Smoothing spline assessment of the accuracy of enteric hydrogen and methane production measurements from dairy cattle using various sampling schemes.

Estimating daily enteric hydrogen (H2) and methane (CH4) emitted from dairy cattle using spot sampling techniques requires accurate sampling schemes. These sampling schemes determine the number of daily samplings and their intervals. This simulation study assessed the accuracy of daily H2 and CH4 emissions from dairy cattle using various sampling schemes for gas collection. Gas emission data were available from a crossover experiment with 28 cows fed twice daily at 80% to 95% of the ad libitum intake, and an experiment that used a repeated randomized block design with 16 cows twice daily fed ad libitum. Gases were sampled every 12 to 15 min for 3 consecutive days in climate respiration chambers. Feed was fed in 2 equal portions per day in both experiments. Per individual cow-period combination, generalized additive models were fitted to all diurnal H2 and CH4 emission profiles. Per profile, the models were fitted using the generalized cross-validation, REML, REML while assuming correlated residuals, and REML while assuming heteroscedastic residuals. The areas under the curve (AUC) of these 4 fits were numerically integrated over 24 h to compute the daily production and compared with the mean of all data points, which was considered the reference. Next, the best of the 4 fits was used to evaluate 9 different sampling schemes. This evaluation determined the average predicted values sampled at 0.5, 1, and 2 h intervals starting at 0 h from morning feeding, at 1 and 2 h intervals starting at 0.5 h from morning feeding, at 6 and 8 h intervals starting at 2 h from morning feeding, and at 2 unequally spaced intervals with 2 or 3 samples per day. Sampling every 0.5 h was needed to obtain daily H2 productions not different from the selected AUC for the restricted feeding experiment, whereas less frequent sampling had predictions varying from 47% to 233% of the AUC. For the ad libitum feeding experiment, sampling schemes had H2 productions from 85% to 155% of the corresponding AUC. For the restricted feeding experiment, daily CH4 production needed samplings every 2 h or shorter, or 1 h or shorter, depending on sampling time after feeding, whereas sampling scheme did not affect CH4 production for the twice daily ad libitum feeding experiment. In conclusion, sampling scheme had a major impact on predicted daily H2 production, particularly with restricted feeding, whereas daily CH4 production was less severely affected by sampling scheme.

Evaluating growth response of broiler chickens fed diets supplemented with synthetic DL-methionine or DL-hydroxy methionine: a meta-analysis.

Methionine (Met) is the first limiting amino acid in corn and soybean meal-based diets (containing L-Met) in broiler chickens, which are often supplemented with synthetic DL-Met or DL-Hydroxy Met (OH-Met). Our objective was to quantitatively assess the efficacy of synthetic Met sources and determine differences in growth rate of broilers fed at or below requirements in response to Met intake. A systematic literature search resulted in building a database containing 480 treatment means from 39 articles published between 1985 and 2019 globally. The database was divided into starter, grower, and finisher subsets based on the age of the broilers. For each subset, linear-plateau and quadratic-plateau models were fitted to determine Met or sulfur amino acid (SAA; Met + Cysteine) requirements using average daily gain as a response variable. For each phase, 4 new subsets were obtained by only retaining records with digestible Met or SAA intake at or below requirement by linear-plateau or quadratic-plateau models. Then, a linear model (without plateau) was fitted for all new subsets for each rearing phase using supplemental digestible synthetic Met or SAA intake (basal Met intake was subtracted from total Met intake) as independent variables. The basal diet was made of only raw materials without supplementation of any synthetic Met source. Finally, the models were extended to evaluate source of synthetic Met effects on the slope parameter. At all stages of model fitting, the inclusion of a random study effect was evaluated for each parameter. All models were fitted within a Bayesian framework, for which minimally informative priors were used. The best models, that is, the most accurate inclusion of random effects, were selected based on at least 10-point difference in leave-one-out cross-validation information criterion. Model selection criteria did not consistently favor either of the linear- and quadratic-plateau models to determine Met or SAA requirements across broiler growth phases. Extending models with covariates (e.g., dietary energy and amino acids) did not improve any model fit. Body weight gain response of broiler chickens to the 2 sources was not different when fed at or below Met requirements for any of the growth phases.

Inhibited Methanogenesis in the Rumen of Cattle: Microbial Metabolism in Response to Supplemental 3-Nitrooxypropanol and Nitrate.

3-Nitrooxypropanol (3-NOP) supplementation to cattle diets mitigates enteric CH4 emissions and may also be economically beneficial at farm level. However, the wider rumen metabolic response to methanogenic inhibition by 3-NOP and the N O 2 - intermediary metabolite requires further exploration. Furthermore, N O 3 - supplementation potently decreases CH4 emissions from cattle. The reduction of N O 3 - utilizes H2 and yields N O 2 - , the latter of which may also inhibit rumen methanogens, although a different mode of action than for 3-NOP and its N O 2 - derivative was hypothesized. Our objective was to explore potential responses of the fermentative and methanogenic metabolism in the rumen to 3-NOP, N O 3 - and their metabolic derivatives using a dynamic mechanistic modeling approach. An extant mechanistic rumen fermentation model with state variables for carbohydrate substrates, bacteria and protozoa, gaseous and dissolved fermentation end products and methanogens was extended with a state variable of either 3-NOP or N O 3 - . Both new models were further extended with a N O 2 - state variable, with N O 2 - exerting methanogenic inhibition, although the modes of action of 3-NOP-derived and N O 3 - -derived N O 2 - are different. Feed composition and intake rate (twice daily feeding regime), and supplement inclusion were used as model inputs. Model parameters were estimated to experimental data collected from the literature. The extended 3-NOP and N O 3 - models both predicted a marked peak in H2 emission shortly after feeding, the magnitude of which increased with higher doses of supplement inclusion. The H2 emission rate appeared positively related to decreased acetate proportions and increased propionate and butyrate proportions. A decreased CH4 emission rate was associated with 3-NOP and N O 3 - supplementation. Omission of the N O 2 - state variable from the 3-NOP model did not change the overall dynamics of H2 and CH4 emission and other metabolites. However, omitting the N O 2 - state variable from the N O 3 - model did substantially change the dynamics of H2 and CH4 emissions indicated by a decrease in both H2 and CH4 emission after feeding. Simulations do not point to a strong relationship between methanogenic inhibition and the rate of N O 3 - and N O 2 - formation upon 3-NOP supplementation, whereas the metabolic response to N O 3 - supplementation may largely depend on methanogenic inhibition by N O 2 - .

Bayesian mechanistic modeling of thermodynamically controlled volatile fatty acid, hydrogen and methane production in the bovine rumen.

Dynamic modeling of mechanisms driving volatile fatty acid and hydrogen production in the rumen microbial ecosystem contributes to the heuristic prediction of CH4 emissions from dairy cattle into the environment. Existing mathematical rumen models, however, lack the representation of these mechanisms. A dynamic mechanistic model was developed that simulates the thermodynamic control of hydrogen partial pressure ( [Formula: see text] ) on volatile fatty acid (VFA) fermentation pathways via the NAD+ to NADH ratio in fermentative microbes, and methanogenesis in the bovine rumen. This model is unique and closely aligns with principles of reaction kinetics and thermodynamics. Model state variables represent ruminal carbohydrate substrates, bacteria and protozoa, methanogens, and gaseous and dissolved fermentation end products. The model was extended with static equations to model the hindgut metabolism. Feed composition and twice daily feeding were used as model inputs. Model parameters were estimated to experimental data using a Bayesian calibration procedure, after which the uncertainty of the parameter distribution on the model output was assessed. The model predicted a marked peak in [Formula: see text] after feeding that rapidly declined in time. This peak in [Formula: see text] caused a decrease in NAD+ to NADH ratio followed by an increased propionate molar proportion at the expense of acetate molar proportion, and an increase in CH4 production that steadily decreased in time, although the magnitude of increase for CH4 emission was less than for [Formula: see text] . A global sensitivity analysis indicated that parameters that determine the fractional passage rate and NADH oxidation rate altogether explained 86% of the variation in predicted daily CH4 emission. Model evaluation indicated over-prediction of in vivo CH4 emissions shortly after feeding, whereas under-prediction was indicated at later times. The present rumen fermentation modeling effort uniquely provides the integration of the [Formula: see text] controlled NAD+ to NADH ratio for dynamically predicting metabolic pathways that yield VFA, H2 and CH4.

Effect of Mootral-a garlic- and citrus-extract-based feed additive-on enteric methane emissions in feedlot cattle.

Enteric methane (CH4) production is the main source of greenhouse gas emissions from livestock globally with beef cattle contributing 5.95% of total global greenhouse gas emissions. Various mitigation strategies have been developed to reduce enteric emissions with limited success. In vitro studies have shown a reduction in CH4 emissions when using garlic and citrus extracts. However, there is paucity of data regarding in vivo studies investigating the effect of garlic and citrus extracts in cattle. The objective of this study was to quantitatively evaluate the response of Angus × Hereford cross steers consuming the feed additive Mootral, which contains extracts of both garlic and citrus, on CH4 yield (g/kg dry matter intake [DMI]). Twenty steers were randomly assigned to two treatments: control (no additive) and Mootral supplied at 15 g/d in a completely randomized design with a 2-wk covariate and a 12-wk data collection periods. Enteric CH4 emissions were measured using the GreenFeed system during the covariate period and experimental weeks 2, 6, 9, and 12. CH4 yield (g/kg DMI) by steers remained similar in both treatments for weeks 2 to 9. In week 12, there was a significant decrease in CH4 yield (23.2%) in treatment compared to control steers mainly because the steers were consuming all the pellets containing the additive. However, overall CH4 yield (g/kg DMI) during the entire experimental period was not significantly different. Carbon dioxide yield (g/kg DMI) and oxygen consumption (g/kg DMI) did not differ between treatments during the entire experimental period. DMI, average daily gain, and feed efficiency also remained similar in control and supplemented steers. The in vivo results showed that Mootral may have a potential to be used as a feed additive to reduce enteric CH4 production and yield in beef cattle but needs further investigation under various dietary regimen.

Diurnal Dynamics of Gaseous and Dissolved Metabolites and Microbiota Composition in the Bovine Rumen.

Diurnal patterns of ruminal fermentation metabolites and microbial communities are not commonly assessed when investigating variation in ruminal CH4 production. The aims of this study were to monitor diurnal patterns of: (i) gaseous and dissolved metabolite concentrations in the bovine rumen, (ii) H2 and CH4 emitted, and (iii) the rumen microbiota. Furthermore, the effect of dietary inclusion of linseed oil on these patterns was assessed. Four rumen cannulated multiparous cows were used in a cross-over design with two 17 days periods and two dietary treatments: a control diet and a linseed oil supplemented diet [40% maize silage, 30% grass silage, 30% concentrate on dry matter (DM) basis for both diets; fat contents of 33 vs. 56 g/kg of DM]. On day 11, rumen contents were sampled for 10 h after morning feeding to profile gaseous and dissolved metabolite concentrations and microbiota composition. H2 and CH4 emission (mass per unit of time) was measured in respiration chambers from day 13 to 17. A 100-fold increase in ruminal H2 partial pressure (contribution to the total pressure of rumen headspace gases) was observed at 0.5 h after feeding. This peak was followed by a decline to basal level. Qualitatively similar patterns after feeding were also observed for H2 and CH4 emission, ethanol and lactate concentrations, and propionate molar proportion, although the opposite pattern was seen for acetate molar proportion. Associated with these patterns, a temporal biphasic change in the microbial composition was observed as based on 16S ribosomal RNA with certain taxa specifically associated with each phase. Bacterial concentrations (log10 16S ribosomal RNA gene copies based) were affected by time, and were increased by linseed oil supplementation. Archaeal concentrations (log10 16S ribosomal RNA gene copies based) tended to be affected by time and were not affected by diet, despite linseed oil supplementation decreasing CH4 emission, tending to decrease the partial pressure of CH4, and tending to increase propionate molar proportion. Linseed oil supplementation affected microbiota composition, and was most associated with an uncultivated Bacteroidales taxon. In summary, our findings support the importance of diurnal dynamics for the understanding of VFA, H2, and CH4 production.

The Contribution of Mathematical Modeling to Understanding Dynamic Aspects of Rumen Metabolism.

All mechanistic rumen models cover the main drivers of variation in rumen function, which are feed intake, the differences between feedstuffs and feeds in their intrinsic rumen degradation characteristics, and fractional outflow rate of fluid and particulate matter. Dynamic modeling approaches are best suited to the prediction of more nuanced responses in rumen metabolism, and represent the dynamics of the interactions between substrates and micro-organisms and inter-microbial interactions. The concepts of dynamics are discussed for the case of rumen starch digestion as influenced by starch intake rate and frequency of feed intake, and for the case of fermentation of fiber in the large intestine. Adding representations of new functional classes of micro-organisms (i.e., with new characteristics from the perspective of whole rumen function) in rumen models only delivers new insights if complemented by the dynamics of their interactions with other functional classes. Rumen fermentation conditions have to be represented due to their profound impact on the dynamics of substrate degradation and microbial metabolism. Although the importance of rumen pH is generally acknowledged, more emphasis is needed on predicting its variation as well as variation in the processes that underlie rumen fluid dynamics. The rumen wall has an important role in adapting to rapid changes in the rumen environment, clearing of volatile fatty acids (VFA), and maintaining rumen pH within limits. Dynamics of rumen wall epithelia and their role in VFA absorption needs to be better represented in models that aim to predict rumen responses across nutritional or physiological states. For a detailed prediction of rumen N balance there is merit in a dynamic modeling approach compared to the static approaches adopted in current protein evaluation systems. Improvement is needed on previous attempts to predict rumen VFA profiles, and this should be pursued by introducing factors that relate more to microbial metabolism. For rumen model construction, data on rumen microbiomes are preferably coupled with knowledge consolidated in rumen models instead of relying on correlations with rather general aspects of treatment or animal. This helps to prevent the disregard of basic principles and underlying mechanisms of whole rumen function.

Correction: Thermodynamic Driving Force of Hydrogen on Rumen Microbial Metabolism: A Theoretical Investigation.

[This corrects the article DOI: 10.1371/journal.pone.0161362.].

Thermodynamic Driving Force of Hydrogen on Rumen Microbial Metabolism: A Theoretical Investigation.

Hydrogen is a key product of rumen fermentation and has been suggested to thermodynamically control the production of the various volatile fatty acids (VFA). Previous studies, however, have not accounted for the fact that only thermodynamic near-equilibrium conditions control the magnitude of reaction rate. Furthermore, the role of NAD, which is affected by hydrogen partial pressure (PH2), has often not been considered. The aim of this study was to quantify the control of PH2 on reaction rates of specific fermentation pathways, methanogenesis and NADH oxidation in rumen microbes. The control of PH2 was quantified using the thermodynamic potential factor (FT), which is a dimensionless factor that corrects a predicted kinetic reaction rate for the thermodynamic control exerted. Unity FT was calculated for all glucose fermentation pathways considered, indicating no inhibition of PH2 on the production of a specific type of VFA (e.g., acetate, propionate and butyrate) in the rumen. For NADH oxidation without ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity to zero for different NAD+ to NADH ratios and pH of 6.2 and 7.0, which indicates thermodynamic control of PH2. For NADH oxidation with ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity at pH of 7.0 only. For the acetate to propionate conversion, FT increased from 0.65 to unity with increasing PH2, which indicates thermodynamic control. For propionate to acetate and butyrate to acetate conversions, FT decreased to zero below the rumen range of PH2, indicating full thermodynamic suppression. For methanogenesis by archaea without cytochromes, FT differed from unity only below the rumen range of PH2, indicating no thermodynamic control. This theoretical investigation shows that thermodynamic control of PH2 on individual VFA produced and associated yield of hydrogen and methane cannot be explained without considering NADH oxidation.