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.

Bovine Respiratory Disease Influences on Nutrition and Nutrient Metabolism.

Bovine respiratory disease (BRD) complex remains one of the greatest challenges facing beef cattle producers, veterinarians, and feedlot managers. In receiving, stocker/backgrounding, and feedlot cattle, BRD has been associated with decreased dry matter intake and daily gain, resulting in economic losses during the feeding period. Inflammation associated with BRD has the potential to decrease carcass yield and quality. Newly received calves are at various risks to contract BRD. Proper nutrition for newly received calves is key to recovery from stress associated with weaning and transport. This article reviews nutrient impacts on BRD and BRD impacts on nutrient metabolism.

Tissue Tropism in Host Transcriptional Response to Members of the Bovine Respiratory Disease Complex.

Bovine respiratory disease (BRD) is the most common infectious disease of beef and dairy cattle and is characterized by a complex infectious etiology that includes a variety of viral and bacterial pathogens. We examined the global changes in mRNA abundance in healthy lung and lung lesions and in the lymphoid tissues bronchial lymph node, retropharyngeal lymph node, nasopharyngeal lymph node and pharyngeal tonsil collected at the peak of clinical disease from beef cattle experimentally challenged with either bovine respiratory syncytial virus, infectious bovine rhinotracheitis, bovine viral diarrhea virus, Mannheimia haemolytica or Mycoplasma bovis. We identified signatures of tissue-specific transcriptional responses indicative of tropism in the coordination of host's immune tissue responses to infection by viral or bacterial infections. Furthermore, our study shows that this tissue tropism in host transcriptional response to BRD pathogens results in the activation of different networks of response genes. The differential crosstalk among genes expressed in lymphoid tissues was predicted to be orchestrated by specific immune genes that act as 'key players' within expression networks. The results of this study serve as a basis for the development of innovative therapeutic strategies and for the selection of cattle with enhanced resistance to BRD.

Pharmacokinetics and pharmacodynamics of gamithromycin in pulmonary epithelial lining fluid in naturally occurring bovine respiratory disease in multisource commingled feedlot cattle.

The objectives of this study were to determine (i) whether an association exists between individual pharmacokinetic parameters and treatment outcome when feeder cattle were diagnosed with bovine respiratory disease (BRD) and treated with gamithromycin (Zactran(®) ) at the label dose and (ii) whether there was a stronger association between treatment outcome and gamithromycin concentration in plasma or in the pulmonary epithelial lining fluid (PELF) effect compartment. The study design was a prospective, blinded, randomized clinical trial utilizing three groups of 60 (362-592 lb) steers/bulls randomly allocated within origin to sham injection or gamithromycin mass medication. Cattle were evaluated daily for signs of BRD by a veterinarian blinded to treatment. Animals meeting the BRD case definition were enrolled and allocated to a sample collection scheme consisting of samples for bacterial isolation (bronchoalveolar lavage fluid and nasopharyngeal swabs) and gamithromycin concentration determination (PELF and plasma). Gamithromycin susceptibility of M. haemolytica (n = 287) and P. multocida (n = 257) were determined using broth microdilution with frozen panels containing gamithromycin at concentrations from 0.03 to 16 µg/mL. A two-compartment plasma pharmacokinetic model with an additional compartment for gamithromycin in PELF was developed using rich data sets from published and unpublished studies. The sparse data from our study were then fit to this model using nonlinear mixed effects modeling to estimate individual parameter values. The resulting parameter estimates were used to simulate full time-concentration profiles for each animal in this study. These profiles were analyzed using noncompartmental methods so that PK/PD indices (AUC24 /MIC, AUC∞ /MIC, CMAX /MIC) could be calculated for plasma and PELF (also T>MIC) for each individual. The calculated PK/PD indices were indicative that for both M. haemolytica and P. multocida a higher drug exposure in terms of concentration, and duration of exposure relative to the MIC of the target pathogen, was favorable to a successful case outcome. A significant association was found between treatment success and PELF AUC0-24 /MIC for P. multocida. The calves in this study demonstrated an increased clearance and volume of distribution in plasma as compared to the healthy calves in two previously published reports. Ultimately, the findings from this study indicate that higher PK/PD indices were predictive of positive treatment outcomes.

Acute phase proteins in naturally occurring respiratory disease of feedlot cattle.

The aim of this study was to evaluate three acute phase proteins (APP) [haptoglobin (HPT), lipopolysaccharide binding protein (LBP) and transferrin (Tf)] in feedlot cattle with naturally occurring respiratory disease diagnosed by a calf health scoring chart (CHSC). Seventy-seven beef calves were observed for signs of Bovine Respiratory Disease (BRD) during the first 28 days after arrival at the feedlot. Fourteen cases and pen matched controls were selected based on the CHSC. BRD cases were defined as a score of ≥ 5, while controls were defined as a score ≤ 4. The mean CHSC score in cases was 6.9 which was significantly greater than the controls 2.8 (P < 0.01). Mean plasma LBP and HPT concentrations were significantly greater in cases than controls (P < 0.01). Our study results show that measurement of HPT and LBP could be useful in detecting respiratory disease in feedlot conditions. Transferrin concentrations between the two groups were not statistically different.

Differential stress responses among newly received calves: variations in reductant capacity and Hsp gene expression.

Bovine respiratory disease complex (BRD), a major economic concern to the beef cattle industry all over the world, is triggered by physical, biological and psychological stresses. It is becoming noticeable that the key to reducing BRD appears to be centered at reducing the response to stress. The aims of the present study were to detect individual variations in the stress response of newly received young calves through their leukocyte heat shock protein (Hsp) response, selected neutrophil-related gene expression and oxidative stress, and relate them to pulmonary adhesions at slaughter, an indicative sign of clinical and subclinical episodes of BRD at an early age. Differential expression patterns of Hsp60 and Hsp70A1A were revealed in newly received calves 1 h, 5 h and 1 day after arrival, distinguishing between stress-responsive and non-stress-responsive individuals. Plasma cortisol was also indicative of stress-responsive and non-stress-responsive individuals, 1 h and 5 h after arrival. At the longer term, ß-glycan levels were highest 7 days after arrival and significantly correlated with an adhesion-free phenotype at slaughter. Oxidative stress responses, measured through the oxidation products of the exogenous linoleoyl tyrosine (LT) marker, revealed that hydroperoxidation and epoxidation of membranes may readily occur. Based on the LT oxidation products and levels of ß-glycan, we present a discriminant analysis model, according to which vulnerable individuals may be predicted at near 100% probability 7 days after arrival. Since clinical signs of BRD may often go undetected in feedlot calves, such a model, after its examination in large-scale experiments, may be a reliable tool for an early prediction of subclinical signs of BRD.

Effect of bovine respiratory disease during preconditioning on subsequent feedlot performance, carcass characteristics, and beef attributes.

Heifers with expected increased risk of bovine respiratory disease (BRD; n = 360; initial BW = 241.3 +/- 16.6 kg) were assembled at a Kentucky order-buyer facility and delivered to Stillwater, OK, in September 2007 to determine the effects of clinical BRD observed during preconditioning on subsequent feedlot performance, carcass characteristics, and meat attributes. During a 63-d preconditioning period, morbidity and mortality attributed to BRD were 57.6 and 8.6%, respectively. Immediately after preconditioning, heifers were grouped according to health outcome category and allotted to finishing pens (5 to 7 heifers/pen). Heifers were never treated for BRD (0X; n = 9 pens), treated 1 time (1X; n = 9 pens), 2 times (2X; n = 6 pens), 3 times (3X; n = 6 pens), or designated as chronically ill (CI; n = 2 pens). Arrival BW was not different (P = 0.21) among treatment categories. However, disease incidence during preconditioning decreased (P < 0.001) growth, resulting in BW of 318, 305, 294, 273, and 243 kg for 0X, 1X, 2X, 3X, and CI, respectively, at the start of the finishing phase. Estimates on the LM, taken by ultrasound on d 65 and 122, were combined with BW and visual appraisal to target common average endpoint within category and block. On average, heifers were slaughtered on d 163 for 0X, 1X, and 2X, d 182 for 3X, and d 189 for CI (P < 0.01). Final BW was similar (P > or = 0.18) for heifers treated 0, 1, 2, or 3 times, but heifers deemed CI weighed less (P = 0.01) than 3X heifers. Considering the finishing phase only, ADG was linearly increased (P < 0.001) with increasing BRD treatments, but was linearly decreased (P = 0.003) as BRD treatments increased from arrival to slaughter. Therefore, G:F was greater (P = 0.007) for CI than 3X and linearly increased (P = 0.002) from 0X to 3X. Similar to BW, HCW was less (P = 0.03) for CI than 3X. Marbling score tended (P = 0.06) to decrease linearly as the number of treatments increased, but no other differences (P > or = 0.24) in carcass traits were detected. No differences were observed in beef tenderness (P = 0.65), and no consistent trends were noted in retail display or palatability data. Less than 20 additional days on feed were required for heifers treated 3 times to have similar BW and carcass characteristics to heifers never treated for BRD. Segregating animals with multiple BRD treatments and feeding them to an acceptable carcass endpoint may be a viable strategy for increasing value of animals treated for BRD.

Biomarkers for prediction of bovine respiratory disease outcome.

Fatal bovine respiratory disease (BRD) is frequently the result of a primary viral and a secondary bacterial respiratory infection. In cattle, BRD causes more than half of feedlot deaths and has a major impact on financial losses in the cattle industry in North America. It is, therefore, very important to understand the mechanism of this complex disease process as well to predict and identify BRD susceptible cattle to enhance treatment efficacy. We recently established the value of using combinatorial omics approaches to identify candidate biomarkers associated with stress responses, a factor that can increase the severity of BRD. The objective of the present investigation was to experimentally recreate fatal BRD and to use a combinatorial analysis of proteomic, metabonomic, and elemental profiles in serum samples to determine if multimethod analysis of these biomarkers could predict disease outcome. The proteomic studies revealed that changes in the serum proteome were significant on day 4 postviral infection when compared to preinfection (day 0) serum samples. Proteomic studies identified a group of acute phase proteins (haptoglobin and apolipoprotein AI), which could be linked to a primary viral respiratory infection, but there was no significant association observed with fatal BRD. In contrast, metabonomic and elemental analyses identified candidate biomarkers for viral infection (glucose, LDL, valine, phosphorous, and iron) and disease outcome (lactate, glucose, iron). While multivariate analysis of proteomic and metabolite profiles did not discriminate between animals that survived or died postsynergic viral-bacterial infection by analyzing preinfection (day 0) serum samples, analysis of serum elemental profiles prior to infection was, however, predictive of BRD outcome. Furthermore, discriminant analyses of all three methodologies used to profile serum (collected on day 4 postviral but prior to bacterial infection) revealed differential trends between animals that survived or died following synergic viral-bacterial infection. Thus, a combinatorial approach using proteomic, metabonomic, and elemental analyses of serum samples revealed that multimethod analysis could discriminate between the complex biological responses to secondary bacterial respiratory infection and predict disease outcome.