Source: COLORADO STATE UNIVERSITY submitted to NRP
METABOLISM AND INFLAMMATION PREDICT CARDIOPULMONARY DISEASE OUTCOMES IN FATTENING BEEF CATTLE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1015583
Grant No.
2018-67015-28241
Cumulative Award Amt.
$499,772.00
Proposal No.
2017-05748
Multistate No.
(N/A)
Project Start Date
Jul 15, 2018
Project End Date
Jul 14, 2023
Grant Year
2018
Program Code
[A1221]- Animal Health and Production and Animal Products: Animal Health and Disease
Recipient Organization
COLORADO STATE UNIVERSITY
(N/A)
FORT COLLINS,CO 80523
Performing Department
Animal Sciences
Non Technical Summary
Profits from beef production in Great Plains feedlots are increasingly offset by health issues. In the past decade, it was observed that fat-cattle can suffer from pulmonary hypertension, a buildup in blood pressure between the heart and lungs. Also, it is very concerning that this condition may be made worse if an animal contracts a respiratory disease, especially late in the fattening phase. The project will focus study on fattening Angus steers, with and without pulmonary hypertension, and then evaluate the impact of viral infection in the cattle suffering from the condition. More specifically, these studies will evaluate how high-pressure between the heart and lungs in fattening steers influences heart and lung tissues, the immune system, and overall behavior; specifically, electronic data systems will record activity and feed intake of healthy and cattle suffering from high-pressure between the heart and lungs and infected with a respiratory virus.Pulmonary hypertension (PH) appears to be responsible for fattening-induced changes in the left-side of the heart, which is a slightly different outcome than observed in cattle suffering from lack of oxygen because of high mountain (i.e., brisket-right heart) disease. We hypothesize that inflammation combined with elevated metabolic demand from fattening contribute to low oxygen in blood and development of PH in feedlot cattle at low elevation. Using fattened Angus steers weaned from low elevation cow-calf operations, we will execute these Specific Aims: 1) utilize marker molecules in blood to evaluate the consequences of PH; 2) utilize these marker molecules to assess inflammation and low oxygen delivery in heart and lungs; and 3) Determine the influence of PH on (a) cattle behavior and performance characteristics, and (b) the response to experimental infection with bovine respiratory syncytial virus, a common pathogen of cattle that causes respiratory disease. This research will identify blood markers that predict disease risk to benefit selective breeding, pre-conditioning, and fattening programs, leading to improved cattle health.
Animal Health Component
33%
Research Effort Categories
Basic
34%
Applied
33%
Developmental
33%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31133101090100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3310 - Beef cattle, live animal;

Field Of Science
1090 - Immunology;
Goals / Objectives
The overall goal of the research described in this proposal is to understand the mechanisms of pulmonary hypertensionin fattening beef cattle, and to use this information to construct tools to help identify at-risk cattle and to develop prevention and new management strategies. This goal is most important if these cattle are exposed to challenges of respiratory disease, such as Bovine Respiratory Syncytial Virus (BRSV)as they finish in thefeedlot.The goals of this project will be accomplished by testing the following hypothesis and executing three specific aims: activation of innate immunity via toll-like receptors (TLR-2,3,4 and TNFRSF1A), and recruitment of circulating inflammatory cells to the heart and lungs mediated by chemochines and receptors (CCL2 and CCR5), in the setting of elevated metabolic demand caused by aggressive fattening regimens in beef cattle, contribute to systemic hypoxemia and are prognostic indicators of worsened PH and adverse outcomes including reduced feed intake and feedlot performance, response to bovine respiratory infection, and pathophysiologic cardiopulmonary remodeling. The following Specific Aims are enumerated.1. To utilize blood biomarkers of systemic hypoxemia, metabolism, and innate immune activation to prospectively evaluate the consequences of PH in fattening beef cattle.2. To utilize the biomarkers described above to assess inflammatory cell recruitment, metabolic status, and hypoxia-induced gene expression in post-mortem target cardiopulmonary tissues.3. To determine the influence of PH on (a) cattle behavior and performance characteristics, and (b) the response to infection with bovine respiratory disease.
Project Methods
Angus-based steers (n = 300) weaned from low elevation cow-calf operations will be placed in the Eastern Colorado Research Center(1,400 m) feedlot. These steers will be procured from operations from the Great Plains (350 to 1,600 m elevation), which have history of heart failureand have not been selectively bred for low PAP (pulmonary arterial pressue is a tool that ids cattle at risk of pulmonary hypertension in high altitude). Steers will initially weigh 275-300 kg. We expect to feed the steers for 205 days and harvest them when they weigh 600 kg. Feeding of steers will progress thru step-up rations and a growing-finishing ration of ~80% concentrate and ~20% roughage throughout this study as to gain ~1.5 kg/day.The study will be a 2 x 2 factorial design with High vs Low PAP as the first level and viral challenge (i.e., Vehicle control (Vhc) vs bovine respiratory syncytial virus (BRSV)) as the second. The final 70 days of finishing of these steers will be managedin the feed intake unit at CSU's Agricultural Research and Development Education Center (ARDEC) near Fort Collins to facilitate intensive data collection and cattle handling. The following bullet points and times (day) describe the steer management procedures of the study. Day 0: Steers received Eastern Colorado Research Centerfeedlot. Initial processing will include vaccinations and all other health management - standard operating procedures of this research facility. If a steer is pulled from their pen for health complications, that animal will be excluded from the study.Day 60: After 60 days of feeding, blood will be collected and PAP measured. Fifty highest and 50 lowest mPAP steers identified. Steers expected to weigh ~360 kg at this time.Day 130: To verify High and Low PAP status, the 100 steers will receive second PAP measurement and blood collected. Steers expected to weigh 500 kg. Steers with the highest (n = 20) and lowest (n = 20) PAP will be moved to the feed intake facility at ARDEC.Day 135: Once received at ARDEC, steers will be equipped with intensive data monitoring devices for health, activity, and feed intake traits. Once in this facility, steers will be weighed and ultrasound scanned for carcass traits every two weeks. Steers are expected to be in this facility for at least 70 days before carcass and tissue harvest.Days 195 to 204: mPAP measured and blood collected and the following procedures executed:Day 195: Challenged with saline Vehicle (Vhc) or BRSV.Day 198: Blood collectionDay 201: Blood collectionDay 204: Blood collection and mPAP measurement.Days 205 to 210: Steers slaughtered and cardiopulmonary tissues harvested. The 24 steers that best fit the categories of the 2 x 2 factorial design will be used forintensive laboratory analyses.Steers will be slaughtered at a cooperating packing plant, Innovative Foods, Inc. (Evans, CO), where organs and tissues will be collected and processed for analysis. Staff of ARDEC will monitor steers twice per day, and these efforts will be greatly assisted with the two electronic data collection systems.The following laboratory, tissue and whole animal data collection/analyses will be conducted to as to understand the influnece of PAP level and virus challenge on steer health:Circulating biomarkers. IL-6 (as presented in Figure 5), TNFα, and MCP-1 will be measured by ELISA (Kingfisher Biotech) as accepted indices of inflammatory status in PH. Serum uric acid, known as a biomarker of metabolic and inflammatory stress in a number of disease states, and as a prognostic indicator of PH, will be measured by enzyme assay (Hoeper et al., 1999a). Brain natriuretic peptide (BNP) and its pro-peptide amino-terminal pro BNP (NT-proBNP) are accepted indicators of cardiac dysfunction in human PH and will be measured by commercial ELISA. The BNP peptide is conserved across species and NT-proBNP will be measured with a bovine specific ELISA. Additional standard serum measurements will include pH and electrolytes, creatinine, bilirubin, and liver enzymes. Several of these, including elevated serum uric acid and creatinine, hypoferremia, and hyponatremia, have prognostic ability in humans with PH (Robinson et al., 2014) Additional analyes ofinnate immunity using RNA from peripheral blood mononuclear cells will also be executed the targets of these analyses include:TLR-2,3,4, TNFRSF1A, CCL2, and CCR5.Metabolome analysis. To assess the influence of high intensity fattening on metabolic re-programming, we will perform ultrahigh performance liquid chromatography-mass spectrometry at the UC-Denver Metabolomics Core Lab. In brief, plasma aliquots will be separated chromatographically and quantitated by mass spectroscopy. Metabolite assignment and quantitation will be calculated with the software Maven (Princeton) against the KEGG pathway database and in-house validated library of > 650 compounds. These results will be compared between groups while correcting for false discovery rate using Benajamini-Hochberg algorithms for all significant metabolites. Supervised partial least-square discriminant analysis (Simca, Umetrics, San Jose CA) will then be used to determine the variance of metabolic phenotypes between groups. This analysis will provide an unprecedented depth of data on the integrated metabolic adaptations to fattening from consumption of a high caloric diet and mechanistic insights to the metabolic bases of disease.Histopathology: Pathologic remodeling associated with altered expression of blood biomarkers, samples of lung, right and left ventriclefree wall, moderator band, interventricular septum, right and left atrium, pulmonary artery, aorta, liver, longissiumus dorsi muscle, and major organ systems along with any macroscopic lesions will be collected for formalin fixation and processed for histopathology. Sections will be stained with hematoxylin and eosin (H&E) and selected sections of lung, heart and the great vessels will be stained to demonstrate collagen and elastin (Masson's Trichrome, Verhoeff-Van Gieson), and mineralization (Von Kossa). The severity of lung and heart lesions will be blind scored in blind scored using a grading system to assess pulmonary vascular remodeling, cardiomyocyte hypertrophy and replacement fibrosis. Lungand heart morphology will also be executed with these tissues andH&E sectionsscanned withthe Olympus VS110 Virtual Slide Scanner and image acquisition software (Cell Sens).Animal performance and behavior. When steers are placed in the feed intake system (Grow-Safe®; http://www.growsafe.com) at ARDEC approximately 70 days before slaughter, the 40 forty steers will also be fitted with the ear-tag apparatus of the CowManager® (https://www.cowmanager.com/en-us/). These biomonitoring systems will record feed intake, movement/activity, rumination, and body temperature in real-time. Summaries of data will be downloaded daily, and most importantly, if there is a spike in body temperature above the norm, the system will notify immediately via phone-app. Body weights will be obtained every 2 weeks and steers will be ultrasound scanned to measure carcass traits at these times.

Progress 07/15/18 to 07/14/23

Outputs
Target Audience:Scientists working in the disciplines of animal agriculture, veterinary health of feedlot cattle, and beef industry, specifically the feedlot segment. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project provided the resources for the training of two postdoctoral fellows. The project did not pay the salaries of the two trainees, but provided animals, samples, data and opportunity to analyze data and write papers. The trainees included, Drs. Greta M. Krafsur and Miranda M. Culbertson. Dr. Krafsur is a DVM that was also pursuing a PhD in the Department of Microbiology, Immunology, and Pathology in the Colorado State University College of Veterinary Medicine and Biomedical Sciences. During the period of this grant, Dr. Krafsur also became a board-certified pathologist and now works in the biomedical field as a diagnostic pathologist. Dr. Culbertson was trained in the discipline of Breeding and Genetics at Colorado State University and this project provided her opportunity to learn about the feedlot and harvest industry, management of research cattle, and about data analysis and manuscript writing. Dr. Culbertson is now Beef Extension Specialist at Iowa State University. How have the results been disseminated to communities of interest?Information was translated to the animal agriculture scientific community and the beef industry by the following written publication and presentations. Peer-reviewed journals: C. Zhai, L.C. Li Puma, A.J. Chicco, A. Omar, R.J. Delmore, I. Geonaras, S.E. Speidel, T.N. Holt, M.G. Thomas, R.M. Enns, and M.N. Nair. 2022. Pulmonary arterial pressure in fattened Angus steers at moderate altitude influences early postmortem mitochondria functionality and meat color during retail display. J. Anim. 100:1-9. Doi.org/10.1093/jas/skac002. Manuscripts in review or in preparation for submission to peer-review journals: G.M. Krafsur, R.M. Enns, M.G. Thomas, T.N. Holt, G.L. Mason, D.H. Gould, R.D. Brown, and K.R. Stenmark. 2023. Post-harvest heart score as a surrogate marker for adverse cardiopulmonary remodeling in fattened beef cattle with pulmonary hypertension and congestive heart failure. To be submitted J. Anim. Sci. 2023. M.M. Culbertson, G.M. Krafsur, R.D. Brown, T.N. Holt, S.E. Speidel, R.M. Enns, R.A. Bowen, R.J. Delmore, K.R. Stenmark, and M.G. Thomas. 2023. Feedlot performance, pre-harvest blood parameters, and carcass and heart characteristics among fattening Angus-influenced beef steers sorted into high and low pulmonary arterial pressure groups. To be submitted: J. Anim. Sci. 2023. Abstracts: C. Zhai, L.C. Li Puma, A.J Chicco, A. Omar, R.J. Delmore, I. Geornaras, S.E. Speidel, T.N. Holt, M.G. Thomas, R.M. Enns, and M.N. Nair. High pulmonary arterial pressure in steers at moderate altitude affects early postmortem mitochondria functionality. Am. Meat Sci. Assoc. Ann. Recip. Meat Conf., August 15 - 18, 2021, Reno, NV., Abstract #123. G. Krafsur, R. Brown, M. Thomas, M. Culbertson, T. Holt, S. Speidel, R. Enns, K. Stenmark, M. Li, S. Riddle, and R. Bowen. 2020. Metabolism and inflammation predict cardiopulmonary disease outcomes in fattening beef cattle: histopathology. Conf. Res. Workers Anim. Disease. Virtual. Abstr. 284. C. Zhai, R.J. Delmore, I. Geonaras, S. Speidel, T. Holt, M. Thomas, M. Enns, and M.N. Nair. 2020. Pulmonary arterial pressure in cattle influences beef color during retail display. 2020. Proc. Joint Mtg. ASAS-CSAS-WSASAS (AS = Animal Science). Virtual, July 19-23, 2020. Abstr. PSV-32. G.M. Krafsur, R. Brown, T.N. Holt, D.H. Gould, F. Garry, S. Riddle, J. Neary, M. Enns, M. Thomas, and K.R. Stenmark. 2018. Biomarkers of inflammation and metabolic stress in a bovine model of pulmonary hypertension with left ventricular fibrosis and pulmonary venous remodeling. Am. J. Crit. Care Med. 197:A3748 Invited Academic-Scientific-Technical Presentations: Heart failure in beef cattle - PAP EPD development. International Genetic Solutions (IGS) booth seminar at National Cattlemen's Beef Association meeting, San Antonio, TX February 5, 2020. Mid-late feeding mortalities in finishing cattle and potential causes. Zoetis WebEx for Global Technical Services and Veterinary Medical Research and Development. November, 6, 2019. Late feeding mortality and pulmonary hypertension. Think-tank for "Late Day Morbidity Challenges in High Performance Cattle." Proc. Published by Noble Foundation, Ardmore, OK, September 19, 2019. p. 49-57. PAP, HMD, FHD: alphabet soup of what we know and don't know about genetics of pulmonary hypertension in cattle. BIF Symposium: Advancements in efficiency and adaptability. Brookings, SD, June 19, 2019. Mid-late feeding mortalities in finishing cattle and potential causes. Round-table Discussions of Academy of Veterinary Consultants, sponsored by 5-Rivers Cattle Feeding, LLC. Loveland, CO, August 7, 2019. Mid-late feeding mortalities in finishing cattle and potential causes. Micronutrients Global Species Council Meeting, Fort Collins, CO, June 5, 2019. Mid-late feeding mortalities in finishing cattle and potential causes. Midwest PMS Consultant meeting, Denver, CO, May 7, 2019. Mid-late feeding mortalities in finishing cattle and potential causes. Plains Nutrition Conference. San Antonio TX, April 12, 2019. Mid to late feeding mortalities in finishing cattle and potential causes. High Plains Beef Symposium - American Society of Animal Sciences. Austin, TX, July, 11, 2019. Extension Presentations: Ongoing research update on pulmonary hypertension. Pulmonary arterial pressure (PAP) and Heart Scoring Certification for Colorado Veterinarians. Fort Collins, CO, August 12, 2022. Feedlot heart disease: new trait development and validation of packing plant heart score. Beef Improvement Federation Annual Meeting, Las Cruces, NM, June 3, 2022. Cattle HAD, FHD, and PAP; understanding pulmonary hypertension in cattle. National Cattlemen's Beef Association Webinar, Health Issues in Mid-late Fed Cattle. May 5, 2020. Cattle HAD, FHD, and PAP; understanding pulmonary hypertension in cattle. Leachman Cattle of Colorado High Altitude Bull Sale, Loma, CO, December 6, 2019. PAP-an important trait for mountain and feedlot cattle. Leachman Cattle of Colorado Fall Bull Sale Educational Program. Fort Collins, CO. November 8, 2019. Research updates on pulmonary hypertension and feedlot heart disease. Colorado Veterinary Pulmonary Arterial Pressure Summit. Fort Collins, CO. August 3, 2018. Extension Articles, Cattle Grower Reports, and Industry Board Advisory Reports. M. Reiman and M.G. Thomas. 2022. Healthy hearts. Start with knowing more. Bovine congestive heart failure research reveals new information. Angus J. August 2022. H. Smith-Thomas with support of M.G. Thomas. 2020. New Research probes into feedlot cattle heart failure. Progr. Cattle. 8:42-43. M.G. Thomas, G.M. Krafsur, T.N. Holt, R.M. Enns, and S.E. Speidel. 2020. Understanding late day morbidity in fed cattle: focus on pulmonary hypertension (PH). Proceedings fort the Late Day Morbidity Challenges in High Performance Cattle, Industry Think-Tank, Noble Research Institute, Ardmore, OK. September 18-20, 2019. H. Smith-Thomas with support from M.G. Thomas. 2019. Research looks at congestive heart failure in feedlot cattle. SimTalk 27(5):30-32. H. Smith-Thomas with support from M.G. Thomas. 2019. Research looks at congestive heart failure in feedlot cattle. Feed-Lot, Feeder Information Highlights 27(6):14-17. S.E. Speidel, M.G. Thomas, and R.M. Enns. 2019. Pulmonary arterial pressure and their utility for cow-calf producers. Proceedings, Range Beef Cow Symposium XXVI, Scotts Bluff, NE. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Twenty-nine communications translated results of this project to the animal agriculture scientific community and beef industry. Two post-doctoral fellows also received training. The primary findings of this project included, but not limited to, 1) Angus cattle (i.e., calf-feds) in this project were suffering from PH after 3 months of being placed in the feedlot, 2) tools that measure PH, such as PAP and blood gases (oxyhemoglobin concentration), revealed hypoxia-differences among steers of high (H) and (L) PAP categories, but the cattle had minimal differences in their leukogram prior to harvest despite gene expression pathway (Complement and Acute Phase Inflammatory Response and oxidative stress) differences observed in RNA-seq data from peripheral blood mononucleocytes, 3) viral challenge at the end of the feeding period had limited influence on these steers, 4) HPAP relative LPAP steers had poor average daily gain and feed efficiency, even though cattle had similar body and carcass weights and carcass trait measures, and 5) steaks from HPAP steers had one day less shelf life than steaks from LPAP, and 6) preliminary results from this project supported accrual of much larger funding from the Foundation for Food and Agriculture Research - International Consortium for Antimicrobial Stewardship in Agriculture (FFAR-ICASA; > $ 1 million for two projects) related to liver abscess and heart failure in feedlot cattle. Overall conclusions of this project are that death-loss in these steers was 5.5% and appeared to be a result that the Angus steers withHPAP experienced PH through a 9-month growing and fattening period. Most importantly, steers harvested from the HPAP grouping had cardiac histomorphologic alterations (i.e., elevated heart scores). The observations that the steers with high PAP early in the feeding period suggested that late feedlot death is a consequence of the early increases in PAP and PH in the feeding period, but the detrimental phenotypes are likely not observed until the end of the feeding period. The sampling of cattle for biomarkers at the end of the feeding period did not appear to be informative of the early PH response in the feeding period; however, additional research is needed to validate how early feedlot entry parameters predict later feedlot performance and health phenotypes. Below are summaries of the multi-disciplinary accomplishments of this project. Development of a heart scoring system: Adverse cardiopulmonary remodeling underlies elevated PAP, PH and right ventricular (RV) failure. Morphologic alterations in the cardiac RV are visualized as per the phenotypic characterization of heart score methodology (HS; 1-5). We hypothesized that post-harvest HS tracked with histomorphologic alterations in bovine cardiopulmonary explants obtained from fed cattle with PAP data, increasing confidence in heart score as an indicator for feedlot PH and CHF. Forty-two Angus steers (which included steers from this NIFA project) with PAP data were assigned heart scores at harvest, ventricular mass was determined, and cardiopulmonary tissues were excised for histopathology analyses. Heart score, was moderately and positively correlated with PAP (r ? 0.50; P < 0.05) and ventricular RV, and left ventricle and septum weight. Heart score was moderately also associated (r ? 0.50; P < 0.05) with cardiac fibrosis levels in the interstitial and perivascular compartments and indicators in the RV and LV interstitial and perivasculature. Feedlot performance, pre-harvest blood parameters, and carcass and heart characteristics among high and low PAP Angus steers: Using fattening steers (n = 107) from a moderate (1250 m) elevation cow-calf operation with a history of feedlot heart disease, Angus-influenced steers were assigned in a 2 x 2 factorial design to groups of PAP, and viral challenge (control versus bovine respiratory syncytial virus (BRSV)). Post-weaning, steers were fed a growth-finishing ration targeting gain of 1.5 kg/day for 9 months. The average PAP for cattle on feed was 44.7 ? 1.3, 52.3 ? 1.9, 65.1 ? 4.0 mm Hg at 3, 6, and 9 months of feeding, respectively. Steers determined to be either high (HPAP; n = 23) or low (LPAP; n = 22) PAP were placed in a feed intake evaluation system for the final 60 days of finishing. Final PAP measurement was 11 days prior to harvest with means for the LPAP and HPAP groups of 44.1 ? 4.3 mm Hg and 87.2 ? 4.3 mm Hg (P < 0.01), respectively. Consequently, HPAP steers were susceptible to heart failure. Death loss in this study was 5.5% with all mortalities attributed to PH-induced right heart failure. These mortalities occurred after 6 months of feeding. Final body weight (595.8 ? 14.0 kg) and carcass traits were similar among the two groups and BRSV challenge had no affect on death loss, carcass traits, heart weights and heart scores. The LPAP steers had superior (P < 0.04) ADG and feed efficiency than the HPAP steers. Venous blood gases unveiled hypoxemia in both groups; however, respiratory acidosis, hypercapnia, was also observed in the HPAP steers and oxyhemoglobin concentration was considerably decreased (P < 0.01) in HPAP steers relative the LPAP steers. Neither group had an inflammatory leukogram. The HPAP steers had elevated heart scores (P < 0.5) arising from increased mass of the RV. Gene expression in peripheral blood mononucleocytes (PBMC): Gene expression of innate immunity via toll-like receptors (TLR- 2,3,4 and TNFRSF1A), and recruitment of circulating inflammatory cells to the heart and lungs mediated by chemokines and receptors (CCL2 and CCR5) were elevated in the setting of elevated metabolic demand caused by fattening of Angus steers. We hypothesized that inflammatory activation of peripheral blood mononuclear cells (PBMC) accompanies PH in affected cattle. Transcriptome profiling with RNA-Seq was performed using PBMC isolated from HPAP (n=22) and LPAP (n=21) steers at the termination of fattening. Informatic analysis showed that PBMC from PH-affected cattle exhibited gene expression signatures consistent with immune-inflammatory cell activation and mobilization for recruitment from the circulation and homing/migration to the periphery. A subset of 35 genes was identified that distinguish PH-affected and unaffected steers. These results provided candidate blood biomarkers for early and minimally invasive detection of animals at risk; however, these approaches need validation. Since there were blood samples collected at the 3-month sampling, Dr. Stenmark's team will be investing funds from NIH programs to evaluate blood biomarkers and their predictability of hypoxia-related health parameters as animals fatten to obese states. Impacts of feedlot hypoxia on meat quality: PAP is an indicator for PH in cattle. The impact of pulmonary arterial pressure (PAP) on mitochondrial function, oxidative phosphorylation (OXPHOS) protein abundance, and meat color was examined using longissimus lumborum (LL) from HPAP and LPAP steers during early postmortem period (2 h and 48 h) and retail display (day 1 to 9), respectively. High PAP muscle had greater (P < 0.05) OXPHOS-linked respiration and proton leak than low PAP muscles at 2 h postmortem but rapidly declined to be similar (P > 0.05) to low PAP muscle by 48 h postmortem. OXPHOS protein expression was higher (P < 0.05) in low PAP than high PAP muscle. During retail display, redness, chroma, hue, ratio of reflectance at 630 nm and 580 nm, and metmyoglobin reducing activity decreased faster (P < 0.05) in steaks from HPAP relative LPAP steers. Lipid oxidation increased (P < 0.05) in steaks from HPAP but not (P > 0.05) LPAP. The results suggested that high PAP caused a lower OXPHOS efficiency and a greater fuel oxidation rate under conditions of low ATP demand in premortem beef LL muscle, this could explain the lower feed efficiency in high PAP feedlot cattle.

Publications

  • Type: Journal Articles Status: Published Year Published: 2022 Citation: C. Zhai, L.C. Li Puma, A.J. Chicco, A. Omar, R.J. Delmore, I. Geonaras, S.E. Speidel, T.N. Holt, M.G. Thomas, R.M. Enns, and M.N. Nair. 2022. Pulmonary arterial pressure in fattened Angus steers at moderate altitude influences early postmortem mitochondria functionality and meat color during retail display. J. Anim. 100:1-9. Doi.org/10.1093/jas/skac002.
  • Type: Journal Articles Status: Other Year Published: 2023 Citation: G.M. Krafsur, R.M. Enns, M.G. Thomas, T.N. Holt, G.L. Mason, D.H. Gould, R.D. Brown, and K.R. Stenmark. 2023. Post-harvest heart score as a surrogate marker for adverse cardiopulmonary remodeling in fattened beef cattle with pulmonary hypertension and congestive heart failure. To be submitted J. Anim. Sci. 2023.
  • Type: Journal Articles Status: Other Year Published: 2023 Citation: M.M. Culbertson, G.M. Krafsur, R.D. Brown, T.N. Holt, S.E. Speidel, R.M. Enns, R.A. Bowen, R.J. Delmore, K.R. Stenmark, and M.G. Thomas. 2023. Feedlot performance, pre-harvest blood parameters, and carcass and heart characteristics among fattening Angus-influenced beef steers sorted into high and low pulmonary arterial pressure groups. To be submitted: J. Anim. Sci. 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: C. Zhai, L.C. Li Puma, A.J Chicco, A. Omar, R.J. Delmore, I. Geornaras, S.E. Speidel, T.N. Holt, M.G. Thomas, R.M. Enns, and M.N. Nair. High pulmonary arterial pressure in steers at moderate altitude affects early postmortem mitochondria functionality. Am. Meat Sci. Assoc. Ann. Recip. Meat Conf., August 15  18, 2021, Reno, NV., Abstract #123.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: G. Krafsur, R. Brown, M. Thomas, M. Culbertson, T. Holt, S. Speidel, R. Enns, K. Stenmark, M. Li, S. Riddle, and R. Bowen. 2020. Metabolism and inflammation predict cardiopulmonary disease outcomes in fattening beef cattle: histopathology. Conf. Res. Workers Anim. Disease. Virtual. Abstr. 284.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: C. Zhai, R.J. Delmore, I. Geonaras, S. Speidel, T. Holt, M. Thomas, M. Enns, and M.N. Nair. 2020. Pulmonary arterial pressure in cattle influences beef color during retail display. 2020. Proc. Joint Mtg. ASAS-CSAS-WSASAS (AS = Animal Science). Virtual, July 19-23, 2020. Abstr. PSV-32.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: G.M. Krafsur, R. Brown, T.N. Holt, D.H. Gould, F. Garry, S. Riddle, J. Neary, M. Enns, M. Thomas, and K.R. Stenmark. 2018. Biomarkers of inflammation and metabolic stress in a bovine model of pulmonary hypertension with left ventricular fibrosis and pulmonary venous remodeling. Am. J. Crit. Care Med. 197:A3748


Progress 07/15/21 to 07/14/22

Outputs
Target Audience:National and International Scientific communities with interests in beef cattleas well as the U.S. beef industry. Changes/Problems:We cut and paste the information from the last box also into this box; therefore, We are asking for a twelve-month no cost extension (July 14-2022 to July 14-2023). This time frame will primarily be used to complete manuscripts and pay for their page charges and conduct further analyses to maximize the results. The draft titles of manuscripts are listed below. While navigating the challenges of the pandemic, we have been able to complete analyses of animal performance and health, histopathology analyses of the tissues from the study animals, and most recently, analyze gene expression data obtained from RNA-Sequence of the peripheral blood mononucleocytes (PBMC) obtained from healthy steers versus steers suffering from pulmonary hypertension. These samples were collected a few days prior to finished harvest of the animals (i.e., ~600 kb body weight and 15 months of age). Specifically, these were fattening/finished Angus steers and the gene expression results is our most recent accomplishment. The initial review of these results suggest that the consequences of pulmonary hypertension was present in the high PAP (pulmonary arterial pressure) categorized steers when they entered the feeding phase of their life (i.e., 7 months of age and ~320 kg of body weight). This is an important finding as our initial hypotheses suggested that feedlot heart disease, and potentially late feedlot death, was an etiology from inflammation signatures that developed late in the feeding and fattening phase. Even though this hypothesis may be supported in some groups of cattle in U.S. feedlots, the animals of this study may be of another disease sub-group that develop pulmonary hypertension at a young age (i.e., pre-weaning) and therefore, experience a life-long cascade of a diseased cardio-pulmonary system that leads to poor feedlot performance and potentially late feedlot death. These new findings warrant our team of scientist additional time to consider the new and novel results as these findings are potentially impactful to challenge of feedlot heart disease being observed in the U.S. cattle feeding segment of the beef industry. These data may also be insightful to the biomedical community as fattening steers are a strong animal model for understanding consequences of high caloric intake the ontogeny of increasing levels of adiposity and subsequent cardio-pulmonary malformations. Manuscripts close to submission: G.M. Krafsur, R.M. Enns, M.G. Thomas, T.N. Holt, G.L. Mason, D.H. Gould, R.D. Brown, and K.R. Stenmark. 2022. Post-harvest heart score as a surrogate marker of adverse cardiopulmonary remodeling in fattened beef cattle with pulmonary hypertension and congestive heart failure. Target journal = Journal of Animal Science. M.G. Thomas, T.N. Holt, M.M. Culbertson, R.D. Brown, G.M. Krafsur, S.E. Speidel, R.M. Enns, R.A. Brown, and K.R. Stenmark. 2022. Feedlot performance, carcass characteristics, and blood parameters among fattening Angus beef steers sorted into high and low pulmonary arterial pressure groups. Target journal = Journal of Animal Science. Manuscripts in the early preparation phase: G.M. Krafsur et al. 2022. Histopathology comparisons of cardiopulmonary tissues harvested from fattened Angus sorted into high and low pulmonary arterial pressure groups. Target journal = to be determined. K.R. Stenmark et al., 2022. Inflammation and tissue remodeling signatures observed in RNA-Seq gene expression data from peripheral blood mononucleocytes harvested from fattened Angus steers sorted into high and low pulmonary arterial pressure groups: early life biomarker prediction of later life pulmonary hypertension phenotypes. Target journal: to be determined, expecting submission to high impact factor journal spanning biomedical and animal physiology readership. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Results were disseminated to scientific communities through manuscripts and abstract presentation. Most importantly, gave an invited presentation at the Beef Improvement Federation meeting as to help the beef industry further understand pulmonary hypertension and late feedlot death. Below is the title of that presentation: Feedlot heart disease: new trait development and validation of packing plant heart score. Beef Improvement Federation Annual Meeting, Las Cruces, NM, June 3, 2022. What do you plan to do during the next reporting period to accomplish the goals?We are currently asking for a twelve-month no cost extension to this project (July 14-2022 to July 14-2023). This time frame will primarily be used to complete manuscripts and pay for their page charges and conduct further analyses to maximize the results. The draft titles of manuscripts are listed below. While navigating the challenges of the pandemic, we have been able to complete analyses of animal performance and health, histopathology analyses of the tissues from the study animals, and most recently, analyze gene expression data obtained from RNA-Sequence of the peripheral blood mononucleocytes (PBMC) obtained from healthy steers versus steers suffering from pulmonary hypertension. These samples were collected a few days prior to finished harvest of the animals (i.e., ~600 kb body weight and 15 months of age). Specifically, these were fattening/finished Angus steers and the gene expression results is our most recent accomplishment. The initial review of these results suggest that the consequences of pulmonary hypertension was present in the high PAP (pulmonary arterial pressure) categorized steers when they entered the feeding phase of their life (i.e., 7 months of age and ~320 kg of body weight). This is an important finding as our initial hypotheses suggested that feedlot heart disease, and potentially late feedlot death, was an etiology from inflammation signatures that developed late in the feeding and fattening phase. Even though this hypothesis may be supported in some groups of cattle in U.S. feedlots, the animals of this study may be of another disease sub-group that develop pulmonary hypertension at a young age (i.e., pre-weaning) and therefore, experience a life-long cascade of a diseased cardio-pulmonary system that leads to poor feedlot performance and potentially late feedlot death. These new findings warrant our team of scientist additional time to consider the new and novel results as these findings are potentially impactful to challenge of feedlot heart disease being observed in the U.S. cattle feeding segment of the beef industry. These data may also be insightful to the biomedical community as fattening steers are a strong animal model for understanding consequences of high caloric intake the ontogeny of increasing levels of adiposity and subsequent cardio-pulmonary malformations. Manuscripts close to submission: G.M. Krafsur, R.M. Enns, M.G. Thomas, T.N. Holt, G.L. Mason, D.H. Gould, R.D. Brown, and K.R. Stenmark. 2022. Post-harvest heart score as a surrogate marker of adverse cardiopulmonary remodeling in fattened beef cattle with pulmonary hypertension and congestive heart failure. Target journal = Journal of Animal Science. M.G. Thomas, T.N. Holt, M.M. Culbertson, R.D. Brown, G.M. Krafsur, S.E. Speidel, R.M. Enns, R.A. Brown, and K.R. Stenmark. 2022. Feedlot performance, carcass characteristics, and blood parameters among fattening Angus beef steers sorted into high and low pulmonary arterial pressure groups. Target journal = Journal of Animal Science. Manuscripts in the early preparation phase: G.M. Krafsur et al. 2022. Histopathology comparisons of cardiopulmonary tissues harvested from fattened Angus sorted into high and low pulmonary arterial pressure groups. Target journal = to be determined. K.R. Stenmark et al., 2022. Inflammation and tissue remodeling signatures observed in RNA-Seq gene expression data from peripheral blood mononucleocytes harvested from fattened Angus steers sorted into high and low pulmonary arterial pressure groups: early life biomarker prediction of later life pulmonary hypertension phenotypes. Target journal: to be determined, expecting submission to high impact factor journal spanning biomedical and animal physiology readership.

Impacts
What was accomplished under these goals? 1. To utilize blood biomarkers of systemic hypoxemia, metabolism, and innate immune activation to prospectively evaluate the consequences of PH in fattening beef cattle. 2. To utilize the biomarkers described above to assess inflammatory cell recruitment, metabolic status, and hypoxia-induced gene expression in post-mortem target cardiopulmonary tissues. For numbers 1 and 2, we have completed an RNA-Seq analysis of the peripheral blood mononucleocyte from fattened steers suffering from pulmonary hypertension (i.e., comparison of steers with high vs low pulmonary arterial pressure). Please see below: Gene ontology analyses (Figures A and B below) from RNA harvested from PBMC slightly before fattened/finished harvest of Angus steers suggested activation of pro-inflammatory circulating immune cells and mobilization for recruitment to target organs. The data also confirmed expected increases in candidate genes (Figure C) that were proposed in the initial grant to be reactive to pulmonary hypertension in Angus cattle. Therefore, these new results are encouraging as per their support of the initial thoughts drafting this proposal; however, we would like to have more time to consider these new results and how they may have been influenced by blood parameters in samples that were collected 7 months prior. Note, we could differentiate high and low PAP cattle at the start of this study when the cattle only weighed ~320 kg; therefore, as they doubled in size to ~600 kg of body weight, several suffered long-term from pulmonary hypertension. To further support this request, the RNA-Seq results were also used to generate a receiver operator curve and it had strong area under the curve (0.945), sensitivity of 90.1%, and specificity of 100%; therefore, a subset of genes were associated with were associated with fattening-induced PH phenotype, but gene expression did not correlate quantitatively with elevated mPAP; therefore, suggesting the PBMC in blood expression profile was most likely brought into this study when the cattle were post-weaning. Because of these results were asking for a no cost extension to this project as further need o study relationships between blood biomarkers in the initial samples collected 7 months prior to these samples collected with the cattle were harvested at 15 months of age and ~600 kg of body size. See Figures A-D at the end of this section. 3. To determine the influence of PH on (a) cattle behavior and performance characteristics, and (b) the response to infection with bovine respiratory disease. For this Aim, we collected and have started reporting growth, feed efficiency, carcass, and pulmonary arterials pressure data. In brief, in comparisons of cattle with high versus low pulmonary arterial pressure we found that cattle had similar body weights and carcass traits; however, the high PAP steers had horrible efficiency traits such as average daily gain and feed efficiency. We are currently working to publish these results. We expect to submit a manuscript to Journal of Animal Science within 3 to 4 months. The title of this manuscript is below as is another manuscript that will validate a heart scoring procedure that can be used to collect massive amounts of data for genetic improvement efforts to try and reduce pulmonary hypertension in fattening beef cattle. G.M. Krafsur, R.M. Enns, M.G. Thomas, T.N. Holt, G.L. Mason, D.H. Gould, R.D. Brown, and K.R. Stenmark. 2022. Post-harvest heart score as a surrogate marker of adverse cardiopulmonary remodeling in fattened beef cattle with pulmonary hypertension and congestive heart failure. Target journal = Journal of Animal Science. M.G. Thomas, T.N. Holt, M.M. Culbertson, R.D. Brown, G.M. Krafsur, S.E. Speidel, R.M. Enns, R.A. Brown, and K.R. Stenmark. 2022. Feedlot performance, carcass characteristics, and blood parameters among fattening Angus beef steers sorted into high and low pulmonary arterial pressure groups. Target journal = Journal of Animal Science.

Publications

  • Type: Journal Articles Status: Published Year Published: 2022 Citation: C. Zhai, L.C. Li Puma, A.J. Chicco, A. Omar, R.J. Delmore, I. Geonaras, S.E. Speidel, T.N. Holt, M.G. Thomas, R.M. Enns, and M.N. Nair. 2022. Pulmonary arterial pressure in fattened Angus steers at moderate altitude influences early postmortem mitochondria functionality and meat color during retail display. J. Anim. 100:1-9. Doi.org/10.1093/jas/skac002.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: C. Zhai, L.C. Li Puma, A.J Chicco, A. Omar, R.J. Delmore, I. Geornaras, S.E. Speidel, T.N. Holt, M.G. Thomas, R.M. Enns, and M.N. Nair. High pulmonary arterial pressure in steers at moderate altitude affects early postmortem mitochondria functionality. Am. Meat Sci. Assoc. Ann. Recip. Meat Conf., August 15  18, 2021, Reno, NV., Abstract #123..
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Feedlot heart disease: new trait development and validation of packing plant heart score. Beef Improvement Federation Annual Meeting, Las Cruces, NM, June 3, 2022.


Progress 07/15/20 to 07/14/21

Outputs
Target Audience: Scientist with background in Animal Health and Beef Cattle. Livestock producers with experience in cattle feeding. Changes/Problems:We extended this award for 1 year. This extension was necessary due to limitation in our daily job efforts because of the COVID19 pandemic. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Abstracts was presented at the Conference of Researchers Working in Animal Disease (CRWAD), which is a scientific audience with great interest in feedlot heart disease. Extension efforts were put forth to both cattle breed associations and feedlot nutrition companies and managers. Specifically, data/results were presented to employees of MidWest PMS, LLC which represents approximately 1 million head of beef cattle in feedlots. What do you plan to do during the next reporting period to accomplish the goals?We will continue to analyze data and prepare manuscripts and abstracts.

Impacts
What was accomplished under these goals? In the past year, we completed RNA-Sequencing of the peripheral blood mononeuclecotyes as to start to evaluate gene expression inthese indicative of immune response due to high and low conductions in the pulmonary artery as high PAP (pulmonary arterial pressure) is indicative of Pulmonary Hypertension. We also started to evaluate heart and lung tissues using histopathology techniques and we observed striking infiltration of the cardiac perivascular and interstitial compartments with fibrotic and lymphomononuclear inflammation in the perivascular spaces and cardiac interstitium. Coronary ateriosclerotic lesions were also observed in the left cardiac ventricular free wall and papillary muscle. High PAP steers also had outstanding BALT (bronchus-associated lymphoid tissue) hyperplasia and adverse remodeling of the pulmonary artery and venous circulations. Overall, we are making progress understanding the phenotypes of feedlot heart disease and can use the RNA-seq/gene expression data to start to delineate candidates for blood biomarkers discovery.

Publications

  • Type: Other Status: Published Year Published: 2020 Citation: G. Krafsur, R. Brown, M. Thomas, M. Culbertson, T. Holt, S. Speidel, R. Enns, K. Stenmark, M. Li, S. Riddle, and R. Bowen. 2020. Metabolism and inflammation predict cardiopulmonary disease outcomes in fattening beef cattle: histopathology. Conf. Res. Workers Anim. Disease. Virtual. Abstr. 284.


Progress 07/15/19 to 07/14/20

Outputs
Target Audience:Two target audiences have been reached by this research. 1. Scientists with focus on Animal Health, and 2. Beef cattleindustry professionals involved in management of feedlot cattle and their health as they progress to harvest. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Most importantly, salary of Greta Krafsur (DVM, Pathology Resident, and CSU PhD student)is not funded by this project; however, this project contributes to the PhD dissertation research of Dr. Krafsur. Greta was initially funded as a postdoc/research fellow by an NIH trainingprogram at the University of Colorado-Denver, Anshutz Medical School. However, Dr. Krafsur is now a pathologist for the State of South Dakota Veterinary Diagnostic Laboratory. After all of the accolades, what is important is that Dr. Krafsur is actively working and being trained by this project and we look forward to the histopathology analyses she is doing with cardiopulmonary tissues harvested from high and low PAP feedlot steers. How have the results been disseminated to communities of interest?In addition to an abstract presented to the animal health scientific communityat the100thConference ofResearchWorkers AnimalDisease in November2019, the following presentations were made to feedlot managers and veterinarians: Heart failure in beef cattle - PAP EPD development.International Genetic Solutions (IGS) booth seminar at National Cattlemen's Beef Association meeting, San Antonio, TX February 5, 2020. Mid-late feeding mortalities in finishing cattle and potential causes.Zoetis WebEx for Global Technical Services and Veterinary Medical Research and Development. November, 6, 2019. Late feeding mortality and pulmonary hypertension.Think-tank for "Late Day Morbidity Challenges in High Performance Cattle." Noble Foundation, Ardmore, OK, September 19, 2019. What do you plan to do during the next reporting period to accomplish the goals?Most importantly, we will complete laboratory/histopathology analyses as well as RNA-Seq analyses of gene expression that can potentially yield biomarkers indicative of pulmonary hypertension. Overall, we will focus on data analyses and writing of results.We expect to complete the project as scheduled; however, being out of office and laboratories because of COVID19 is slowing progress.

Impacts
What was accomplished under these goals? Animal work of this project was completed in spring of 2019. Subsequently, summer/fall of 2019 to present, efforts have focused on data analysis, preparation of tissue samples for histopathology, and RNA-sequencing of transcripts from peripheral blood mononucleocytes (PBMC-white blood cells/buffy coat). To date, parts of Specific Aim number 3a have been accomplished. As per abstract publication from December of 2019, the following results were presented: Angus steers were fed a growth-finishing ration targeting a gain of 1.5 kg/day for 9 months thru Winter and Spring months (northern hemisphere 2018-2019). Steers were initially managed at the Eastern Colorado Research Center, and those determined to be in the groupings of High and Low PAP (n = 45), were placed in a feed intake evaluation system. The PAP assessments averaged 44.7±1.3 (3 months), 52.3±1.9 (6 months), 65.1±4.0 (9 months) mmHg and the increase in PAP was attributed to increased variance in the High PAP group (i.e. maximal measurement at 3 versus 9 months of age was 66 vs 122 mmHg). The High and Low PAP groups prior to harvest averaged 87.2 and 44.1±5.3 mmHg, respectively (P < 0.01). Death loss was 5.5% with all of these mortalities attributed to PH-induced heart malformations and potential interaction with respiratory disease in the High PAP group. Eight-six percent of the mortalities occurred after 6 months of feeding. Body weight was similar among the two groups (583±7.3 kg); however, the Low PAP steers had better (P < 0.01) average daily gain (1.5 > 1.3±0.05 kg/d and feed efficiency (feed:gain in kg; 2.7 < 3.5±0.2)than the High PAP steers.The remaining contemporaries to these cattle had harvest traits of 37% prime and yield grade of 3.7±0.1. Also, 3b was accomplished as the infection with BRSV has little to no effect on these steers. To reiterate, obtaining RNA-seq data from PBMC of high vs low PAP steers is an important step forward for this study. The sequencing is completed; however, our analysis of these data has been slowed due to challenges of COVID19.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: M.G. Thomas, M.M. Culbertson, T. Holt, R.D. Brown, G.M. Krafsur, S.E. Speidel, R.M. Enns, R. Bowen, M. Li, and K. Stenmark. 2019. Metabolism and inflammation predict cardiopulmonary disease outcomes in fattening beef cattle.: Animal Model. 100th Conf. Res. Workers Anim. Disease. P165, Chicago, Ill. November 3-5, 2019.


Progress 07/15/18 to 07/14/19

Outputs
Target Audience:Animal Agriculture Scientific community and beef cattle feeding industries, which includes ruminant nutritionists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project is providing samples for the PhD project of Greta Krafsur (DVM). Dr. Krafsur has become a board certified pathologist and this research will help her complete her PhD. How have the results been disseminated to communities of interest?The beef cattle feeding industry has become very interested in this project, especially as its relates to feedlot mortality due to pulmonary hypertension. Therefore, invited speaking opportunities are below: Plains Nutrition Conference, San Antonio, TX, April 2019. Beef Improvement Federation, Brookings, South Dakota, June 2019 High Plains Beef Symposium, Beef Speices, American Socity of Animal Science Annual Meeting, Austin, TX, July 2019. What do you plan to do during the next reporting period to accomplish the goals?Laboratory and data analyses will begin; therefore, advancement to achieving goals of the project will be much more evident.

Impacts
What was accomplished under these goals? The animal handling, data and tissuecollection process for this projects has been completed. 107 Angus steers were fed for ~9 months and a 2 x 2 factorial project executed. The main effects were High versus Low pulmonary arterial pressure group (PAP, indicator trait for hypoxia and pulmonary hypertension in cattle)and control-vehicle (VHC) versus virsus challenge with bovine respiratory syncytial virus (BRSV).

Publications