Source: UNIVERSITY OF CALIFORNIA, DAVIS submitted to NRP
EVALUATION OF COMPREHENSIVE MECHANISTIC MODEL OF ENTERIC FERMENTATION IN DAIRY CATTLE UNDER METHANE-INHIBITING FEED ADDITIVE SUPPLEMENTATION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1032602
Grant No.
2024-67011-43009
Cumulative Award Amt.
$60,000.00
Proposal No.
2023-11578
Multistate No.
(N/A)
Project Start Date
Aug 15, 2024
Project End Date
Aug 14, 2025
Grant Year
2024
Program Code
[A7101]- AFRI Predoctoral Fellowships
Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671
Performing Department
(N/A)
Non Technical Summary
To feed the growing human population, global food production needs to increase. However, this agricultural intensification must be sustainable and protect both food security and environmental health, as agriculture is a major driver of climate change through enteric fermentation. Enteric fermentation is the process by which feed is broken down by microbes in the rumen (specialized stomach compartment of cows), which allows cattle to utilize fibrous plants as energy sources, unlike many other animals. However, enteric fermentation also results in the production of methane, a powerful greenhouse gas. Recent advances in our understanding of how this methane is produced have led to the development of substances added to an animal's feed, or Animal Feed Additives (AFA), that can reduce enteric methane emissions. However, many interacting factors impact methane production, such as cattle type, weight, feed intake, and the composition of the cow's diet. In addition, AFA can have indirect effects that may increase the excretion of other environmental pollutants by cows. To reduce methane as much as possible with AFA, we need to know how the effectiveness of AFA depends on all these interacting factors. However, this can be very challenging to assess in experiments with real animals, where it is difficult or impossible to measure all the possible interacting variables. To complement studies in real animals, animal scientists also use mathematical models, or a series of equations that describe how the cow's feed is turned into methane, to answer questions about enteric fermentation and how it may be affected by AFA.I have previously developed a comprehensive mathematical model of enteric fermentation in dairy cows that is designed to answer these questions. My model predicts how much methane a cow will produce, as well as many other variables, depending on what she is eating and which and how much AFA she is fed. However, I have not yet assessed how accurate the model's predictions are, what parts of the model are most important to making its predictions better, or how certain or uncertain the model's predictions are. In this project, my objectives are to: 1) evaluate the accuracy of the model's predictions in several different scenarios and to compare this accuracy to that of similar models; 2) conduct analyses that will tell me which parts of the model are the most important to its predictions, and 3) assess how confident we are in the model's predictions, given uncertainties in inputs to the model. After I evaluate the model with these analyses, my last objective is to use the model to find a diet and AFA combination that most effectively decreases methane emissions from dairy cows.Achieving these goals will confirm that the model is a useful, accurate tool that animal scientists and policy makers feel confident using to answer questions about the interactions between AFA use and methane production. We can then use this tool to find ways to maximize AFA efficiency, minimize greenhouse gas emissions from dairy cows, or ask more basic questions about enteric fermentation, ultimately helping us reduce the environmental impacts of animal agriculture.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3023499101033%
3027299101067%
Goals / Objectives
The overall goal of this project is to evaluate a previously developed mechanistic model of rumen fermentation and use this model to optimize methane-inhibiting animal feed additive (AFA) use. This goal is divided into three objectives:Evaluate model predictive performance on independent datasetsSub aim 1a: Assess model predictive performance.Sub aim 1b: Compare model performance to similar models.Evaluate sensitivity of model to fitted parameter values, calibrate parameters, and quantify uncertainty in model outputsSub aim 2a: Conduct global sensitivity analysis.Sub aim 2b: Calibrate parameters.Sub aim 2c: Conduct uncertainty analysis.Optimize basal diet and AFA implementation scheme for maximum efficacy.
Project Methods
Aim 1: Evaluate model predictive performance on independent dataset and compare performance to related models. Sub aim 1a: Assess model predictive performance. An evaluation dataset of hourly fermentation dynamics from dairy cattle will be used to assess model predictions of diurnal enteric CH4 and H2 emissions, VFA concentrations, microbial dynamics, and rumen pH from simulations with no AFA supplementation. Data from an A. taxiformis trial in dairy cattle will be used to assess performance of daily enteric CH4 predictions from simulations including bromoform supplementation. A dataset of dairy cattle supplemented with 3NOP will include model performance as assessed by mean square prediction error (MSPE) and square root of MSPE (RMSPE) of the model's predictions from observations in each test dataset. MPSE will be decomposed into mean, slope, and random bias. The ratio of the RMSPE to the standard deviation of the observations (RSR) will also be calculated and used to compare performance across datasets.Sub aim 1b: Compare model performance to similar models. Model performance on the 3NOP dataset will be assessed for a rumen mechanistic model incorporating 3NOP using the same performance metrics as above, while model performance on the A. taxiformis dataset will be assess for a mechanistic model incorporating A. taxiformis. Evaluation milestones for Aim 1b will include error index statistics (MSPE, RMSPE, and RSR) for each model on each test dataset. To assess relative model performance, Lin's Concordance Correlation Coefficient (CCC) will calculated and decomposed into the Pearson correlation coefficient, r, which measures prediction precision, and the bias correction factor, Cb, which measures prediction accuracy.Aim 2: Evaluate sensitivity of model to fitted parameter values, calibrate parameters, and quantify uncertainty in model outputs.Sub aim 2a: Conduct global sensitivity analysis. Local sensitivity analysis (SA) explores the sensitivity of model outputs to parameters by holding all parameters except one constant, and therefore does not account for parameter correlation. Global SA (GSA) overcomes this limitation by varying all inputs at the same time, and measuring the variance attributed to an input relative to the total variance of all inputs. However, regression-based GSA approaches are only applicable if the model is linear in its parameters, which may not be known a priori. Therefore, a model-free, global, variance-based SA approach will be used. Because calculating variance-based sensitivity indices for all model parameters is computationally expensive, we will first screen all parameters using the Method of Morris. The Morris method provides the measures mu*, the mean of the elementary effects of a given parameter, and sigma*, the standard deviation of the elementary effects of the parameter. We will select all parameters whose normalized (mu*, sigma*) coordinate pair is a distance of 0.5 or greater from the origin for further analysis. Parameters not selected for further analysis will be fixed to their optimized values. We will conduct a GSA by calculating the first- and total-order Sobol' indices for each selected parameter. Sobol' indices decompose variance in the model output into first-order and higher order effects of each parameter, respectively. These indices will quantify the contribution of the variance in each parameter to the variance in model outputs due to their first and higher order effects.Sub aim 2b: Calibrate parameters. The previously accomplished parameter optimization deterministically identified parameters values that minimize model errors on fitting data, but did not estimate parameter distributions or uncertainty. We will utilize a Markov Chain Monte Carlo simulation to sample values from each parameter posterior distribution that can be used to estimate mean, variance, and 95% credible intervals for all influential parameters.Sub aim 2c: Model output uncertainty analysis. We will perform an uncertainty analysis of model outputs by executing the model many times, sampling parameters from their joint posterior distribution as generated by the MCMC simulation, and the mean and standard deviation of model outputs will be estimated, quantifying the variability in all outputs due to variability in the parameters.Aim 3: Optimize basal diet and AFA implementation scheme for maximizing AFA efficacy. An optimization algorithm will be used to optimize basal diet and AFA choices that respectively minimize enteric CH4, net GHG emissions, or nutrient excretion to the environment as predicted by the mechanistic model, given fixed AFA doses. Outcomes will include a description of optimal basal diet for each AFA and their associated environmental impacts.Interpretation of results from Aims 1-3: Aim 1 will assess the model's ability to make accurate predictions and compare its accuracy to similar models. Aim 2a will identify the most globally influential model influential parameters, which I expect to include pH- and fractional outflow-related parameters. Results of Aims 1 and 2a can be used together to identify parameters with disproportionate effects on model outputs that may cause poor model predictions. Given these results, I will reconsider model structure, particularly novel elements of outflow and pH, to further improve model performance. Aims 2b and 2c will generate likely ranges for each model prediction variable based on uncertainty in inputs, which will allow us to understand the typical variation in each outcome variable and move away from deterministic "average animal" predictions. In Aim 3, we will identify diet and AFA protocols that minimize various environmental impacts, and together, Aims 2b-c and 3 will allow us to understand the possible range of effects these diets may have.Evaluation of expected outcomes: The expected outcomes of Aim 1 will provide quantitative information on the predictive performance of the mechanistic model and therefore evaluate the hypothesis embedded in model, such as the importance of pH, rumen outflow, and thermodynamics in determining enteric CH4 emissions. Evaluation of these hypotheses is an important step in developing our understanding of the effects of AFA on enteric fermentation and consolidating various hypotheses in the scientific literature on enteric fermentation into one comprehensive rumen model. Outcomes from Aim 1 will also allow fellow researchers to potentially use the evaluated model to investigate feed additive use and related questions, adding a complementary tool to in vivo studies. Results from Aim 2 will allow improved prediction of on-farm cattle GHG emissions with associated uncertainties, improving our understanding of likely ranges of these outcomes. Results from Aim 3 will identify how to implement methane-inhibiting AFA most effectively and allow the potential use of identified optimum diets to maximize AFA efficacy on-farm. The evaluation of all three aims, together, will lead to the potential reduction of agricultural GHG emissions through optimized use of methane-inhibiting feed additives.

Progress 08/15/24 to 08/14/25

Outputs
Target Audience:Animal scientists working in ruminant nutrition and microbiology to understand and mitigate enteric methane emissions; ruminant nutritionists working to optimize methane-inhibiting feed additive use in rations; mathematical biologists interested in theoretical aspects of the development and evaluation of mechanistic models of biological systems Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project supported the development of the graduate student Project Director into an independent researcher by providing financial support for their dissertation year, culminating in the successful submission of the doctoral dissertation and completion of doctoral degree. In addition, funding from the project provided full support for the Project Director to attend the 2025 annual meeting of the American Dairy Science Association and companion symposium Meeting of the Animal Science Modeler's Group, where the work resulting from the project was presented and opportunities for professional development for graduate students and networking were provided. How have the results been disseminated to communities of interest?The target audience of animal scientists working in ruminant nutrition and microbiology to understand and mitigate enteric methane emissions were reached through oral presentations by the Project Director (PD) at the American Dairy Science Association's Annual Meeting and companion meeting of the Animal Science Modeler's Group (June 2025, Louisville, Kentucky). The PD presented results from the evaluation and analysis of the mechanistic model of fermentation that was the subject of this project, discussing variables at the animal and rumen level that impact methane emissions. The target audience of ruminant nutritionists working to optimize methane-inhibiting feed additive use in rations were reached through the same oral presentations presenting the results from completed work of the application of the mechanistic model of rumen fermentation to optimize diets for maximum methane-inhibiting feed additive use efficacy, and will be reached through the publication of the PD's PhD dissertation reporting these results. Mathematical biologists interested in theoretical aspects of the development and evaluation of mechanistic models of biological systems were reached through publication of a review article on mathematical approaches used in mechanistic models of rumen fermentation, and will be reached through the publication of the PD's PhD dissertation reporting the methods used to evaluate and perform sensitivity analysis of the mathematical model. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The overall goal of this project was to evaluate a previously developed mechanistic model of rumen fermentation and use this model to optimize antimethanogenic feed additive (AMFA) use. This goal was divided into three objectives, with sub aims and accomplishments related to each sub aim noted below. Sub aims 1a-b: Evaluate model predictive performance on independent datasets and compare model performance to similar models. Accomplishments: Model predictions of diurnal CH4, CO2, and H2 production and other rumen parameters were evaluated against independent in vivo data under different AMFA supplementation scenarios. Model predictions were compared to treatment group averages at 17 diurnal timepoints from two separate studies: one with dairy cattle receiving no AMFA supplementation and another with dairy cattle supplemented with bromoform (CHBr3)-containing seaweed at 0.5 and 1% organic matter (OM). Diurnal CH4 predictions were also evaluated against data from dairy cattle supplemented with 3NOP at 6 doses. In addition, average daily CH4, H2, and CO2 production, rumen fluid VFA and ammonia concentrations, pH, and microbial population variables were assessed across up to 13 treatment groups in studies with cattle under 3NOP supplementation. The model demonstrated the lowest prediction error for diurnal CH4 emissions under the 3NOP scenarios. Prediction error for diurnal CH4 was similar under the base scenario compared to the low CHBr3 scenario but greater under high CHBr3 scenario. Predictions of diurnal CH4 were more accurate than CO2 and H2 predictions under the base scenario, while predictions of average daily H2 under the 3NOP scenario outperformed CO2 predictions. Diurnal methane predictions under low and high CHBr3 scenarios were more accurate than CO2 and H2 predictions. The model's predictions were directly compared to a similar model of enteric fermentation under 3NOP supplementation and the model's predicted changes in enteric gas emissions due to bromoform were compared to a similar in vitro model of rumen fermentation. RMSPE for CH4 emissions was similar to these models. Our results provide quantitative information on the predictive performance of the mechanistic model and, along with subsequent sensitivity analysis, evaluate the hypotheses embedded in the model, such as the representations of pH, rumen outflow, and thermodynamics. This evaluation is an important step toward consolidating findings on the effects of AMFA on enteric fermentation into a comprehensive rumen model. Sub aims 2a-c: Evaluate sensitivity of model to fitted parameter values, calibrate parameters, and quantify uncertainty in model outputs. Accomplishments: The Morris screening method of sensitivity analysis identified parameters related to methanogenic H2 metabolism, carbohydrate and protein fermentation, protozoal protein metabolism and microbial predation as most influential for CH4 production. Under the CHBr3 scenario, fewer strongly influential parameters were identified. Parameters related to carbohydrate and protein fermentation, H2 emission and utilization, protozoal protein metabolism and microbial predation, and fibrolytic bacterial growth were most influential on the H2 emission rate across all AMFA scenarios. The most influential parameter by first-order Sobol' sensitivity index on the CH4 and H2 emission rate under the Base scenario, on the CH4 emission rate under the CHBr3 scenario, and on the H2 emission rate under the 3NOP scenario was the maximum velocity of H2 uptake by methanogens for CH4 formation. According to the UA, the most uncertain model outcome under the Base scenario was the fibrolytic bacterial storage polysaccharide pool. Comparisons of influential parameters, uncertain outcomes, and treatment effects across AMFA scenarios demonstrate how rumen mechanistic models can be used to investigate the impacts of inhibiting methanogenesis on the rumen environment. Aim 3. Optimize basal diet and AMFA implementation scheme for maximum efficacy. Accomplishments: Optimization was conducted for CH4 reduction efficacy using fixed doses of 3NOP and CHBr3. We also optimized dietary composition and additive dose to jointly minimize absolute enteric CH4 and GHG emissions and total manure N excretion. For the CHBr3-optimized diet, relative to the original diet, NDF decreased by 0.33%, starch decreased by 21%, and DMI increased by 8%, leading to a reduction in CH4 emissions of 8.8% from the original diet with CHBr3. For the 3NOP-optimized diet, starch increased by 5%, NDF decreased by 6%, and DMI decreased by 8%, leading to an 8.7% CH4 reduction. Diets optimized for minimizing GHG emissions and manure N excretion favored CHBr3 over 3NOP, possibly due to its metabolism to other nitrogenous compounds in the rumen. These results demonstrate how a complex mechanistic model of rumen fermentation can be applied to optimize the implementation of AMFA to maximize efficacy or minimize environmental impacts.

Publications

  • Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Pressman, E.M. and Kebreab, E., 2024. A review of key microbial and nutritional elements for mechanistic modeling of rumen fermentation in cattle under methane-inhibition. Frontiers in Microbiology, 15, p.1488370.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2025 Citation: E.M. Pressman and E. Kebreab, June 23, 2025. A mechanistic model predicts diurnal methane emissions in dairy cattle under methane mitigation. Oral presentation, 2025 American Dairy Science Association Annual Meeting.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2025 Citation: E.M. Pressman, 2025. Development and Evaluation of a Comprehensive Mechanistic Model of Enteric Fermentation in Dairy Cattle Supplemented with Methane-Inhibiting Feed Additives. PhD Dissertation, University of California, Davis.