Invited review: Large-scale indirect measurements for enteric methane emissions in dairy cattle: A review of proxies and their potential for use in management and breeding decisions.

Efforts to reduce the carbon footprint of milk production through selection and management of low-emitting cows require accurate and large-scale measurements of methane (CH4) emissions from individual cows. Several techniques have been developed to measure CH4 in a research setting but most are not suitable for large-scale recording on farm. Several groups have explored proxies (i.e., indicators or indirect traits) for CH4; ideally these should be accurate, inexpensive, and amenable to being recorded individually on a large scale. This review (1) systematically describes the biological basis of current potential CH4 proxies for dairy cattle; (2) assesses the accuracy and predictive power of single proxies and determines the added value of combining proxies; (3) provides a critical evaluation of the relative merit of the main proxies in terms of their simplicity, cost, accuracy, invasiveness, and throughput; and (4) discusses their suitability as selection traits. The proxies range from simple and low-cost measurements such as body weight and high-throughput milk mid-infrared spectroscopy (MIR) to more challenging measures such as rumen morphology, rumen metabolites, or microbiome profiling. Proxies based on rumen samples are generally poor to moderately accurate predictors of CH4, and are costly and difficult to measure routinely on-farm. Proxies related to body weight or milk yield and composition, on the other hand, are relatively simple, inexpensive, and high throughput, and are easier to implement in practice. In particular, milk MIR, along with covariates such as lactation stage, are a promising option for prediction of CH4 emission in dairy cows. No single proxy was found to accurately predict CH4, and combinations of 2 or more proxies are likely to be a better solution. Combining proxies can increase the accuracy of predictions by 15 to 35%, mainly because different proxies describe independent sources of variation in CH4 and one proxy can correct for shortcomings in the other(s). The most important applications of CH4 proxies are in dairy cattle management and breeding for lower environmental impact. When breeding for traits of lower environmental impact, single or multiple proxies can be used as indirect criteria for the breeding objective, but care should be taken to avoid unfavorable correlated responses. Finally, although combinations of proxies appear to provide the most accurate estimates of CH4, the greatest limitation today is the lack of robustness in their general applicability. Future efforts should therefore be directed toward developing combinations of proxies that are robust and applicable across diverse production systems and environments.

Plasma nitrogen isotopic fractionation and feed efficiency in growing beef heifers.

Fractionation of N isotopes occurs in many metabolic reactions which causes tissue proteins to become enriched in ¹5N while urea (urine) is depleted in ¹5N relative to the diet. We investigated ¹5N enrichment of whole plasma and its relationship with feed conversion efficiency (FCE) in growing beef heifers (n 84) offered 2 kg/d of concentrates with grass silage ad libitum. Heifers were on average 299 (SD 48·3) d old and weighed 311 (SD 48·8) kg. Plasma was obtained on day 79 (n 84) of the experiment and from a subset of animals (n 20) on four occasions between days 16 and 79. Silage DM intake (DMI) averaged 4·1 (SD 0·74) kg/d and concentrate DMI was 1·72 kg/d. Mean mid-test live weight was 333 (SD 47·6) kg, daily gain was 0·53 (SD 0·183) kg, FCE (g live-weight gain/g DMI) was 0·09 (SD 0·028) and residual feed intake (RFI) was 0 (SD 0·428). N isotopic fractionation (Δ¹5N; plasma δ¹5N - diet δ¹5N) averaged 3·58 ‰ on day 79 (n 84) and 3·90 ‰ for the subset of heifers. There was no relationship between Δ¹5N and RFI. Plasma δ¹5N and Δ¹5N were related to both FCE (negative) and animal weight (positive) for the whole population, and repeatable for the subset of animals over four time points. These relationships of Δ¹5N with FCE and animal weight are consistent with the anticipated negative relationship with N-use efficiency. There is potential to use Δ¹5N to provide rapid, low-cost estimates of FCE in cattle.

Short communication: Relationship between the efficiency of utilization of feed nitrogen and 15N enrichment in casein from lactating dairy cows.

The objective of the present study was to evaluate the relationship between the efficiency of conversion of feed N into milk N [N-use efficiency (NUE)] and (15)N enrichment of milk casein from lactating cows fed corn silage-based diets. Samples of feeds and milk were obtained from 3 experiments with lactating dairy cows. All diets were based on corn silage and were designed to evaluate the effects of (1) diets with different ratios of effective rumen-degradable protein to fermentable metabolizable energy (experiment 1), (2) different proportions of quickly and slowly rumen-degradable protein (experiment 2), and (3) synchronizing the availability of fermentable metabolizable energy and effective rumen-degradable protein in the rumen (experiment 3). Although no significant effect of diet on casein δ(15)N values was detected, casein was more enriched than the diet in each of the experiments. Nitrogen-use efficiency was negatively related to adjusted Δ(15)N (casein δ(15)N--diet δ(15)N) for experiments 1 and 2 individually and when combining data from all 3 experiments. The relatively low values for Δ(15)N suggest that these productive animals were using dietary N efficiently, with a high proportion of N going to milk protein and less to urea. The weak, although significant, relationship between NUE and adjusted Δ(15)N, is consistent with relatively little variation in hepatic deamination and transamination, with variation in rumen efficiency having the predominant effect on NUE. The present study confirms the lower (15)N enrichment in protein when NUE is high and the potential to use N-isotope fractionation as a marker of NUE.

Assessment of archaeol as a molecular proxy for methane production in cattle.

The objective of this study was to assess archaeol, a membrane lipid present in methanogenic Archaea, in cattle feces as a molecular proxy for methanogenesis in the rumen. Feces from 16 heifers either in early lactation [71 d in milk (DIM)] or mid lactation (120 DIM), consuming a diet consisting of 30/70 grass silage/concentrates [dry matter (DM) basis], were analyzed for archaeol. To prepare the feces for analysis, total lipids were obtained by Bligh-Dyer extraction, polar head groups were removed by acid methanolysis, an alcohol fraction was obtained by column chromatography, and finally, the alcohol fraction was trimethylsilylated before analysis by gas chromatography-mass spectrometry. Archaeol was quantified by comparison to an internal standard. A highly significant positive relationship was found between fecal archaeol concentration (mg/kg of DM) and methane output (g/kg of DM intake). A highly significant effect of stage of lactation on this relationship was observed. The significant relationship was surprising, given the lack of agreement between methane and total methanogens in previous studies using molecular biology techniques. Variation in the relationship between fecal archaeol concentrations and methane output could be attributed to differences in the methane-producing capability per cell and the selective retention of methanogens in the rumen. The effect of stage of lactation may have been due to differences in DM intake, affecting rumen passage rates and, thus, methanogen populations and activities.

The effect of strategic supplementation with trans-10,cis-12 conjugated linoleic acid on the milk production, estrous cycle characteristics, and reproductive performance of lactating dairy cattle.

The objective was to determine the effects of a protected (lipid-encapsulated) conjugated linoleic acid (LE-CLA) supplement on milk production, estrous cycle characteristics, and reproductive performance in lactating dairy cows on a pasture-based diet. Spring calving dairy cows (n=409) on a single pasture-based commercial dairy farm were used in a completely randomized block design. Cows were assigned to 1 of 2 dietary supplements [LE-CLA (n=203) or no supplement (control, n=206)]. The LE-CLA cows received 51 g/d of a lipid supplement containing 5 g of both trans-10,cis-12 and cis-9,trans-11 CLA from 0 to 60 d in milk. Milk samples were collected 3 times weekly, and each sample was analyzed for progesterone to determine the interval to first ovulation and estrous cycle characteristics. Milk yield and concentrations of fat, protein, and lactose were measured every 2 wk. Cows were inseminated following visual observation of estrus. The breeding season commenced on April 8, 2009 and continued for 16 wk. Transrectal ultrasonography was carried out at 30 to 36 d and 60 to 66 d post-AI to diagnose pregnancy. The LE-CLA treatment resulted in a decrease in milk fat concentration (36.9±0.06 g/kg vs. 30.7±0.06 g/kg for control and LE-CLA, respectively) and yield (0.91±0.02 kg/d vs. 0.84±0.02 kg/d for control and LE-CLA, respectively); however, milk yield was increased by LE-CLA supplementation (24.7±0.7 kg/d vs. 27.2±0.7 kg/d for control and LE-CLA, respectively), resulting in no overall difference in milk energy output. No effect of LE-CLA was observed on any estrous cycle characteristics or measures of reproductive performance. These results support that in pasture-based systems of dairy production, where energy intake limits milk production, energy spared by CLA-induced milk fat depression is partitioned toward increasing milk yield rather than toward body reserves.

Methane emissions and rumen metabolite concentrations in cattle fed two different silages.

In this study, 18 animals were fed two forage-based diets: red clover (RC) and grass silage (GS), in a crossover-design experiment in which methane (CH4) emissions were recorded in respiration chambers. Rumen samples obtained through naso-gastric sampling tubes were analysed by NMR. Methane yield (g/kg DM) was significantly lower from animals fed RC (17.8 ± 3.17) compared to GS (21.2 ± 4.61) p = 0.008. In total 42 metabolites were identified, 6 showing significant differences between diets (acetate, propionate, butyrate, valerate, 3-phenylopropionate, and 2-hydroxyvalerate). Partial least squares discriminant analysis (PLS-DA) was used to assess which metabolites were more important to distinguish between diets and partial least squares (PLS) regressions were used to assess which metabolites were more strongly associated with the variation in CH4 emissions. Acetate, butyrate and propionate along with dimethylamine were important for the distinction between diets according to the PLS-DA results. PLS regression revealed that diet and dry matter intake are key factors to explain CH4 variation when included in the model. Additionally, PLS was conducted within diet, revealing that the association between metabolites and CH4 emissions can be conditioned by diet. These results provide new insights into the methylotrophic methanogenic pathway, confirming that metabolite profiles change according to diet composition, with consequences for CH4 emissions.

Nitrogen partitioning and isotopic fractionation in dairy cows consuming diets based on a range of contrasting forages.

Nine multiparous Holstein-Friesian cows (initially 97 d in milk), were used in a 3×3 lattice square design experiment with 4-wk periods. All cows received 4 kg/d concentrates and dietary treatments were based on silages offered ad libitum: perennial ryegrass (PRG); timothy (TIM); tall fescue (TF); red clover (RC); red clover/corn silage mixture [40/60 on a dry matter (DM) basis; RCC]; red clover/whole-crop oat silage mixture (40/60 on a DM basis; RCO); or red clover/whole-crop oat silage mixture (25/75 on a DM basis; ORC). The remaining treatments were based on RCO with feed intake restricted to the level of PRG (RCOr) or with a low protein concentrate (50/50 mixture of barley and molassed sugar beet pulp; RCOlp). Experiment objectives were to evaluate diet effects on N partitioning and N isotopic fractionation. Yields of milk and milk protein were consistently high for diets RC, RCC, and RCO and low for the diets based on poorly ensiled grass silages. Restriction of intake (RCOr) and inclusion of a higher proportion of whole-crop oat silage (ORC) and the low-protein concentrate (RCOlp) led to some loss of production. Diet had little effect on milk fat, protein, and lactose concentrations: low concentrations of milk protein and lactose reflect the restricted energy intakes for all treatments. The highest diet digestibilities were measured for RC and PRG, whereas increasing inclusion of the whole-crop oat silage (0, 60, and 75% of forage DM) led to a marked decrease in diet digestibility (0.717, 0.624, and 0.574 g/g, respectively). Urinary excretion of purine derivatives, an indicator for rumen microbial protein synthesis, was significantly higher for RCC than for TIM and TF. Nitrogen intake ranged between 359 and 626 g/d (treatment means). Partitioning of N intake to feces and urine was closely related to N intake, although urinary N losses were less than predicted from N intake for the 60/40 mixtures of cereal silage and red clover silage. The 15N content of milk, urine, and feces were all influenced by diet 15N content. Isotopic fractionation meant that feces and milk were enriched and urine was depleted in 15N relative to the diet. Significant relationships were observed between the extent of enrichment of urine, feces, and milk, suggesting some commonality in fractionation pathways. The trend for the lowest 15N enrichment in milk protein occurring in diets with low N-use efficiency (milk N/feed N) was contrary to expectations, possibly because of endogenous contributions to milk protein or fractionation when dietary ammonia was incorporated into microbial protein.

Effects of dietary protein concentration and balance of absorbable amino acids on productive responses of dairy cows fed corn silage-based diets.

A cyclical changeover design experiment (3-wk periods; 12-wk total) was conducted to evaluate whether improving the balance of absorbable AA would allow the feeding of less crude protein (CP) without compromising production, thereby reducing the potential environmental pollution from dairy farms. Sixteen multiparous Holstein cows were assigned to 1 of 8 dietary treatments as total mixed rations (TMR) containing [dry matter (DM) basis] 45% corn silage, 5% coarsely chopped wheat straw, and 50% concentrate mixture. The 8 treatments were formulated to differ in dietary CP (14 and 16%; DM basis) and in the balance of absorbable AA achieved by changing the main protein source (MPS) of the concentrate mixtures [replacing soybean meal (SBM) with corn byproducts (CBP), dried corn distillers grains (DDG), and some corn gluten meal], and by adding a mixture of rumen-protected Lys and Met (RPLM). Feeding lactating dairy cows corn silage-based diets with 16% CP promoted significantly higher DM intakes and milk yields, and lower feed N-use efficiency than feeding diets with 14% CP. Replacing SBM with CBP significantly increased milk yields and decreased milk fat and protein concentrations, but had no effect on the efficiency of conversion of feed N into milk N. With 16% CP diets, the addition of RPLM decreased feed N use efficiency. A significant effect was observed for the MPS × RPLM interaction on milk protein concentrations. Plasma Lys concentration was lower with diets based on CBP, and plasma Met increased with RPLM. We did not find clear benefits of RPLM in facilitating a reduction of dietary protein without loss of production.

The effect of sheep genetic merit and feed allowance on nitrogen partitioning and isotopic discrimination.

Animal nitrogen (N) partitioning is a key parameter for profitability and sustainability of ruminant production systems, which may be predicted from N isotopic discrimination or fractionation (Δ15N). Both animal genetics and feeding level may interact and impact on N partitioning. Therefore, this study aimed to assess the interactive effects of genetic merit (G) and feed allowance (F) on N partitioning and Δ15N in sheep. The sheep were drawn from two levels of G (high G vs. low G; based on New Zealand Sheep Improvement Limited (http://www.sil.co.nz/) dual (wool and meat) growth index) and allocated to two levels of F (1.7 (high F) vs. 1.1 (low F) times Metabolisable Energy requirement for maintenance) treatments. Twenty-four Coopworth rams were divided into four equal groups for a N balance study: high G × high F, high G × low F, low G × high F, and low G × low F. The main factors (G and F) and the interaction term were used for 2-way ANOVA and regression analysis. Higher F led to higher N excretions (urinary N (UN); faecal N (FN); manure N), retained N, N use efficiency (NUE), and urinary purine derivatives excretion (P < 0.05). On the other hand, higher UN/N intake, and plasma Δ15N were observed with the lower F (P < 0.05). Higher G led to increased UN, FN, manure N, apparent N digestibility, and urinary purine derivatives excretion (P < 0.05). Higher F only increased UN in high G sheep, with no effect on low G sheep (P < 0.05). Regression analysis results demonstrated potential to use plasma Δ15N to reflect the effects of G and F on NUE and UN/N intake. Further research is urged to study interactive effects of genetic and feeding level on sheep N partitioning.

Review: Markers and proxies to monitor ruminal function and feed efficiency in young ruminants.

Developing the rumen's capacity to utilise recalcitrant and low-value feed resources is important for ruminant production systems. Early-life nutrition and management practices have been shown to influence development of the rumen in young animals with long-term consequences on their performance. Therefore, there has been increasing interest to understand ruminal development and function in young ruminants to improve feed efficiency, health, welfare, and performance of both young and adult ruminants. However, due to the small size, rapid morphological changes and low initial microbial populations of the rumen, it is difficult to study ruminal function in young ruminants without major invasive approaches or slaughter studies. In this review, we discuss the usefulness of a range of proxies and markers to monitor ruminal function and nitrogen use efficiency (a major part of feed efficiency) in young ruminants. Breath sulphide and methane emissions showed the greatest potential as simple markers of a developing microbiota in young ruminants. However, there is only limited evidence for robust indicators of feed efficiency at this stage. The use of nitrogen isotopic discrimination based on plasma samples appeared to be the most promising proxy for feed efficiency in young ruminants. More research is needed to explore and refine potential proxies and markers to indicate ruminal function and feed efficiency in young ruminants, particularly for neonatal ruminants.

Taxonomic annotation of 16S rRNA sequences of pig intestinal samples using MG-RAST and QIIME2 generated different microbiota compositions.

Environmental microbiome studies rely on fast and accurate bioinformatics tools to characterize the taxonomic composition of samples based on the 16S rRNA gene. MetaGenome Rapid Annotation using Subsystem Technology (MG-RAST) and Quantitative Insights Into Microbial Ecology 2 (QIIME2) are two of the most popular tools available to perform this task. Their underlying algorithms differ in many aspects, and therefore the comparison of the pipelines provides insights into their best use and interpretation of the outcomes. Both of these bioinformatics tools are based on several specialized algorithms pipelined together, but whereas MG-RAST is a user-friendly webserver that clusters rRNA sequences based on their similarity to create Operational Taxonomic Units (OTU), QIIME2 employs DADA2 in the construction of Amplicon Sequence Variants (ASV) by applying an error model that considers the abundance of each sequence and its similarity to other sequences. Taxonomic compositions obtained from the analyses of amplicon sequences of DNA from swine intestinal gut and faecal microbiota samples using MG-RAST and QIIME2 were compared at domain-, phylum-, family- and genus-levels in terms of richness, relative abundance and diversity. We found significant differences between the microbiota profiles obtained from each pipeline. At domain level, bacteria were relatively more abundant using QIIME2 than MG-RAST; at phylum level, seven taxa were identified exclusively by QIIME2; at family level, samples processed in QIIME2 showed higher evenness and richness (assessed by Shannon and Simpson indices). The genus-level compositions obtained from each pipeline were used in partial least squares-discriminant analyses (PLS-DA) to discriminate between sample collection sites (caecum, colon and faeces). The results showed that different genera were found to be significant for the models, based on the Variable Importance in Projection, e.g. when using sequencing data processed by MG-RAST, the three most important genera were Acetitomaculum, Ruminococcus and Methanosphaera, whereas when data was processed using QIIME2, these were Candidatus Methanomethylophilus, Sphaerochaeta and Anaerorhabdus. Furthermore, the application of differential filtering procedures before the PLS-DA revealed higher accuracy when using non-restricted datasets obtained from MG-RAST, whereas datasets obtained from QIIME2 resulted in more accurate discrimination of sample collection sites after removing genera with low relative abundances (<1%) from the datasets. Our results highlight the differences in taxonomic compositions of samples obtained from the two separate pipelines, while underlining the impact on downstream analyses, such as biomarkers identification.

Nuclear Magnetic Resonance to Detect Rumen Metabolites Associated with Enteric Methane Emissions from Beef Cattle.

This study presents the application of metabolomics to evaluate changes in the rumen metabolites of beef cattle fed with three different diet types: forage-rich, mixed and concentrate-rich. Rumen fluid samples were analysed by 1H-NMR spectroscopy and the resulting spectra were used to characterise and compare metabolomic profiles between diet types and assess the potential for NMR metabolite signals to be used as proxies of methane emissions (CH4 in g/kg DMI). The dataset available consisted of 128 measurements taken from 4 experiments with CH4 measurements taken in respiration chambers. Predictive modelling of CH4 was conducted by partial least squares (PLS) regression, fitting calibration models either using metabolite signals only as predictors or using metabolite signals as well as other diet and animal covariates (DMI, ME, weight, BW0.75, DMI/BW0.75). Cross-validated R2 were 0.57 and 0.70 for the two models respectively. The cattle offered the concentrate-rich diet showed increases in alanine, valerate, propionate, glucose, tyrosine, proline and isoleucine. Lower methane yield was associated with the concentrate-rich diet (p < 0.001). The results provided new insight into the relationship between rumen metabolites, CH4 production and diets, as well as showing that metabolites alone have an acceptable association with the variation in CH4 production from beef cattle.

A parsimonious software sensor for estimating the individual dynamic pattern of methane emissions from cattle.

Large efforts have been deployed in developing methods to estimate methane emissions from cattle. For large scale applications, accurate and inexpensive methane predictors are required. Within a livestock precision farming context, the objective of this work was to integrate real-time data on animal feeding behaviour with an in silico model for predicting the individual dynamic pattern of methane emission in cattle. The integration of real-time data with a mathematical model to predict variables that are not directly measured constitutes a software sensor. We developed a dynamic parsimonious grey-box model that uses as predictor variables either dry matter intake (DMI) or the intake time (IT). The model is described by ordinary differential equations.Model building was supported by experimental data of methane emissions from respiration chambers. The data set comes from a study with finishing beef steers (cross-bred Charolais and purebred Luing finishing). Dry matter intake and IT were recorded using feed bins. For research purposes, in this work, our software sensor operated off-line. That is, the predictor variables (DMI, IT) were extracted from the recorded data (rather than from an on-line sensor). A total of 37 individual dynamic patterns of methane production were analyzed. Model performance was assessed by concordance analysis between the predicted methane output and the methane measured in respiration chambers. The model predictors DMI and IT performed similarly with a Lin's concordance correlation coefficient (CCC) of 0.78 on average. When predicting the daily methane production, the CCC was 0.99 for both DMI and IT predictors. Consequently, on the basis of concordance analysis, our model performs very well compared with reported literature results for methane proxies and predictive models. As IT measurements are easier to obtain than DMI measurements, this study suggests that a software sensor that integrates our in silico model with a real-time sensor providing accurate IT measurements is a viable solution for predicting methane output in a large scale context.

Nitrogen isotopic fractionation as a biomarker for nitrogen use efficiency in ruminants: a meta-analysis.

Animal proteins are naturally 15N enriched relative to the diet and the extent of this difference (Δ15Nanimal-diet or N isotopic fractionation) has been correlated to N use efficiency (NUE; N gain or milk N yield/N intake) in some recent ruminant studies. The present study used meta-analysis to investigate whether Δ15Nanimal-diet can be used as a predictor of NUE across a range of dietary conditions, particularly at the level of between-animal variation. An additional objective was to identify variables related to N partitioning explaining the link between NUE and Δ15Nanimal-diet. Individual values from eight publications reporting both NUE and Δ15Nanimal-diet for domestic ruminants were used to create a database comprising 11 experimental studies, 41 treatments and individual animal values for NUE (n=226) and Δ15Nanimal-diet (n=291). Data were analyzed by mixed-effect regression analysis taking into account experimental factors as random effects on both the intercept and slope of the model. Diets were characterized according to the INRA feeding system in terms of N utilization at the rumen, digestive and metabolic levels. These variables were used in a partial least squares regression analysis to predict separately NUE and Δ15Nanimal-diet variation, with the objective of identifying common variables linking NUE and Δ15Nanimal-diet. For individuals reared under similar conditions (within-study) and at the same time (within-period), the variance of NUE and Δ15Nanimal-diet not explained by dietary treatments (i.e. between-animal variation plus experimental error) was 35% and 55%, respectively. Mixed-effect regression analysis conducted with treatment means showed that Δ15Nanimal-diet was significantly and negatively correlated to NUE variation across diets (NUE=0.415 -0.055×Δ15Nanimal-diet). When using individual values and taking into account the random effects of study, period and diet, the relationship was also significant (NUE=0.358 -0.035×Δ15Nanimal-diet). However, there may be a biased prediction for animals close to zero, or in negative, N balance. When using a novel statistical approach, attempting to regress between-animal variation in NUE on between-animal variation in Δ15Nanimal-diet (without the influence of experimental factors), the negative relationship was still significant, highlighting the ability of Δ15Nanimal-diet to capture individual variability. Among the studied variables related to N utilization, those concerning N efficiency use at the metabolic level contributed most to predict both Δ15Nanimal-diet and NUE variation, with rumen fermentation and digestion contributing to a lesser extent. This study confirmed that on average Δ15Nanimal-diet can predict NUE variation across diets and across individuals reared under similar conditions.

Apparent recovery of duodenal odd- and branched-chain fatty acids in milk of dairy cows.

This study compared flows of odd- and branched-chain fatty acids (OBCFA) at the duodenum with corresponding yields in milk. Four mid-lactation Holstein-Friesian dairy cows were offered 4 dietary treatments, based on different ratios of ryegrass silage and concentrates (80:20, 65:35, 50:50, and 35:65 on a dry matter basis), in a 4 x 4 Latin square design experiment with 4-wk periods. Samples of milk and duodenal digesta were collected during the final week of each period and analyzed for fatty acids. Biohydrogenation of linoleic and alpha-linolenic acids (C18:2 and C18:3) was extensive for all treatments, with a tendency to be lower for C18:3 with increased concentrate feeding. The proportion of duodenal flows of these fatty acids that appeared in milk declined with increasing concentrate feeding. There was little change in the yield of OBCFA in milk in response to increasing level of concentrate inclusion and no significant relationship with the yield of microbial protein at the duodenum. The efficiency of transfer of iso C15:0 and anteiso C15:0 from the duodenum to milk was similar to that for C18:3, with a reduced proportion transferred into milk at higher flows. Yields of C15:0, C17:0, and iso C17:0 in milk were higher than duodenal flows, confirming synthesis in animal tissues.

Effects of fatty acid oxidation products (green odor) on rumen bacterial populations and lipid metabolism in vitro.

This study investigated the effects of green odor fatty acid oxidation products (FAOP) from cut grass on lipid metabolism and microbial ecology using in vitro incubations of rumen microorganisms. These compounds have antimicrobial roles in plant defense, and we hypothesized that they may influence rumen lipid metabolism. Further, they may partially explain the higher levels of conjugated linoleic acid cis-9, trans-11 in milk from cows grazing pasture. The first of 2 batch culture experiments screened 6 FAOP (1 hydroperoxide, 3 aldehydes, 1 ketone, and 1 alcohol) for effects on lipid profile, and in particular C(18) polyunsaturated fatty acid biohydrogenation. Experiment 2 used the most potent FAOP to determine effects of varying concentrations and identify relationships with effects on microbial ecology. Batch cultures contained anaerobic buffer, rumen liquor, and FAOP to a final concentration of 100 microM for experiment 1. Triplicates for each compound and controls (water addition) were incubated at 39 degrees C for 6 h. The hydroperoxide (1,2-dimethylethyl hydroperoxide, 1,2-DMEH) and the long chain aldehyde (trans-2 decenal) had the largest effects on lipid metabolism with significant increases in C(18:0) and C(18:1) trans and reductions in C(12:0), C(14:0), C(16:0), C(18:1) cis, C(18:2n-6), C(18:3n-3), C(20:0) and total branch and odd chain fatty acids compared with the control. This was associated with significantly higher biohydrogenation of C(18) polyunsaturated fatty acid. In experiment 2, 1,2-DMEH was incubated at 50, 100, and 200 microM for 2, 6, and 24 h. Increasing 1,2-DMEH concentration resulted in a significant linear increase in C(18:1) trans-10, trans-11, conjugated linoleic acid, and C(18:0) and a linear decrease in C(18:2n-6) and C(18:3n-3), although the scale of this response declined with time. Microbial profiling techniques showed that 1,2-DMEH at concentrations of 100 and 200 microM changed the microbial community from as early as 2 h after addition, though microbial biomass remained similar. These preliminary studies have shown that FAOP can alter fatty acid biohydrogenation in the rumen. This change was associated with changes in the microbial population that were detected through DNA and branched- and odd-chain fatty acid profiling approaches.

Effects of dietary protein and starch on intake, milk production, and milk fatty acid profiles of dairy cows fed corn silage-based diets.

Feed intake, milk production, and milk fatty acid profiles of dairy cows fed corn silage-based diets with different protein and starch concentrations were measured in a 3-period experiment in a changeover design using 12 Holstein cows. Each experimental period lasted for 3 wk. The diet fed as a total mixed ration consisted of 45% corn silage, 5% coarsely chopped wheat straw, and 50% concentrate, on a dry matter (DM) basis. The 4 treatments, formulated to be isoenergetic and to differ in concentrations of dietary crude protein (CP) and starch (DM basis), were as follows: low CP and low starch (LPLS; 14% CP and 15% starch), low CP and high starch (LPHS; 14% CP and 25% starch), high CP and low starch (HPLS; 16% CP and 15% starch), and high CP and high starch (HPHS; 16% CP and 25% starch). The LPLS treatment led to lower DM intake, milk yield, milk protein concentration, and milk lactose yield, probably due to a shortage of both rumen-degradable protein supply to rumen microbes and glucogenic nutrients to the animal. There were no differences between protein-rich diets and LPHS, suggesting that this diet satisfied the rumen-degradable protein requirements of rumen microbes and did not limit feed intake, and the increased supply of glucogenic nutrients spared AA so that the nutrient requirements of mid lactation dairy cows were met. Further increases in CP concentration increased plasma urea concentration and resulted in decreased efficiency of conversion of dietary N into milk N. Milk fatty acid profiles were affected by starch and protein supply, with starch having the largest effect. Additionally, increasing dietary starch concentration decreased the apparent transfer of dietary polyunsaturated fatty acids to milk, suggesting an increased channeling of fatty acids to adipose tissue. The results further suggest that C(15:0) and C(17:0) are synthesized de novo in animal tissues.

Current available strategies to mitigate greenhouse gas emissions in livestock systems: an animal welfare perspective.

Livestock production is a major contributor to greenhouse gas (GHG) emissions, so will play a significant role in the mitigation effort. Recent literature highlights different strategies to mitigate GHG emissions in the livestock sector. Animal welfare is a criterion of sustainability and any strategy designed to reduce the carbon footprint of livestock production should consider animal welfare amongst other sustainability metrics. We discuss and tabulate the likely relationships and trade-offs between the GHG mitigation potential of mitigation strategies and their welfare consequences, focusing on ruminant species and on cattle in particular. The major livestock GHG mitigation strategies were classified according to their mitigation approach as reducing total emissions (inhibiting methane production in the rumen), or reducing emissions intensity (Ei; reducing CH4 per output unit without directly targeting methanogenesis). Strategies classified as antimethanogenic included chemical inhibitors, electron acceptors (i.e. nitrates), ionophores (i.e. Monensin) and dietary lipids. Increasing diet digestibility, intensive housing, improving health and welfare, increasing reproductive efficiency and breeding for higher productivity were categorized as strategies that reduce Ei. Strategies that increase productivity are very promising ways to reduce the livestock carbon footprint, though in intensive systems this is likely to be achieved at the cost of welfare. Other strategies can effectively reduce GHG emissions whilst simultaneously improving animal welfare (e.g. feed supplementation or improving health). These win-win strategies should be strongly supported as they address both environmental and ethical sustainability. In order to identify the most cost-effective measures for improving environmental sustainability of livestock production, the consequences of current and future strategies for animal welfare must be scrutinized and contrasted against their effectiveness in mitigating climate change.

Compositional mixed modeling of methane emissions and ruminal volatile fatty acids from individual cattle and multiple experiments.

The aim of the study was to investigate the association of methane (CH) yields (g/kg DMI) with rumen VFA molar proportions and animal and diet-related covariates from individual animals and multiple experiments. The dataset available consisted of 284 measurements of CH yields for beef cattle from 6 experiments measured in indirect respiration chambers. A compositional modeling approach was employed where VFA measurements were considered as a whole, instead of in isolation, emphasizing their multivariate relative scale. The analysis revealed expected close groupings of acetate and butyrate; propionate and valerate; iso-butyrate and iso-valerate. Linear mixed models were then fitted to examine relationships between CH yield and VFA, represented by meaningful log-contrasts of components called compositional balances, while accounting for other animal and diet-related covariates and random variability between experiments. A compositional balance representing (acetate × butyrate)/propionate best explained the contribution of VFA to variation in CH yield. The covariates DMI, forage:concentrate proportion (expressed as a categorical variable diet type: high concentrate, mixed forage:concentrate or high forage), and diet ME were also statistically significant. These results provided new insights into the relative inter-relationships among VFA measurements and also between VFA and CH yield. In conclusion, VFA molar proportions as represented by compositional balances were a significant contributor to explaining variation in CH yields from individual cattle.