated with lung lesions in feedlot cattle.

Bovine respiratory disease (BRD) is an economically important disease of feedlot cattle that is caused by viral and bacterial pathogen members of the BRD complex. Many cases of subclinical BRD go untreated and are not detected until slaughter, when lung lesions are identified. The objectives of this study were to identify which BRD pathogens were associated with the presence of lung lesions at harvest and to identify genomic loci that were associated with susceptibility to lung lesions as defined by consolidation of the lung and/or the presence of fibrin tissue. Steers from a Colorado feedlot ( = 920) were tested for the presence of viral and bacterial pathogens using deep pharyngeal and mid-nasal swabs collected on entry into the study. Pathogen profiles were compared between cattle with or without lung consolidation (LC), fibrin tissue in the lung (FT), a combination of LC and FT in the same lung (lung lesions [LL]), and hyperinflated lungs (HIF) at harvest. Genotyping was conducted using the Illumina BovineHD BeadChip. Genomewide association analyses (GWAA) were conducted using EMMAX (efficient mixed-model association eXpedited), and pseudoheritabilities were estimated. The pathogen profile comparisons revealed that LC ( = 0.01, odds ratio [OR] = 3.37) and LL cattle ( = 0.04, OR = 4.58) were more likely to be infected with bovine herpes virus-1 and that HIF cattle were more likely to be infected with spp. ( = 0.04, OR = 4.33). Pseudoheritability estimates were 0.25 for LC, 0.00 for FT, 0.28 for LL, and 0.13 for HIF. Because pseudoheritability for FT was estimated to be 0, GWAA results for FT were not reported. There were 4 QTL that were moderately associated ( < 1 × 10) with only LC, 2 that were associated with only LL, and 1 that was associated with LC and LL. Loci associated with HIF included 12 that were moderately associated and 3 that were strongly associated (uncorrected P < 5 × 10-7). A 24-kb region surrounding significant lead SNP was investigated to identify positional candidate genes. Many positional candidate genes underlying or flanking the detected QTL have been associated with signal transduction, cell adhesion, or gap junctions, which have functional relevance to the maintenance of lung health. The identification of pathogens and QTL associated with the presence of lung abnormalities in cattle exhibiting subclinical BRD allows the identification of loci that may not be detected through manifestation of clinical disease alone.

An inactivated influenza D virus vaccine partially protects cattle from respiratory disease caused by homologous challenge.

Originally isolated from swine, the proposed influenza D virus has since been shown to be common in cattle. Inoculation of IDV to naïve calves resulted in mild respiratory disease histologically characterized by tracheitis. As several studies have associated the presence of IDV with acute bovine respiratory disease (BRD), we sought to investigate the efficacy of an inactivated IDV vaccine. Vaccinated calves seroconverted with hemagglutination inhibition titers 137-169 following two doses. Non-vaccinated calves challenged with a homologous virus exhibited signs of mild respiratory disease from days four to ten post challenge which was significantly different than negative controls at days five and nine post challenge. Peak viral shedding of approximately 5 TCID50/mL was measured in nasal and tracheal swabs and bronchoalveolar lavage fluids four to six days post challenge. Viral titers were significantly (P<0.05) decreased 1.4 TCID50/mL, 3.6 TCID50/mL and 5.0 TCID50/mL, respectively, in the aforementioned samples collected from vaccinated animals compared to non-vaccinated controls at peak shedding. Viral antigen was detected in the respiratory epithelium of the nasal turbinates and trachea by immunohistochemistry from all unvaccinated calves but in significantly fewer vaccinates. Inflammation characterized by neutrophils was observed in the nasal turbinate and trachea but not appreciably in lungs. Together these results support an etiologic role for IDV in BRD and demonstrate that partial protection is afforded by an inactivated vaccine.

Fever, feeding, and grooming behavior around peak clinical signs in bovine respiratory disease.

Feedlot cattle are monitored for the sickness response, both physiological and behavioral, to detect bovine respiratory disease (BRD), but this method can be inaccurate. Diagnostic accuracy may improve if the BRD sickness response is better understood. We hypothesized that steers around peak BRD would have fever, anorexia, and less grooming than controls. We also expected sickness response magnitude to be greater as clinical and pathological severity increased. Unvaccinated steers were assigned to challenge with 1 of 5 BRD viruses or bacteria (BRD challenge; = 4/pathogen; 20 total), based on susceptibility as determined by serology. Body weight-matched vaccinated animals were given sterile media (Control; = 4/pathogen; 20 total) and housed by treatment (5 pens/treatment). Rectal temperature was logged every 5 min between 0100 and 0700 h, and time spent feeding (24 h/d), in contact with a brush (13 h/d), and self-licking (24 h/d) were collected from video recordings. Steers were examined and a clinical score (CS) was assigned daily. Bovine respiratory disease challenge steers were euthanized after 5 to 15 d (timing was pathogen specific) and the proportion of grossly affected lung (%LUNG) was recorded. The day of highest CS (peak; d 0) for each BRD challenge steer and the 2 preceding days were analyzed for all variables except self-licking (d 0 only); analogous days were included for Controls. Penwise mixed models (pen was the experimental unit) were used to determine which sickness response elements differed between treatments before and at peak disease, and regression using individual-steer data was used to describe relationships between disease severity ( = 35 for CS and = 20 for %LUNG) and fever, anorexia, and grooming. Bovine respiratory disease challenge steers had fever (1.1°C higher; < 0.01) and anorexia (35% lower feeding time; = 0.03) but did not differ from healthy Controls for brush contact ( = 0.37) or self-licking ( = 0.15). Higher CS and more %LUNG were associated with increased fever (d 0; ≤ 0.04) and lower feeding (d 0; < 0.01), brush contact (d 0; ≤ 0.03), and self-licking ( ≤ 0.05) duration relative to lower CS and less %LUNG. In conclusion, fever and feeding time are good BRD diagnostic measures around peak CS. Further study is needed to clarify why grooming was not a good measure. The sickness response is greater as BRD severity increases; fever is most closely related to CS and anorexia is most closely related to %LUNG. Regardless of which aspect is monitored, cattle with milder disease may be more difficult to detect than sicker animals.

The upper respiratory tract microbiome and its potential role in bovine respiratory disease and otitis media.

The upper respiratory tract (URT) hosts a complex microbial community of commensal microorganisms and potential pathogens. Analyzing the composition and nature of the healthy URT microbiota and how it changes over time will contribute to a better understanding of the pathogenesis of pneumonia and otitis. A longitudinal study was conducted including 174 Holstein calves that were divided in four groups: healthy calves, calves diagnosed with pneumonia, otitis or both diseases. Deep pharyngeal swabs were collected on days 3, 14, 28, and 35 of life, and next-generation sequencing of the 16S rRNA gene as well as quantitative PCR was performed. The URT of Holstein dairy calves aged 3 to 35 days revealed to host a highly diverse bacterial community. The relative abundances of the bacterial genera Mannheimia, Moraxella, and Mycoplasma were significantly higher in diseased versus healthy animals, and the total bacterial load of newborn calves at day 3 was higher for animals that developed pneumonia than for healthy animals. Our results corroborate the existing knowledge that species of Mannheimia and Mycoplasma are important pathogens in pneumonia and otitis. Furthermore, they suggest that species of Moraxella can potentially cause the same disorders (pneumonia and otitis), and that high neonatal bacterial load is a key contributor to the development of pneumonia.

The association between calfhood bovine respiratory disease complex and subsequent departure from the herd, milk production, and reproduction in dairy cattle.

OBJECTIVE To describe the frequency of calfhood producer-identified bovine respiratory disease complex (BRDC) in Holstein replacement heifers on 1 large farm and determine associations between development of BRDC at ≤ 120 days of age (BRDC120) with milk production estimate, calving interval, and risk of departure from the herd (DFH). DESIGN Retrospective, observational study. ANIMALS 14,024 Holstein heifer calves born on 1 farm. PROCEDURES Data were obtained from herd management records. Cox proportional hazard and generalized linear mixed-effects models were used to assess associations for variables of interest (BRDC120 status, demographic data, and management factors) with DFH, milk production estimate, and calving interval. RESULTS Except for the year 2007, animals identified as having BRDC120 were 1.62 to 4.98 times as likely to leave the herd before first calving, compared with those that did not have this designation. Calves identified as having BRDC prior to weaning were 2.62 times as likely to have DFH before first calving as those classified as developing BRDC after weaning. Cows identified as having BRDC120 were 1.28 times as likely to have DFH between the first and second calving as were other cows. The BRDC120 designation was associated with a 233-kg (513-lb) lower 305-day mature equivalent value for first lactation milk production, but was not associated with longer or shorter calving intervals at maturity. CONCLUSIONS AND CLINICAL RELEVANCE Dairy cattle identified as having BRDC120 had increased risk of DFH before the first or second calving and lower first-lactation milk production estimates, compared with results for cattle without this finding. Further investigation of these associations is warranted.

Changes in feeding behavior patterns and dry matter intake before clinical symptoms associated with bovine respiratory disease in growing bulls.

Methods to improve accuracy of preclinical detection of bovine respiratory disease (BRD) are needed to reduce the economic impact of this disease, improve animal welfare, and promote more judicious use of antimicrobials in beef cattle. The objectives of this study were to retrospectively characterize time-series deviations in DMI and feeding behavior patterns preceding the display of observed clinical symptoms associated with BRD and to identify those feeding behavior traits that would be most predictive of BRD. The study was conducted with 231 seed stock bulls (391 ± 55 kg initial BW) representing 5 breeds that were housed in a facility equipped with GrowSafe feed bunks at a commercial bull-test facility. All bulls were vaccinated against standard viral and bacterial pathogens before and on arrival at the facility. Daily DMI and feeding behavior traits (frequency and duration of bunk visit events, head-down duration, variance of nonfeeding intervals, and time to approach feed bunk following feed-truck delivery) were measured for 70 d with a GrowSafe system. During a 10-d period from Day 28 to 37 of the trial, 30 bulls were administered antimicrobial therapy for clinical symptoms of BRD (rectal temperatures > 39.5°C). All remaining bulls ( = 201) were administered metaphylactic therapy on Day 38 of the trial in response to an acute decrease in feed intake. A retrospective analysis was conducted using a 2-slope broken-line regression model to identify inflection points in DMI and feeding behavior traits relative to onset of illness. The bulls were separated into 2 cohort groups based on observed clinical illness ( = 30) or those metaphylactically treated ( = 201), with the 2-slope broken-line regression model applied separately to each cohort. The model-detected inflection points for DMI were 6.8 and 3.8 d before observed clinical illness and metaphylactic treatment, respectively, and the reductions in DMI from detected inflection points to the day of observed clinical illness and day of metaphylactic treatment were 39.3 and 49.8%, respectively. Furthermore, the model-detected inflection points for individual feeding behavior traits ranged from 1.3 to 14.2 d before observed clinical illness and from 3.8 to 12.6 d before metaphylactic treatment. Results from this study demonstrate the potential value of electronic behavior-monitoring systems to improve the sensitivity and specificity of preclinical detection of BRD in feedlot cattle.

Treatment of Calves with Bovine Respiratory Disease: Duration of Therapy and Posttreatment Intervals.

When treating bovine respiratory disease, it is important to consider the decision to initiate treatment, the treatment regimen used, and whether to continue treatment. It is necessary to define the duration of drug exposure and when a success/failure decision will be made. No data are available to define the optimal duration of antimicrobial exposure. A pattern seen in human pneumonia studies is that shorter durations of therapy were equivalent with longer durations. Some studies suggest defining success or failure based on pharmacokinetics and pharmacodynamics may lead to earlier than optimal intervention. Optimal intervals are best defined by randomized clinical trials.

Genomewide association study of lung lesions in cattle using sample pooling.

Bovine respiratory disease complex (BRDC) is the most expensive disease in beef cattle in the United States costing the industry at least US$1 billion annually. Bovine respiratory disease complex causes damage to lung tissue resulting in persistent lung lesions observable at slaughter. Severe lung lesions at harvest have been associated with decreased preharvest ADG and increased clinical BRDC in the feedlot. Our objective was to identify SNP that are associated with severe lung lesions observed at harvest in feedlot cattle. We conducted a genomewide association study (GWAS) using a case-control design for severe lung lesions in fed cattle at slaughter using the Illumina Bovine HD array (approximately 770,000 SNP) and sample pooling. Lung samples were collected from 11,520 young cattle, a portion of which had not been treated with antibiotics (participating in a "natural" marketing program), at a large, commercial beef processing plant in central Nebraska. Lung samples with lesions (cases) and healthy lungs (controls) were collected when both phenotypes were in close proximity on the viscera (offal) table. We constructed 60 case and 60 control pools with 96 animals per pool. Pools were constructed by sampling sequence to ensure that case and control pool pairs were matched by proximity on the processing line. The Bovine HD array (770,000 SNP) was run on all pools. Fourteen SNP on BTA 2, 3, 4, 9, 11, 14, 15, 22, 24, and 25 were significant at the genomewide experiment-wise error rate of 5% ( ≤ 1.49 × 10). Eighty-five SNP on 28 chromosomes achieved a false discovery rate of 5% ( ≤ 5.38 × 10). Significant SNP were near (±100 kb) genes involved in tissue repair and regeneration, tumor suppression, cell proliferation, apoptosis, control of organ size, and immunity. Based on 85 significantly associated SNP in or near a collection of genes with diverse function on 28 chromosomes, we conclude that the genomic footprint of lung lesions is complex. A complex genomic footprint (genes and regulatory elements that affect the trait) is consistent with what is known about the cause of the disease: complex interactions among multiple viral and bacterial pathogens along with several environmental factors including dust, commingling, transportation, and stress. Characterization of sequence variation near significant SNP will enable accurate and cost effective genome-enhanced genetic evaluations for BRDC resistance in AI bulls and seed stock populations.