Early detection of bovine respiratory disease in pre-weaned dairy calves using sensor based feeding, movement, and social behavioural data.

Previous research shows that feeding and activity behaviours in combination with machine learning algorithms has the potential to predict the onset of bovine respiratory disease (BRD). This study used 229 novel and previously researched feeding, movement, and social behavioural features with machine learning classification algorithms to predict BRD events in pre-weaned calves. Data for 172 group housed calves were collected using automatic milk feeding machines and ultrawideband location sensors. Health assessments were carried out twice weekly using a modified Wisconsin scoring system and calves were classified as sick if they had a Wisconsin score of five or above and/or a rectal temperature of 39.5 °C or higher. A gradient boosting machine classification algorithm produced moderate to high performance: accuracy (0.773), precision (0.776), sensitivity (0.625), specificity (0.872), and F1-score (0.689). The most important 30 features were 40% feeding, 50% movement, and 10% social behavioural features. Movement behaviours, specifically the distance walked per day, were most important for model prediction, whereas feeding and social features aided in the model's prediction minimally. These results highlighting the predictive potential in this area but the need for further improvement before behavioural changes can be used to reliably predict the onset of BRD in pre-weaned calves.

Use of fidget and drinking behaviour in combination with facial infrared thermography for diagnosis of bovine respiratory disease in a spontaneous model.

Bovine respiratory disease (BRD) is a highly prevalent multi pathogen infectious disease (70-80%) in newly received feedlot cattle, causing significant economic losses and reduced animal welfare. Current BRD diagnosis involves stressful and invasive methods that can increase the incidence and transmission of BRD. An alternative is the use of an automated infrared thermography (IR) platform that can monitor facial temperature and behaviour traits to diagnose BRD in a non-invasive manner. The objective of this study was to investigate the use of fidget and drinking behaviours in conjunction with facial temperature as method of BRD diagnosis in beef calves. Sixty-five weaned calves (N = 65) were monitored over a 21-d period after 6 h transportation to predispose calves to BRD infection. Data collected from an automated IR platform placed at a water station included the number of IR frames during drinking (Fidget), number of drinking visits (Drinking bouts), total drinking duration, average drinking duration, average cheek temperature (AVG temp), and maximum orbital temperature (Max temp). Fidget, drinking behaviours, and IR were compared to a clinical score assessment based on respiratory, digestive, and lethargy signs (visual observation) and haematology analysis using a receiver operating characteristics curve analysis to identify the accuracy of each metric and combinations of metrics for BRD diagnosis. The greater accuracies observed were Fidget, Youden's index (J): 0.25 J), Drinking bout (0.28 J), and Total drinking duration (0.22 J). The average IR temperature accuracy resulted in 0.88 J and Max temp 0.77 J. Thirty-five combinations of drinking behaviour and facial IR metrics were evaluated to identify BRD calves. Optimum accuracy (100%) was achieved when combining Fidget, Drinking bout, Average drinking duration, AVG temp, and Max temp 1.00 J. Similar evaluations were performed at 48 and 24 h before d 0 using the most accurate Fidget, Drinking behaviour, and IR combination, resulting in 0.44 J 48 h prior to d 0 and 0.45 J 24 h prior to d 0. Combining fidget and drinking behaviour metrics increased the sensitivity to detect the onset of BRD infection and the specificity to discriminate true positive BRD calves from true negative BRD calves.

An overview of the detection of bovine respiratory disease complex pathogens using immunohistochemistry: emerging trends and opportunities.

The bovine respiratory disease complex (BRDC) is caused by a variety of pathogens, as well as contributing environmental and host-related risk factors. BRDC is the costliest disease for feedlot cattle globally. Immunohistochemistry (IHC) is a valuable tool for enhancing our understanding of BRDC given its specificity, sensitivity, cost-effectiveness, and capacity to provide information on antigen localization and immune response. Emerging trends in IHC include the use of multiplex IHC for the detection of coinfections, the use of digital imaging and automation, improved detection systems using enhanced fluorescent dyes, and the integration of IHC with spatial transcriptomics. Overall, identifying biomarkers for early detection, utilizing high-throughput IHC for large-scale studies, developing standardized protocols and reagents, and integrating IHC with other technologies are some of the opportunities to enhance the accuracy and applicability of IHC. We summarize here the various techniques and protocols used in IHC and highlight their current and potential role in BRDC research.

Validation of a multiplex-tandem RT-PCR for the detection of bovine respiratory disease complex using Scottish bovine lung samples.

The welfare and economic impact of bovine respiratory disease complex (BRDC), and its associated antibiotic usage, are major challenges to cattle rearing and beef cattle finishing industries. Accurate pathogen diagnosis is important to undertake appropriate treatment and long-term management strategies, such as vaccine selection. Conventional diagnostic approaches have several limitations including high costs, long turnaround times and difficulty in test interpretation, which could delay treatment decisions and lead to unnecessary animal losses. We describe the validation of a multiplex-tandem (MT) reverse transcription-polymerase chain reaction (RT-PCR) for the detection of seven common pathogens associated with BRDC. This test has the potential to advance pathogen identification and to overcome many of the limitations of current testing methods. It requires a single sample and results are obtained quickly and not influenced by prior antimicrobial therapy or overgrowth of contaminating organisms. We demonstrated a test specificity of 100% and sensitivity ranging from 93.5% to 100% for these seven common pathogens. This test will be a useful addition to advance BRDC investigation and diagnosis.

Modelling the effects of antimicrobial metaphylaxis and pen size on bovine respiratory disease in high and low risk fattening cattle.

Bovine respiratory disease (BRD) dramatically affects young calves, especially in fattening facilities, and is difficult to understand, anticipate and control due to the multiplicity of factors involved in the onset and impact of this disease. In this study we aimed to compare the impact of farming practices on BRD severity and on antimicrobial usage. We designed a stochastic individual-based mechanistic BRD model which incorporates not only the infectious process, but also clinical signs, detection methods and treatment protocols. We investigated twelve contrasted scenarios which reflect farming practices in various fattening systems, based on pen sizes, risk level, and individual treatment vs. collective treatment (metaphylaxis) before or during fattening. We calibrated model parameters from existing observation data or literature and compared scenario outputs regarding disease dynamics, severity and mortality. The comparison of the trade-off between cumulative BRD duration and number of antimicrobial doses highlighted the added value of risk reduction at pen formation even in small pens, and acknowledges the interest of collective treatments for high-risk pens, with a better efficacy of treatments triggered during fattening based on the number of detected cases.

Diagnostic accuracy of Wisconsin and California scoring systems to detect bovine respiratory disease in preweaning dairy calves under subtropical environmental conditions.

Bovine respiratory disease (BRD) is a multifactorial disease which causes short- and long-term negative effects. Early detection is crucial for a prompt response to therapy, as well as to decrease mortality risk. Clinical scoring systems have been developed mostly in North America for screening calves at risk or suspected of having BRD, and these tools have also been applied in subtropical and tropical countries. However, it has been unknown whether these scoring systems had the same accuracy in tropical environmental conditions. Therefore, this study evaluated the accuracy of 4 different field techniques, as well as serum haptoglobin (HAP), to diagnose BRD in Holstein dairy calves in subtropical conditions. The tests used to diagnose BRD were thoracic ultrasound (TUS; positive if consolidation depth ≥1 cm), thoracic auscultation (AUSC; positive if crackles, wheezes, or silent areas were present), Wisconsin score (WISC; ≥2 categories with scores of ≥2), and California score (CALIF; positive if total score ≥5). Also, HAP was measured and classified as positive if ≥15 mg/dL. Heifers between 30 d of age and weaning (n = 482), residing on 17 commercial dairies in São Paulo state, were enrolled in this study. Bayesian latent class models were used with informative priors to evaluate the accuracy of TUS, AUSC, and HAP, and noninformative priors for the accuracy of WISC and CALIF. The percentage of calves positive for each test on each farm ranged from 0 to 56% for WISC, 11-51% for CALIF, 0-72% for TUS, 0-32% for AUSC, and 0-100% for HAP. The sensitivity (Se; 95% credible interval) and specificity (Sp) for WISC were 77.9% (64.8-90.2) and 81.9% (76.3-88.2). For CALIF, the Se was 67.1% (53.6-80.1) and Sp 79.1% (73.9-84.6). For TUS Se was 59.8% (46.5-73.1) and Sp was 84.8% (80.0-89.5), and for AUSC, Se was 58.8% (41.3-79.8) and Sp was 98.6% (95.7-99.9). The Se and Sp of HAP was 67.6% (55.3-78.8) and 46.7% (41.4-52.2), respectively. The performance of the scoring systems was similar to, or better than, the performance found in North American studies, despite the fact that calves were in a tropical environment.

Daily behavioral measures recorded by precision technology devices may indicate bovine respiratory disease status in preweaned dairy calves.

Precision technology devices can measure and detect relative changes in an animal's behavior to possibly create alerts to intervene and to administer treatments. However, the association of relative changes in daily feeding and activity behaviors in calves with bovine respiratory disease (BRD) status is still largely unexplored. The objective of this case-control study was to determine if daily behavioral patterns of preweaning dairy calves (measured by precision technologies) change before BRD diagnosis. This case-control study enrolled 33 pairs of calves (33 BRD calves matched by age, sex, and birthdate to 33 controls) health scored daily for BRD for the preweaning period (until 50 d on the automated feeder). A pedometer (IceQube, Ice Robotics) was attached to the left rear leg to track activity (lying time, lying bouts, total steps, and acceleration activity index). At 3.0 ± 2.0 d of age, calves were trained to use the automated feeder, which recorded milk and calf starter intake, drinking speed, and feeder visits. Calves were allotted a maximum 10 L/d of milk replacer (Cow's Match, Land O'Lakes Animal Milk Products Co.; 140 g/L) and starter (Special Calf Starter and Grower, Baghdad Feeds), ad libitum. Calves were scored daily for signs of BRD using the Wisconsin scoring system, and their lungs were scored with ultrasonography twice weekly. Outward signs of BRD as defined by the Wisconsin scoring system and an area of consolidated lung ≥3.0 cm2 was identified as BRD (d 0). Relative changes in daily behaviors were calculated using d -5 before BRD diagnosis as the baseline for each calf for each behavior. Linear mixed models were used to investigate the association of BRD status with feeding behaviors, activity, and relative changes in calf behavior over a 5-d period before diagnosis, as well as the BRD status × day interaction. Calves with a BRD bout were diagnosed at an average age of 33.0 ± 9.0 d (mean ± standard deviation) and weighed 56.1 ± 9.7 kg. Over the period, BRD calves had reduced milk and starter intake, greater lying times, and fewer lying bouts, step counts, and activity indices when compared with healthy calves. Furthermore, there was a BRD status × day interaction for relative changes in unrewarded visits, and relative changes in calf starter intake. Specifically, BRD calves had a decline in relative changes in their unrewarded visits on d -4, -2, -1, and 0 compared with healthy calves. This study suggests that there is the potential to use feeding and activity behaviors to identify BRD development in preweaning calves. However, utilization of an animals' daily behavioral patterns in real time is fundamental for developing disease detection algorithms, thus we suggest relative changes in unrewarded visits may be useful for algorithm development when d -5 is used as a baseline. Future research should investigate the potential of feeding behavior and activity levels collectively to indicate BRD status in calves using machine learning techniques.

A systematic review of the utility of biomarkers as aids in the early diagnosis and outcome prediction of bovine respiratory disease complex in feedlot cattle.

Bovine respiratory disease complex (BRDC) is a common, serious problem in feedlot cattle worldwide. Early diagnosis and outcome prediction are critical for making decisions to prevent economic loss and to limit antimicrobial use. Diagnosing BRDC is commonly based on visual signs and behavioral changes; both assessments are considered to have low diagnostic accuracy. Biomarkers are important for supporting the diagnosis of BRDC, determining the necessity and potential outcomes of treatment, and assisting in research in which differentiating diseased animals is required. There are few reviews summarizing the biomarkers available and utilized. We systematically evaluated the detection and prognostic potential of biomarkers from the literature published between January 1990 and December 2020. We performed a descriptive analysis of 5 biomarker categories: acute-phase proteins, stress-related hormones, other blood biomarkers, omics biomarkers, and non-blood biomarkers. The retrieved articles consisted of studies or trials that assessed the detection value and treatment and/or outcome prediction efficacy of biomarkers for BRDC in feedlot cattle; 23 manuscripts for review and analysis satisfied the selection criteria. Based on our review, we cannot recommend a specific biomarker as the sole method for the early detection or outcome prediction for BRDC, given that the application and efficacy of biomarkers varies in different situations. Our systematic review may serve as a reference for clinical and research investigations of early detection and outcome prediction of BRDC.

Galyean Appreciation Club Review: revisiting nutrition and health of newly received cattle-what have we learned in the last 15 years?

Our objective was to review the literature related to the health and management of newly received cattle published since a previous review by Duff and Galyean (2007). Bovine respiratory disease (BRD) continues to be a major challenge for the beef industry. Depending on disease severity, animals treated for BRD have decreased performance and lowered carcass value. Diagnosis of BRD is less effective than desired, and progress on developing real-time, chute-side methods to diagnose BRD has been limited. Systems that combine lung auscultation with temperature and body weight data show promise. Assessment of blood metabolites and behavior monitoring offer potential for early identification of morbid animals. Vaccination and metaphylaxis continue to be important tools for the prevention and control of BRD, but antimicrobial resistance is a concern with antibiotic use. Dietary energy concentration and roughage source and level continue to be important topics. Mineral supplementation has received considerable attention, particularly the use of organic vs. inorganic sources and injectable minerals or drenches given on arrival. The use of probiotics and prebiotics for newly received cattle has shown variable results, but further research is warranted. The health and nutrition of newly received cattle will continue to be an important research area in the years to come.

Bovine respiratory disease (BRD) is a significant economic and animal welfare challenge for the beef industry. Experiments related to the health and management of newly received cattle published in the last 15 yr were reviewed. Limited progress is being made in developing accurate, real-time methods for diagnosis of BRD, and overall, diagnosis is less effective than desired. Measurement of lung and heart sounds combined with rectal temperature have been studied as diagnostic tools, as well as measurement of blood metabolites and remote monitoring of behavior. Vaccination for viral and bacterial BRD agents and mass treatment of cattle with antibiotics continue to be important tools for prevention and control of BRD, but the development of antimicrobial resistance is a concern. Energy and roughage concentration as well as roughage source continue to be important dietary considerations, as does mineral supplementation, with mineral source and injectable minerals receiving significant research attention. Probiotics and prebiotics fed to newly received cattle have shown variable results in terms of effects on the incidence of BRD and animal performance. Additional research is needed to define optimal diagnostic, management, and nutritional practices for newly received cattle.

Use of nCounter mRNA profiling to identify at-arrival gene expression patterns for predicting bovine respiratory disease in beef cattle.

BACKGROUND: Transcriptomics has identified at-arrival differentially expressed genes associated with bovine respiratory disease (BRD) development; however, their use as prediction molecules necessitates further evaluation. Therefore, we aimed to selectively analyze and corroborate at-arrival mRNA expression from multiple independent populations of beef cattle. In a nested case-control study, we evaluated the expression of 56 mRNA molecules from at-arrival blood samples of 234 cattle across seven populations via NanoString nCounter gene expression profiling. Analysis of mRNA was performed with nSolver Advanced Analysis software (p < 0.05), comparing cattle groups based on the diagnosis of clinical BRD within 28 days of facility arrival (n = 115 Healthy; n = 119 BRD); BRD was further stratified for severity based on frequency of treatment and/or mortality (Treated_1, n = 89; Treated_2+, n = 30). Gene expression homogeneity of variance, receiver operator characteristic (ROC) curve, and decision tree analyses were performed between severity cohorts. RESULTS: Increased expression of mRNAs involved in specialized pro-resolving mediator synthesis (ALOX15, HPGD), leukocyte differentiation (LOC100297044, GCSAML, KLF17), and antimicrobial peptide production (CATHL3, GZMB, LTF) were identified in Healthy cattle. BRD cattle possessed increased expression of CFB, and mRNA related to granulocytic processes (DSG1, LRG1, MCF2L) and type-I interferon activity (HERC6, IFI6, ISG15, MX1). Healthy and Treated_1 cattle were similar in terms of gene expression, while Treated_2+ cattle were the most distinct. ROC cutoffs were used to generate an at-arrival treatment decision tree, which classified 90% of Treated_2+ individuals. CONCLUSIONS: Increased expression of complement factor B, pro-inflammatory, and type I interferon-associated mRNA hallmark the at-arrival expression patterns of cattle that develop severe clinical BRD. Here, we corroborate at-arrival mRNA markers identified in previous transcriptome studies and generate a prediction model to be evaluated in future studies. Further research is necessary to evaluate these expression patterns in a prospective manner.

Bovine Respiratory Disease Considerations in Young Dairy Calves.

Raising young dairy calves presents many challenges for producers and veterinarians including losses attributable to BRD. This article will discuss several key concepts for practitioners to consider when applying evidence-based medicine for the control and treatment of BRD in young dairy calves. The authors review BRD complex, provide considerations for diagnostic approaches, and discuss research associated with the control and treatment of BRD.

Assessment of diagnostic accuracy of biomarkers to assess lung consolidation in calves with induced bacterial pneumonia using receiver operating characteristic curves.

Bovine respiratory disease (BRD) is the most economically significant disease for cattle producers in the U.S. Cattle with advanced lung lesions at harvest have reduced average daily gain, yield grades, and carcass quality outcomes. The identification of biomarkers and clinical signs that accurately predict lung lesions could benefit livestock producers in determining a BRD prognosis. Receiver operating characteristic (ROC) curves are graphical plots that illustrate the diagnostic ability of a biomarker or clinical sign. Previously we used the area under the ROC curve (AUC) to identify cortisol, hair cortisol, and infrared thermography imaging as having acceptable (AUC > 0.7) diagnostic accuracy for detecting pain in cattle. Herein, we used ROC curves to assess the sensitivity and specificity of biomarkers and clinical signs associated with lung lesions after experimentally induced BRD. We hypothesized pain biomarkers and clinical signs assessed at specific time points after induction of BRD could be used to predict lung consolidation at necropsy. Lung consolidation of > 10% was retrospectively assigned at necropsy as a true positive indicator of BRD. Calves with a score of < 10% were considered negative for BRD. The biomarkers and clinical signs analyzed were serum cortisol; infrared thermography (IRT); mechanical nociceptive threshold (MNT); substance P; kinematic gait analysis; a visual analog scale (VAS); clinical illness score (CIS); computerized lung score (CLS); average activity levels; prostaglandin E2 metabolite (PGEM); serum amyloid A; and rectal temperature. A total of 5,122 biomarkers and clinical signs were collected from 26 calves, of which 18 were inoculated with M. haemolytica. All statistics were performed using JMP Pro 14.0. Results comparing calves with significant lung lesions to those without yielded the best diagnostic accuracy (AUC > 0.75) for right front stride length at 0 h; gait velocity at 32 h; VAS, CIS, average activity and rumination levels, step count, and rectal temperature, all at 48 h; PGEM at 72 h; gait distance at 120 h; cortisol at 168 h; and IRT, right front force and serum amyloid A, all at 192 h. These results show ROC analysis can be a useful indicator of the predictive value of pain biomarkers and clinical signs in cattle with induced bacterial pneumonia. AUC values for VAS score, average activity levels, step count, and rectal temperature seemed to yield good diagnostic accuracy (AUC > 0.75) at multiple time points, while MNT values, substance P concentrations, and CLS did not (all AUC values < 0.75).

Bovine respiratory disease (BRD) is the most economically significant disease for cattle producers in the United States, affecting 16.2% of cattle on feed. Cattle with advanced lung lesions at harvest have reduced average daily gain, yield grades, and carcass quality outcomes. The identification of biomarkers and clinical signs that accurately predict lung lesions could benefit livestock producers in determining a BRD prognosis. Herein, we used receiver operating characteristic curves to assess the predictive value of biomarkers and clinical signs associated with lung lesions after experimentally induced BRD. In the first 72 h after onset of BRD, right front stride length, gait velocity, visual analog scale score, clinical illness score, average activity level, step count, and rectal temperature yielded the best diagnostic accuracy (AUC > 0.75) for predicting calves with significant lung lesions (>10% consolidation) at necropsy. Biomarkers and clinical signs with the best diagnostic accuracy early in the disease process would likely be the most valuable in field conditions. These results can be used to guide refinement of the optimal time points and biomarkers for the diagnosis of significant lung lesions after BRD.

Evaluation of reticulorumen temperature boluses for the diagnosis of subclinical cases of bovine respiratory disease in feedlot cattle.

Bovine respiratory disease (BRD) is the most important and costly health issue of the feedlot industry worldwide. Remote monitoring of reticulorumen temperature has been suggested as a potential tool to improve the diagnostic accuracy of BRD. The present study aimed to evaluate 1) the difference and degree of reticulorumen hyperthermia episodes between healthy and subclinical BRD feedlot steers, and 2) determine the correlation between reticulorumen hyperthermia and lung pathology, performance, and carcass traits. Mixed-breed feedlot steers (n = 148) with a mean arrival weight of 321 ± 3.34 kg were administered a reticulorumen bolus at feedlot entry and monitored for visual and audible signs of BRD until slaughter when lungs were examined and scored for lesions indicative of BRD. Post-slaughter animals with no record of BRD treatment were assigned to one of three case definitions. Healthy steers had no visual or audible signs of BRD (i.e., CIS=1), and total lung consolidation score < 5% or pleurisy score < 3 at slaughter. Subclinical BRD cases had a CIS of 1, and a lung consolidation score ≥ 5% or a pleurisy score of 3 at slaughter. Mild CIS cases had at least one CIS of 2, and a lung consolidation score < 5% and a pleurisy score < 3 at slaughter. Subclinical BRD and mild CIS cases had longer total duration of reticulorumen hyperthermia, more episodes and longer average episode duration above 40.0 °C compared to healthy steers (P < 0.05). A moderate positive correlation was found between lung consolidation and total duration (r = 0.27, P < 0.001), episode duration (r = 0.29, P < 0.001), and number of episodes (r = 0.20, P < 0.05). Pleurisy score was also found to be moderately and positively correlated with total duration (r = 0.23, P < 0.01), episode duration (r = 0.37, P < 0.001), and number of episodes (r = 0.26, P < 0.01). Moderate negative correlations were found between reticulorumen hyperthermia and carcass traits including hot standard carcass weight (HSCW) (-0.22 ≤ r ≤ -0.23, P < 0.05) and P8-fat depth (-0.18 ≤ r ≤ -0.32, P < 0.05). Subclinical BRD reduced carcass weight by 22 kg and average daily gain (ADG) by 0.44 kg/day compared to healthy steers (P < 0.05), but mild CIS cases had no effect on performance (P > 0.05). The reticulorumen bolus technology appears promising for detection of subclinical BRD cases in feedlot cattle as defined by lung pathology at slaughter.

Scoping review on clinical definition of bovine respiratory disease complex and related clinical signs in dairy cows.

Bovine respiratory disease complex (BRD) is a worldwide multifactorial infectious disease. Antimicrobials are commonly used for treating BRD because bacteria are often involved. The clinical diagnosis of BRD is a challenge, especially in adult dairy cows, where information on this syndrome is scant. Having a definition based on consistent and reliable clinical signs would improve the accuracy of BRD diagnosis and could help to develop an optimal treatment approach by an early detection. The aim of this scoping review was to review clinical signs that could be recognized by producers in dairy cattle suffering from naturally occurring infectious respiratory disease, as reported in the literature. A review of the literature was performed for articles published between January 1, 1990 and January 1, 2020. The search of literature in English, French, and Italian languages included 2 different databases (Pubmed, https://pubmed.ncbi.nlm.nih.gov/; CAB abstract, https://www.cabi.org/publishing-products/cab-abstracts/). Clinical signs were categorized as follows: (1) "general manifestations of disease," which included behavioral changes or fever; (2) "alterations in respiratory function," which included clinical signs specifically associated with the respiratory tract examination; and (3) "clinical signs of other body systems," which included clinical signs related to other systems such as diarrhea or subcutaneous emphysema. The focus of the review was on clinical signs that could be monitored by animal handlers and producers. A total of 1,067 titles were screened, and 23 studies were finally included. The most common general clinical signs were increased body temperature (reported in 83% of studies, n = 19), change in feed intake (26%, n = 6), altered mentation (22%, n = 5), and decreased milk production (17%, n = 4). The alterations in respiratory function noted were nasal discharge (74%, n = 17), cough (65%, n = 15), altered respiratory dynamic or dyspnea (61%, n = 14), increased respiratory rate (43%, n = 10), and ocular discharge or lacrimation (30%, n = 7). The clinical signs associated with infectious respiratory disease reported in the 23 studies generally lacked a clear description of what constitutes a deviation from normality (0-50% of studies clearly reported what was considered normal versus abnormal depending on the clinical signs). This limitation prevented any comparison between studies that apparently reported the same "clinical sign," but possibly referred to a different assessment and definition of what was considered normal versus abnormal. Therefore, the definition of clinical signs in a repeatable way with validated interobserver agreement to determine the optimal combination for the diagnosis of BRD in dairy cows is needed. This could lead to a more judicious use of antimicrobials for respiratory disease in adult dairy cows.

Molecular characterization and comparison of diagnostic methods for bovine respiratory viruses (BPIV-3, BRSV, BVDV, and BoHV-1) in field samples in northwestern Turkey.

The aim of this study was to evaluate the compatibility among virus isolation (VI), ELISA, and PCR for diagnosis of the major viral agents (BPIV-3, BRSV, BVDV, and BoHV-1) responsible for BRD in the field samples. For that purpose, a total of 193 samples (133 nasal swabs and 60 lung tissue samples) from cattle with respiratory signs in northwestern Turkey were examined. For VI, all the samples were inoculated at least 3 blind passages onto MDBK cell culture. In addition, the samples were tested by hemadsorption assay and RT-PCR for BPIV-3; nested RT-PCR for BRSV; immunoperoxidase monolayer assay, antigen-ELISA, and RT-PCR for BVDV; and antigen-ELISA and PCR for BoHV-1. The detected 1 (0.52%) BPIV-3 isolate was found to be in the genotype BPIV-3c. No BRSV isolate could be obtained, while 5 (2.59%) samples were evaluated positive in nested-RT PCR. The presence of BVDV antigen in 10 (5.18%) samples and the BVDV genome in 5 (2.59%) samples were detected, while non-cytopathogenic BVDV isolates were obtained only in 2 (1.04%) samples. The detected BVDV strains fell into the genetic clusters of BVDV-1a, -1f, and -1l. For detection of BoHV-1, although viral isolation and Ag-ELISA results were negative, presence of BoHV-1.1 genome was detected in 2 (1.04%) samples. By the results of VI, ELISA, and PCRs, 10.88% (21/193) of samples were found positive for the evaluated viruses. Depending on the obtained data, combined uses of the diagnostic methods were evaluated to be more reliable for routine diagnosis of bovine respiratory viruses.

The role of the veterinary diagnostic lab in the management of BRD.

Veterinary diagnostic labs (VDLs) are important service agencies providing essential diagnostic testing for a wide variety of domestic animal species as well as wildlife. They serve key roles in disease monitoring and diagnosis as well as surveillance for diseases of consequence. Of the many roles VDLs serve, one is being a member of the larger team of professionals dealing with the management of the bovine respiratory disease (BRD) complex. VDLs provide a number of services related to the management of BRD. These include disease outbreak investigation, abnormal morbidity characterization, routine monitoring, and biosecurity screening for a variety of infectious agents via methods such as necropsy and histopathology, bacterial culture, antimicrobial sensitivity testing, virus isolation, and serological assays. VDLs continue to look for better methods and assays as instrumentation technology also grows and improves. This is reflected in the growing proliferation of molecular-based assays that provide a high degree of sensitivity and specificity. Professional staff in VDLs work in collaboration with those in academia and private industry to conduct basic research focusing on a different aspect of the BRD complex. VDLs remain a primary source of the varied field-origin infectious agents associated with BRD that are used for research purposes.

Behavior assessment and applications for BRD diagnosis: preweaned dairy calves.

Bovine respiratory disease (BRD) is an important disease in dairy calves due to its long-lasting effects. Early identification results in better outcomes for the animal, but producers struggle to identify all calves with BRD. Sickness behavior, or the behavioral changes that accompany illness, has been investigated for its usefulness as a disease detection tool. Behavioral changes associated with BRD include decreased milk intake and drinking speed, depressed attitude, and less likelihood of approaching a novel object or stationary human. Behavioral measurements are useful, as they can be collected automatically or with little financial input. However, one limitation of many BRD behavioral studies includes the use of either lung auscultation or clinical signs as reference methods, which are imperfect. Additionally, external factors may influence the expression of sickness behavior, which can affect if and when behavior can be used to identify calves with BRD. Behavioral measures available to detect BRD lack adequate sensitivity and specificity to be the sole means of disease detection, especially when detection tools, such as calf lung ultrasound, have better test characteristics. However, using behavioral assessments in addition to other detection methods can allow for a robust BRD detection program that can ameliorate the consequences of BRD.

Development and application of molecular diagnostics and proteomics to bovine respiratory disease (BRD).

Advances in molecular and proteomic technologies and methods have enabled new diagnostic tools for bovine respiratory pathogens that are high-throughput, rapid, and extremely sensitive. Classically, diagnostic testing for these pathogens required culture-based approaches that required days to weeks and highly trained technical staff to conduct. However, new advances such as multiplex hydrolysis probe-based real-time PCR technology have enabled enhanced and rapid detection of bovine respiratory disease (BRD) pathogens in a variety of clinical specimens. These tools provide many advantages and have shown superiority over culture for co-infections/co-detections where multiple pathogens are present. Additionally, the integration of matrix-assisted laser desorption ionization time of flight mass spectrometry (MS) into veterinary diagnostic labs has revolutionized the ability to rapidly identify bacterial pathogens associated with BRD. Recent applications of this technology include the ability to type these opportunistic pathogens to the sub-species level (specifically Mannheimia haemolytica) using MS-based biomarkers, to allow for the identification of bacterial genotypes associated with BRD versus genotypes that are more likely to be commensal in nature.

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.

Growth performance and hematological changes of weaned beef calves diagnosed with respiratory disease using respiratory scoring and thoracic ultrasonography.

This study investigated 1) the effect of clinical bovine respiratory disease (BRD) and associated lung consolidations on growth performance and hematological profiles of recently weaned beef calves and 2) the relationship between clinical respiratory signs and lung consolidation detected by thoracic ultrasonography (TUS). One hundred and fifty-three weaned beef calves (209 days old [SD: 35.8] and 306 kg [SD: 26.3], at arrival) purchased and transported from auction markets were accommodated indoors in concrete slatted floor pens. Calves were weighed weekly from arrival until day 28 and on day 65 post-arrival. Assessment of BRD and blood sample collection for hematological profiles were performed on scheduled days (at arrival, on days 7, 14, and 28) and on other days upon BRD diagnosis. Animals were assessed for BRD using a total clinical respiratory score (CRS) of five clinical signs (rectal temperature, ear position, cough, nasal secretion, and eye secretion with each ranging from normal [0] to abnormal [3]) and TUS scores (normal [0] to lung consolidation ≥ 1 cm2 [2]). Based on CRS, 35% of calves were CRS+ (CRS ≥ 5) and 65% were CRS- (CRS < 5). Although no lung consolidations (TUS-) were detected at arrival, 34% of calves developed lung consolidation (≥1 cm2) (TUS+) during the first 28 d post-arrival. Only fever (>39.6 °C) and nasal discharge were weakly associated (r = 0.19, P <0.05) with lung consolidation. On the day of BRD detection, neutrophil number and neutrophil:lymphocyte ratio were 58% and 73% greater, respectively, in BRD calves with lung consolidation compared with healthy calves. From day 0 to 65, calf average daily gain (ADG) did not differ (P >0.05) between CRS+ and CRS- calves but was 0.09 kg/d lower (P < 0.05) for TUS+ compared with TUS- calves. Calves classified as BRD (CRS + TUS ≥ 5) with lung consolidation had lower (P < 0.05) ADG from arrival until day 28 than healthy calves and BRD calves without lung consolidation (0.11 ± 0.10 vs. 0.53 ± 0.07 vs. 0.57 ± 0.10 kg/d, respectively); however, no differences in ADG were observed from day 0 to 65. Conventional methods to diagnose BRD failed to detect calves with lung lesions. TUS is a useful tool to detect lung lesions and its implementation in combination with CRS should provide a more accurate and early diagnosis of BRD, which is fundamental to successful treatment, animal welfare, and growth performance.