Dairy Cows Offered Fresh Chicory Instead of Ensiled Pasture during an Acute Heat Challenge Produced More Milk and Had Lower Body Temperatures.

The frequency, duration, and intensity of heat waves in Australia are increasing. To reduce the impact of heat waves on milk production, novel management strategies are required. Altering the forage type and amount offered affect the heat load on dairy cows and offer potential strategies to ameliorate the effects of hot weather. Thirty-two multiparous, lactating Holstein-Friesian cows were assigned one of four dietary treatments: chicory high amount, chicory low amount, pasture silage high amount, or pasture silage low amount. These cows were exposed to a heat wave in controlled-environment chambers. Cows that were offered fresh chicory had similar feed intake to cows that were offered pasture silage (15.3 kg DM/d). However, cows that were offered chicory produced greater energy-corrected milk (21.9 vs. 17.2 kg/d) and had a lower maximum body temperature (39.4 vs. 39.6 °C) than cows that were offered pasture silage overall. Cows that were offered the high amount of forage had greater feed intake (16.5 vs. 14.1 kg DM/d) and energy corrected milk yield (20.0 vs. 17.9 kg/d) than cows that were offered the low amount, as intended, but with no difference in maximum body temperature (39.5 °C). We conclude that feeding chicory instead of pasture silage to dairy cows shows promise as a dietary strategy to ameliorate the effect of heat exposure, and there was no advantage in restricting feed amount.

Effects of Heat Stress in Dairy Cows Offered Diets Containing Either Wheat or Corn Grain during Late Lactation.

Cereal grains that differ in the rate and extent of ruminal fermentation differ in heat increment and may be used to improve thermoregulation during heat stress. This experiment investigated the responses of dairy cows in late lactation to a heat challenge when offered wheat-grain or corn-grain. Eighteen lactating cows, 220 ± 94 (mean ± standard deviation) days in milk, 3.7 ± 0.17 years of age and 558 ± 37 kg bodyweight, were allocated treatments containing 6 kg dry matter (DM)/day of wheat grain or 6 kg DM/day corn grain (9 per treatment) plus 14 kg DM/day of alfalfa hay. Measurements were made during a 7-day pre-challenge period at ambient conditions in individual stalls, during a 4-day heat challenge (temperature humidity index of 74 to 84) in individual controlled-climate chambers, then during a 7-day recovery period at ambient conditions in individual stalls. During the heat challenge, cows offered corn had lower respiration rates (p = 0.017) and greater feed intake (p = 0.021) but energy-corrected milk (p = 0.097) was not different to that of cows offered wheat. Feeding corn grain to dairy cows during a heat challenge reduced some of the negative impacts of heat stress, enabling the cows to consume more forage compared with supplementing with wheat grain.

Dietary wheat and reduced methane yield are linked to rumen microbiome changes in dairy cows.

Fermentation of pasture grasses and grains in the rumen of dairy cows and other ruminants produces methane as a by-product, wasting energy and contributing to the atmospheric load of greenhouse gasses. Many feeding trials in farmed ruminants have tested the impact of dietary components on feed efficiency, productivity and methane yield (MeY). Such diets remodel the rumen microbiome, altering bacterial, archaeal, fungal and protozoan populations, with an altered fermentation outcome. In dairy cows, some dietary grains can reduce enteric methane production. This is especially true of wheat, in comparison to corn or barley. Using a feeding trial of cows fed rolled wheat, corn or barley grain, in combination with hay and canola, we identified wheat-associated changes in the ruminal microbiome. Ruminal methane production, pH and VFA concentration data together with 16S rRNA gene amplicon sequences were used to compare ruminal bacterial and archaeal populations across diets. Differential abundance analysis of clustered sequences (OTU) identified members of the bacterial families Lachnospiraceae, Acidaminococcaceae, Eubacteriaceae, Prevotellaceae, Selenomonadaceae, Anaerovoracaceae and Fibrobacteraceae having a strong preference for growth in wheat-fed cows. Within the methanogenic archaea, (at >99% 16S rRNA sequence identity) the growth of Methanobrevibacter millerae was favoured by the non-wheat diets, while Methanobrevibacter olleyae was unaffected. From the wheat-preferring bacteria, correlation analysis found OTU strongly linked to reduced MeY, reduced pH and raised propionic acid levels. OTU from the genera Shuttleworthia and Prevotella_7 and especially Selenomonadaceae had high anti-methane correlations. An OTU likely representing (100% sequence identity) the fumarate-reducing, hydrogen-utilising, rumen bacterium Mitsuokella jalaludinii, had an especially high negative correlation coefficient (-0.83) versus MeY and moderate correlation (-0.6) with rumen pH, strongly suggesting much of the MeY suppression is due to reduced hydrogen availablity. Other OTU, representing as yet unknown species from the Selenomonadaceae family and the genera Prevotella_7, Fibrobacter and Syntrophococcus also had high to moderate negative MeY correlations, but low correlation with pH. These latter likely represent bacterial species able to reduce MeY without causing greater ruminal acidity, making them excellent candidates, provided they can be isolated, for development as anti-methane probiotics.

Dietary Fat and Betaine Supplements Offered to Lactating Cows Affect Dry Matter Intake, Milk Production and Body Temperature Responses to an Acute Heat Challenge.

Supplementing the diet of lactating cows with ingredients that increase energy density, or reduce internal heat production, may reduce some of the negative impacts of hot weather on milk yield. Thirty-two dairy cows were assigned either: (1) basal diet only, (2) basal diet plus canola oil, (3) basal diet plus betaine, or (4) basal diet plus canola oil and betaine. The basal diet was lucerne hay, pasture silage, and grain. Cows were exposed to a four-day heat challenge (temperature-humidity index 74 to 84) in controlled-environment chambers. Canola oil supplementation increased milk production (22.0 vs. 18.7 kg/d) across all periods of our experiment and increased body temperature (39.6 vs. 39.0 °C) during the heat challenge. Betaine supplementation reduced maximum body temperature during the pre-challenge period (39.2 vs. 39.6 °C) but not during the heat challenge (40.3 °C). Cows fed canola oil had greater declines in dry matter intake (5.4 vs 2.7 kg DM) and energy corrected milk (1.3 vs. 1.0 kg) from the pre-challenge to the heat challenge than other cows. Contrary to our expectations, the combination of fat and betaine supplements did not result in a clear benefit in terms of milk production or body temperature. Further work is warranted to understand the interactions between diet and hot weather.

A meta-analysis of effects of dietary seaweed on beef and dairy cattle performance and methane yield.

There has been considerable interest in the use of red seaweed, and in particular Asparagopsis taxiformis, to increase production of cattle and to reduce greenhouse gas emissions. We hypothesized that feeding seaweed or seaweed derived products would increase beef or dairy cattle performance as indicated by average daily gain (ADG), feed efficiency measures, milk production, and milk constituents, and reduce methane emissions. We used meta-analytical methods to evaluate these hypotheses. A comprehensive search of Google Scholar, Pubmed and ISI Web of Science produced 14 experiments from which 23 comparisons of treatment effects could be evaluated. Red seaweed (Asparagopsis taxiformis) and brown seaweed (Ascophyllum nodosum) were the dominant seaweeds used. There were no effects of treatment on ADG or dry matter intake (DMI). While there was an increase in efficiency for feed to gain by 0.38 kg per kg [standardized mean difference (SMD) = 0.56; P = 0.001] on DerSimonian and Laird (D&L) evaluation, neither outcome was significant using the more rigorous robust regression analysis (P >0.06). The type of seaweed used was not a significant covariable for ADG and DMI, but A. nodosum fed cattle had lesser feed to gains efficiency compared to those fed A. taxiformis. Milk production was increased with treatment on weighted mean difference (WMD; 1.35 ± 0.44 kg/d; P <0.001); however, the SMD of 0.45 was not significant (P = 0.111). Extremely limited data suggest the possibility of increased percentages of milk fat (P = 0.040) and milk protein (P = 0.001) on (D&L) WMD evaluation. The limited data available indicate dietary supplementation with seaweed produced a significant and substantial reduction in methane yield by 5.28 ± 3.5 g/kg DMI (P = 0.003) on D&L WMD evaluation and a D&L SMD of -1.70 (P = 0.001); however, there was marked heterogeneity in the results (I2 > 80%). In one comparison, methane yield was reduced by 97%. We conclude that while there was evidence of potential for benefit from seaweed use to improve production and reduce methane yield more in vivo experiments are required to strengthen the evidence of effect and identify sources of heterogeneity in methane response, while practical applications and potential risks are evaluated for seaweed use.

Measurement of Enteric Methane Emissions by the SF6 Technique Is Not Affected by Ambient Weather Conditions.

Despite the fact that the sulphur hexafluoride (SF6) tracer technique was developed over 25 years ago to measure methane production from grazing and non-housed animals, no studies have specifically investigated whether ambient wind speed, temperature, relative humidity and rainfall influence the accuracy of the method. The aim of this research was to investigate how these weather factors influence the measurement of enteric methane production by the SF6 technique. Six different cohorts of dairy cows (40 per cohort) were kept outdoors and fed a common diet during spring in 3 consecutive years. Methane production from individual cows was measured daily over the last 5 days of each 32-day period. An automated weather station measured air temperature, wind speed, relative humidity and rainfall every 10 min. Regression analyses were used to relate the average daily wind speed, average daily temperature, average daily relative humidity and total daily rainfall measurements to dry matter intake, average daily methane production and methane yield of each cohort of cows. It was concluded that the modified SF6 technique can be used outdoors during a range of wind speeds, ambient temperatures, relative humidities and rainfall conditions without causing a significant effect on the measurement of methane production or methane yield of dairy cows.

Economic Threshold Analysis of Supplementing Dairy Cow Diets with Betaine and Fat during a Heat Challenge: A Pre- and Post-Experimental Comparison.

Ex ante economic analysis can be used to establish the production threshold for a proposed experimental diet to be as profitable as the control treatment. This study reports (1) a pre-experimental economic analysis to estimate the milk production thresholds for an experiment where dietary supplements were fed to dairy cows experiencing a heat challenge, and (2) comparison of these thresholds to the milk production results of the subsequent animal experiment. The pre-experimental thresholds equated to a 1% increase in milk production for the betaine supplement, 9% increase for the fat supplement, and 11% increase for fat and betaine in combination, to achieve the same contribution to farm profit as the control diet. For the post-experimental comparison, previously modelled climate predictions were used to extrapolate the milk production results from the animal experiment over the annual hot-weather period for the dairying region in northern Victoria, Australia. Supplementing diets with fat or betaine had the potential to produce enough extra milk to exceed the production thresholds, making either supplement a profitable alternative to feeding the control diet during the hot-weather period. Feeding fat and betaine in combination failed to result in the extra milk required to justify the additional cost when compared to the control diet.

Ozone Decreased Enteric Methane Production by 20% in an in vitro Rumen Fermentation System.

Ozone (O3) is volatile, highly oxidative, and has theoretical potential to reduce ruminant enteric methanogenesis by interactions between archaea and bacteria, and substrate and oxygen. The effects of O3 on the rumen microbiota, fermentation parameters, and CH4 emissions were studied through in vitro fermentation using a RUSITEC apparatus with O3 dissolved in the salivary buffer. The substrate consisted of maize silage or grain concentrates, and the treatments were (1) control (no O3) and (2) O3 at 0.07 ± 0.022 mg/L in the buffer. A 4-day adaptation period followed by a 6-day experimental period was used for measuring gas production and composition, as well as fermentation characteristics, which included ruminal volatile fatty acids (VFA) and liquid- and solid-associated microbial communities. Ozone treatment decreased total gas production by 15.4%, most notably CH4 production by 20.4%, and CH4 gas concentration by 5.8%, without compromising dry matter digestibility (DMD) of either maize silage or grain concentrates. There were no significant effects of O3 treatment on VFA production or pH. Ozone treatment reduced the relative abundance of methanogens, particularly Methanomicrobium. This study demonstrates the potential use of O3 as a method to reduce ruminant enteric methanogenesis.

An alternative approach for sustainable sheep meat production: implications for food security.

BACKGROUND: A pelleted diet containing camelina hay (CAMH) or camelina meal (CAMM) as a supplement along with a control pellet (CONT) diet formulated with commonly available feeds during summer was used to investigate an alternative pathway for sustainable meat production. Sustainable meat production was based on a simple estimation of income from meat produced versus feed costs if animals were fed for an extended period over summer compared to early slaughter at the beginning of summer. Eighty maternal composite wether lambs (Composite) based on Coopworth genetics and 80 pure Merino wether yearlings were divided into 10 groups within breed (n = 8) using stratified randomisation based on liveweights. Following 1 week of adaptation to experimental diets, animals were fed experimental diets for up to10 weeks. RESULTS: Animals were slaughtered after either 8, 9 or 10 weeks of full feeding when the average liveweight of diet/genetic combination reached a weight appropriate for either 'heavy lamb' or 'heavy hogget' production, which occurred between 8 and 10 weeks of full feeding. There was no diet × breed interactions except for dressing percentage (DP), where Composite lambs fed the CAMH diet had the greatest DP (48.1 ± 0.35) and the Merino yearlings fed the CAMM diet the lowest DP (45.8 ± 0.33). Composite lambs gained 17.6-20.3 kg and Merino yearlings gained 10.7-12.9 kg liveweight. Based on their DP, this resulted in the production of approximately 8.3-9.5 kg additional carcass weight in Composites and 4.9-5.7 kg in Merinos, which in turn produced greater profit per Composite lamb and a small profit per Merino yearling. CONCLUSIONS: Composite lambs fed CAMM and CAMH had 5% greater carcass weights at slaughter compared to the CONT group, but dietary treatments did not change carcass weight of Merino yearlings at slaughter. The extended feeding approach offered the producer an estimated economic gain of AUD $20.00 to $25.00 when yearly average prices were used (Method 1) and AUD $40.00 to $47.70 when pre- and post-summer average prices were used (Method 2) per Composite lambs, but extra carcass gain did not result in the same profit per Merino yearling. Among the Composites, the profit for animals fed the CAMH and CAMM were AUD $2.75 to $4.50 greater than CONT group when full year average prices were applied while AUD $3.50 to $5.50 greater than CONT group when pre- and post-summer average prices were applied. However, we acknowledge a combination approach of extended feeding for a portion of animals already on ground and selling the remaining animals pre-summer with joining of additional ewes is the most likely strategy. Considering the scenario of extended feeding for 3 weeks, based on the growth rates observed for Composite lambs, if an additional 2 kg carcass weight per animal can be gained for 50% of the 22 million lambs slaughtered in Australia (= 11 million animals), it would potentially supply an additional 22 million kg of lamb carcasses produced per annum. This is equivalent to producing an extra 1 million lamb carcasses per annum weighing 22 kg each. Feeding Composite lambs for an extended period and selling Merino yearlings pre-summer may be a good option due to faster growth rate of Composites that may help quick turn-over to market.

Effects of Feeding either Red or White Grape Marc on Milk Production and Methane Emissions from Early-Lactation Dairy Cows.

Globally, annual production of grape marc (GM), the residue of skins, seeds and stems remaining after making wine, has been estimated to be approximately nine million tonnes. No previous studies have compared effects on milk production and methane emissions when GM from either red or white grapes was fed to dairy cows. This experiment examines the effects of partial replacement of a perennial ryegrass (Lolium perenne L.) based diet with GM from either red or white grapes on yield and composition of milk and methane emissions. Thirty-two Holstein dairy cows in early lactation were offered either a control diet containing 15.0 kg dry matter (DM) of freshly harvested perennial ryegrass and 5.2 kg of a concentrate mix, or a diet similar to the control diet but with 5 kg DM of ryegrass replaced with 5 kg DM of GM from red grapes (RGM), or a diet similar to the RGM diet except the GM was from white grapes (WGM). Individual cow feed intakes, milk yields, and methane emissions were measured. Both diets containing GM decreased milk yields by approximately 10% and methane emissions by 15%. When fed to dairy cows, GM reduces methane emissions but at the cost of decreased milk production.

Volatile Fatty Acids in Ruminal Fluid Can Be Used to Predict Methane Yield of Dairy Cows.

The dry matter intake (DMI) of forage-fed cattle can be used to predict their methane emissions. However, many cattle are fed concentrate-rich diets that decrease their methane yield. A range of equations predicting methane yield exist, but most use information that is generally unavailable when animals are fed in groups or grazing. The aim of this research was to develop equations based on proportions of ruminal volatile-fatty-acids to predict methane yield of dairy cows fed forage-dominant as well as concentrate-rich diets. Data were collated from seven experiments with a total of 24 treatments, from 215 cows. Forage in the diets ranged from 440 to 1000 g/kg. Methane was measured either by open-circuit respiration chambers or a sulfur hexafluoride (SF6) technique. In all experiments, ruminal fluid was collected via the mouth approximately four hours after the start of feeding. Seven prediction equations were tested. Methane yield (MY) was equally best predicted by the following equations: MY = 4.08 × (acetate/propionate) + 7.05; MY = 3.28 × (acetate + butyrate)/propionate + 7.6; MY = 316/propionate + 4.4. These equations were validated against independent published data from both dairy and beef cattle consuming a wide range of diets. A concordance of 0.62 suggests these equations may be applicable for predicting methane yield from all cattle and not just dairy cows, with root mean-square error of prediction of 3.0 g CH4/kg dry matter intake.

The effect of diet of the donor cows on in vitro measurements of methane production from wheat and corn incubated in various forage-to-grain ratios.

BACKGROUND: Supplementation of ruminant diets with wheat and corn grains influences ruminal fermentation. In vitro fermentation is a methodology that can be used to screen feeds for their potential to produce enteric methane. However, there is evidence that the diet of the donor cows could impact the results of in vitro analysis. This research investigated the in vitro fermentation of wheat and corn grain when incubated in ruminal fluid from cows fed different grain types and different forage-to-grain ratios. RESULTS: The type of grain fed to the donor cows, as well as forage-to-grain ratio, affected the outcome of fermentation of wheat and corn grain. Differences in methane production (MP) between grains were only observed when incubated with ruminal fluid adapted to each specific grain type. Increasing proportions of wheat but not of corn decreased in vitro MP in a linear manner compared with MP produced from forage only. CONCLUSIONS: Wheat grain has a greater in vitro antimethanogenic effect than corn. However, to detect the different fermentations between wheat and corn, grains should be incubated in ruminal fluid from cows adapted to that specific grain type. © 2019 Society of Chemical Industry.

Comparison of five methods for the estimation of methane production from vented in vitro systems.

BACKGROUND: There are several methods for estimating methane production (MP) from feedstuffs in vented in vitro systems. One method (A; 'gold standard') measures methane proportions in the incubation bottle's headspace (HS) and in the vented gas collected in gas bags. Four other methods (B, C, D and E) measure methane proportion in a single gas sample from the HS. Method B assumes the same methane proportion in the vented gas as in the HS, method C assumes constant methane to carbon dioxide ratio, method D has been developed based on empirical data, and method E assumes constant individual venting volumes. This study aimed to compare the MP predictions from these methods to that of the gold standard method under different incubation scenarios, to validate these methods based on their concordance with a gold-standard method. RESULTS: Methods C, D and E had greater concordance (0.85, 0.88 and 0.81), lower root-mean-square error (RMSE; 0.80, 0.72 and 0.85) and lower mean bias (0.20, 0.35, -0.35) with the gold standard than did method B (concordance 0.67, RMSE 1.49 and mean bias 1.26). Methods D and E were simpler to perform than method C, and method D was slightly more accurate than method E. CONCLUSION: Based on precision, accuracy and simplicity of implementation, it is recommended that, when method A cannot be used, methods D and E are preferred to estimate MP from vented in vitro systems. © 2018 Society of Chemical Industry.

Influence of acid hydrolysis, saponification and sample clean-up on the measurement of phytosterols in dairy cattle feed using GC-MS and GC with flame ionization detection.

The fortification of processed foods including dairy products is increasingly commonplace with phytosterols among many compounds used to improve the nutritional value of food products. It is also increasingly common practice for some dairy cattle feeds to be fortified for their potential to increase phytosterol levels in milk. In this study, a combined, streamlined protocol using acid hydrolysis, saponification and sample clean-up was developed to enable the rapid and reliable measurement of phytosterols. The method was developed with focus on streamlining the overall technique to make it suitable for commercial laboratories, to reduce labor and consumable costs, while maintaining accuracy. A total of 12 different feed types commonly used in the dairy industry were analyzed with the highest and lowest sterol contents found in cotton seed oil and tannin with average phytosterol contents of 256 and <30 mg per 100 g, respectively. With a limit of reporting of 30 mg/kg for individual sterols and a correlation coefficient > 0.99, the method was validated for milk to enable feed comparison studies with respect to the total phytosterol content in raw milk.

Prediction of enteric methane production, yield, and intensity in dairy cattle using an intercontinental database.

Enteric methane (CH4 ) production from cattle contributes to global greenhouse gas emissions. Measurement of enteric CH4 is complex, expensive, and impractical at large scales; therefore, models are commonly used to predict CH4 production. However, building robust prediction models requires extensive data from animals under different management systems worldwide. The objectives of this study were to (1) collate a global database of enteric CH4 production from individual lactating dairy cattle; (2) determine the availability of key variables for predicting enteric CH4 production (g/day per cow), yield [g/kg dry matter intake (DMI)], and intensity (g/kg energy corrected milk) and their respective relationships; (3) develop intercontinental and regional models and cross-validate their performance; and (4) assess the trade-off between availability of on-farm inputs and CH4 prediction accuracy. The intercontinental database covered Europe (EU), the United States (US), and Australia (AU). A sequential approach was taken by incrementally adding key variables to develop models with increasing complexity. Methane emissions were predicted by fitting linear mixed models. Within model categories, an intercontinental model with the most available independent variables performed best with root mean square prediction error (RMSPE) as a percentage of mean observed value of 16.6%, 14.7%, and 19.8% for intercontinental, EU, and United States regions, respectively. Less complex models requiring only DMI had predictive ability comparable to complex models. Enteric CH4 production, yield, and intensity prediction models developed on an intercontinental basis had similar performance across regions, however, intercepts and slopes were different with implications for prediction. Revised CH4 emission conversion factors for specific regions are required to improve CH4 production estimates in national inventories. In conclusion, information on DMI is required for good prediction, and other factors such as dietary neutral detergent fiber (NDF) concentration, improve the prediction. For enteric CH4 yield and intensity prediction, information on milk yield and composition is required for better estimation.

Modelling the Effect of Diet Composition on Enteric Methane Emissions across Sheep, Beef Cattle and Dairy Cows.

Enteric methane (CH 4 ) is a by-product from fermentation of feed consumed by ruminants, which represents a nutritional loss and is also considered a contributor to climate change. The aim of this research was to use individual animal data from 17 published experiments that included sheep ( n = 288), beef cattle ( n = 71) and dairy cows ( n = 284) to develop an empirical model to describe enteric CH 4 emissions from both cattle and sheep, and then evaluate the model alongside equations from the literature. Data were obtained from studies in the United Kingdom (UK) and Australia, which measured enteric CH 4 emissions from individual animals in calorimeters. Animals were either fed solely forage or a mixed ration of forage with a compound feed. The feed intake of sheep was restricted to a maintenance amount of 875 g of DM per day (maintenance level), whereas beef cattle and dairy cows were fed to meet their metabolizable energy (ME) requirement (i.e., production level). A linear mixed model approach was used to develop a multiple linear regression model to predict an individual animal's CH 4 yield (g CH 4 /kg dry matter intake) from the composition of its diet. The diet components that had significant effects on CH 4 yield were digestible organic matter (DOMD), ether extract (EE) (both g/kg DM) and feeding level above maintenance intake: CH 4 (g/kg DM intake) = 0.046 (±0.001) × DOMD - 0.113 (±0.023) × EE - 2.47 (±0.29) × (feeding level - 1), with concordance correlation coefficient ( CCC ) = 0.655 and RMSPE = 14.0%. The predictive ability of the model developed was as reliable as other models assessed from the literature. These components can be used to predict effects of diet composition on enteric CH 4 yield from sheep, beef and dairy cattle from feed analysis information.

Identification and quantification of triacylglycerols containing n-3 long-chain polyunsaturated fatty acids in bovine milk.

The n-3 long-chain polyunsaturated fatty acids (LC-PUFA) are low-abundance components in milk fat, but have great potential in promoting human health. A comprehensive survey on triacylglycerol (TAG) molecular species in milk that contain at least one type of n-3 LC-PUFA, namely eicosapentaenoic acid, docosahexaenoic acid, and docosapentaenoic acid, was conducted in this work using HPLC-linear trap quadrupole-Orbitrap and HPLC-triple quadrupole mass spectrometry techniques. A total of 51 TAG species that contain n-3 LC-PUFA have been identified in bovine milk and their structures assigned. The TAG species containing docosahexaenoic acid were found in much smaller number and at much lower abundance compared with the other 2 types of TAG. An HPLC-triple quadrupole mass spectrometry-based method was developed, which provides relative quantification of all these TAG species in a run of 36 min. Application of this method to the quantification of n-3 LC-PUFA-incorporated TAG in 32 individual animal milk samples allowed us to determine variation between animals, identify strong metabolic relationships between TAG species, and reveal negative effect of a grape marc supplement on the accumulation of eicosapentaenoic acid in milk.

An inhibitor persistently decreased enteric methane emission from dairy cows with no negative effect on milk production.

A quarter of all anthropogenic methane emissions in the United States are from enteric fermentation, primarily from ruminant livestock. This study was undertaken to test the effect of a methane inhibitor, 3-nitrooxypropanol (3NOP), on enteric methane emission in lactating Holstein cows. An experiment was conducted using 48 cows in a randomized block design with a 2-wk covariate period and a 12-wk data collection period. Feed intake, milk production, and fiber digestibility were not affected by the inhibitor. Milk protein and lactose yields were increased by 3NOP. Rumen methane emission was linearly decreased by 3NOP, averaging about 30% lower than the control. Methane emission per unit of feed dry matter intake or per unit of energy-corrected milk were also about 30% less for the 3NOP-treated cows. On average, the body weight gain of 3NOP-treated cows was 80% greater than control cows during the 12-wk experiment. The experiment demonstrated that the methane inhibitor 3NOP, applied at 40 to 80 mg/kg feed dry matter, decreased methane emissions from high-producing dairy cows by 30% and increased body weight gain without negatively affecting feed intake or milk production and composition. The inhibitory effect persisted over 12 wk of treatment, thus offering an effective methane mitigation practice for the livestock industries.

Milk Bottom-Up Proteomics: Method Optimization.

Milk is a complex fluid whose proteome displays a diverse set of proteins of high abundance such as caseins and medium to low abundance whey proteins such as ß-lactoglobulin, lactoferrin, immunoglobulins, glycoproteins, peptide hormones, and enzymes. A sample preparation method that enables high reproducibility and throughput is key in reliably identifying proteins present or proteins responding to conditions such as a diet, health or genetics. Using skim milk samples from Jersey and Holstein-Friesian cows, we compared three extraction procedures which have not previously been applied to samples of cows' milk. Method A (urea) involved a simple dilution of the milk in a urea-based buffer, method B (TCA/acetone) involved a trichloroacetic acid (TCA)/acetone precipitation, and method C (methanol/chloroform) involved a tri-phasic partition method in chloroform/methanol solution. Protein assays, SDS-PAGE profiling, and trypsin digestion followed by nanoHPLC-electrospray ionization-tandem mass spectrometry (nLC-ESI-MS/MS) analyses were performed to assess their efficiency. Replicates were used at each analytical step (extraction, digestion, injection) to assess reproducibility. Mass spectrometry (MS) data are available via ProteomeXchange with identifier PXD002529. Overall 186 unique accessions, major and minor proteins, were identified with a combination of methods. Method C (methanol/chloroform) yielded the best resolved SDS-patterns and highest protein recovery rates, method A (urea) yielded the greatest number of accessions, and, of the three procedures, method B (TCA/acetone) was the least compatible of all with a wide range of downstream analytical procedures. Our results also highlighted breed differences between the proteins in milk of Jersey and Holstein-Friesian cows.

Comprehensive polar lipid identification and quantification in milk by liquid chromatography-mass spectrometry.

Polar lipids (PLs) are a significant functional component of milk that are difficult to quantitate. A simple method for comprehensive identification and quantitative analysis of all essential PL species using bovine milk is described. The lipid fraction was extracted by a mix of chloroform and methanol and the extract was directly used for PL identification and quantification. PLs were separated by hydrophilic interaction liquid chromatography (HILIC) and detected by an Orbitrap mass analyser in positive mode. The structure of PLs was established or confirmed by tandem MS in both positive and negative modes. The method is sensitive (with a LOD for all PL classes ≤0.1 ng) and reproducible, enabling simultaneous quantification of 70 PL species within a run of 45 min. Application of this method to the quantification of PLs in 32 bovine milk samples revealed the relative abundance of different PL classes, significant variation of PL content between individual samples and the correlation between the major PL classes. The method provides a tool for investigating the variation and metabolism of important PL components in bovine and human milk and in diverse mammalian species.