Diet-induced milk fat depression (MFD) is a multifactorial condition resulting from the interaction of numerous risk factors, including diet fermentability and unsaturated fatty acids concentration, feed additives, and individual cow effects. 2-Hydroxy-4-(methylthio)butanoate (HMTBa) is a methionine analog that has been observed to increase milk fat in some cases, and interactions with MFD risk factors may exist. The objective was to evaluate the effect of HMTBa supplementation on milk fat synthesis in cows with different levels of milk production and fed diets with increasing risk of biohydrogenation-induced MFD. Sixteen high-producing cows (44.1 ± 4.5 kg of milk/d; mean ± SD) and 14 low-producing (31.4 ± 4.3 kg of milk/d) were used in a randomized block design. Treatments were unsupplemented control and HMTBa fed at 0.1% of diet dry matter (25 g/d at 25 kg of dry matter intake). The experiment was 70 d and included a 14-d covariate period followed by 3 phases whereby diets were fed with increasing risk of MFD to determine the interaction of treatment and diet-induced MFD. During the low-risk phase, the base diet was balanced to 33.5% neutral detergent fiber (NDF) and had no exogenous oil (28 d); during the moderate-risk phase, the diet was balanced to 31% NDF and contained 0.75% soybean oil (14 d); and, during the high-risk phase, the diet was balanced to 28.5% NDF and contained 1.5% soybean oil (14 d). An interaction of treatment, production-level, and dietary phase was observed. Low producing cows neither experienced substantial biohydrogenation-induced MFD nor a response in milk fat to HMTBa supplementation. In high-producing cows, HMTBa maintained higher milk fat concentration during the moderate- (2.94 vs. 3.49%) and high-risk (2.38 vs. 3.11%) phases. High-producing cows receiving HMTBa also had greater milk fat yield (0.94 vs. 1.16 kg/d) and lower trans-10 C18:1 (6.11 vs. 1.50) during the high-risk phase. In conclusion, HMTBa increased milk fat in situations with a high risk of biohydrogenation-induced MFD by decreasing absorption of alternate biohydrogenation intermediates.

Rumen microbial biohydrogenation (BH) of unsaturated fatty acids (UFA) has been extensively studied in vitro; however, in vitro BH pathways, rates, and extents may not parallel those in vivo. The objective was to develop an assay to assess in vivo rates, pathways, and extent of BH of oleic (OA), linoleic (LA), and α-linolenic (ALA) acids. Each UFA was characterized in a separate experiment, each using 4 ruminally cannulated lactating Holstein cows. A single bolus consisting of 200 g of a UFA-oil [experiment 1 (EXP1): 87% OA sunflower, experiment 2 (EXP2): 70% LA safflower, and experiment 3 (EXP3): 54% ALA flaxseed] and 12 g of heptadecanoic acid (C17:0) was mixed into the rumen through the fistula. Rumen digesta was collected at -1, -0.25, 0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, and 6 h relative to the bolus. Overall, the triglyceride boluses increased total fatty acids (FA) in the rumen from 3.9 (standard deviation = ±1.4) to 7.3% (±1.4) of rumen dry matter and enriched C17:0 from 0.4 (±0.1) to 2.5% (±0.5) of FA. The bolus enriched OA from 8.9 (±1.0) to 30.1% (±4.6) of FA in EXP1, LA from 11.1 (±1.8) to 35.9% (±5.0) of FA in EXP2, and ALA from 2.1 (±0.1) to 19.8% (±4.3) of FA in EXP3. The disappearances of C17:0, OA, LA, and ALA were fit to a single exponential decay model. The first-order rate of C17:0 rumen disappearance (turnover) was 9.1, 6.9, and 5.2%/h in EXP1, EXP2, and EXP3, respectively, and was used as a marker of FA passage. The rate of total rumen turnover of OA was 54.1%/h, LA was 60.5%/h, and ALA was 93.0%/h in EXP1, EXP2, and EXP3, respectively. Rumen concentration of all 3 UFA reached prebolus concentrations within 4 h. The calculated extent of lipolysis and initial isomerization was 85.6% for OA, 89.8% for LA, and 94.7% for ALA in EXP1, EXP2, and EXP3, respectively. Assuming that BH equals total disappearance minus passage, the rates of lipolysis and initial isomerization were 45.0, 53.6, and 87.8%/h for OA, LA, and ALA in EXP1, EXP2, and EXP3, respectively. Analysis of the data using compartmental modeling showed that the normal BH pathways proposed in the literature explained 46.0, 37.3, and 49.8% of the BH of OA, LA, and ALA in EXP1, EXP2, and EXP3, respectively. Based on the model, BH of trans C18:1 FA was the rate-limiting step to complete BH. Importantly, oils were provided as triglycerides and the reported rates represent the rate of lipolysis and BH. In conclusion, the rate of ruminal BH of OA, LA, and ALA was higher than that commonly observed in vitro, but the extent of BH was near expected values. The method developed provides a potential in vivo assay of ruminal BH for use in future experiments and modeling efforts.

Diet-induced milk fat depression (MFD) is a condition marked by a reduction in milk fat yield experimentally achieved by increasing dietary unsaturated fatty acids and fermentable carbohydrates. 2-Hydroxy-4-(methylthio) butanoate (HMTBa) is a methionine analog observed to reduce diet-induced MFD in dairy cows. We hypothesize that the reduction in diet-induced MFD by HMTBa is due to changes in the rumen microbiota. To test this, 22 high-producing cannulated Holstein dairy cows were placed into 2 groups using a randomized block design and assigned to either control or HMTBa supplementation (0.1% of diet dry matter). All cows were then exposed to 3 different diets with a low risk (32% neutral detergent fiber, no added oil; fed d 1 to 7), a moderate risk (29% neutral detergent fiber and 0.75% soybean oil; fed d 8 to 24), or a high risk (29% neutral detergent fiber and 1.5% soybean oil; fed d 25 to 28) for diet-induced MFD. Rumen samples were collected on d 0, 14, 24, and 28, extracted for DNA, PCR-amplified for the V1-V2 region of the 16S rRNA gene, sequenced on an Illumina MiSeq (Illumina, San Diego, CA), and subjected to bacterial diversity analysis using the QIIME pipeline. The α diversity estimates (species richness and Shannon diversity) were decreased in the control group compared with the HMTBa group. Bacterial community composition also differed between control and HMTBa groups based on both weighted UniFrac (relative abundance of commonly detected bacteria) and unweighted UniFrac (presence/absence) distances. Within the HMTBa group, no differences were observed in bacterial community composition between d 0 and d 14, 24, and 28; however, in the control group, d 0 samples were different from d 14, 24, and 28. Certain bacterial genera including Dialister, Megasphaera, Lachnospira, and Sharpea were increased in the control group compared with the HMTBa group. Interestingly, these genera were positively correlated with milk fat trans-10,cis-12 conjugated linoleic acid and trans-10 C18:1, fatty acid isomers associated with biohydrogenation-induced MFD. It can be concluded that diet-induced MFD is accompanied by significant alterations in the rumen bacterial community and that HMTBa supplementation reduces these microbial perturbations.

The extent to which CLA supplements inhibit milk fat synthesis is highly dependent on the amount of trans-10, cis-12 CLA reaching the mammary gland. Secondary events such as changes in milk yield, dry matter intake (DMI), body weight (BW) and energy balance (EB) may also occur depending on the magnitude of milk fat depression. This study investigated the effects of feeding a rumen unprotected CLA methyl ester supplement on milk fat yield and fatty acid (FA) profile, yield of milk and other milk components, DMI and metabolic variables in dairy goats. Twenty primiparous, non-pregnant Toggenburg goats, paired by BW, days in milk (DIM) and milk yield (MY) [2.8 ± 0.4 (mean ± SEM) kg milk/day, BW of 40 ± 3.7 kg and 75 ± 3 days in milk] were used in a crossover design with 14 days treatment periods and a 6 days washout. Goats were randomly assigned to the following treatments: Control (30 g/day of calcium salts of FA) or CLA [30 g/day of a rumen unprotected CLA methyl ester supplement (29.9% trans-10, cis-12 CLA)]. There was no treatment effect on BW, milk yield, or milk protein and lactose content and yield. However, compared to Control, CLA decreased milk fat content and yield by 13.9 and 13.3%, respectively, and CLA treated goats consumed 6.5% less corn silage. The CLA treatment increased the concentration of both CLA isomers (cis-9, trans-11 and trans-10, cis-12) in milk fat. The concentration of short and medium chain milk FA (C16) was decreased by CLA, while long chain FA were increased. However, the daily secretion was lower for FA of all chain lengths in CLA treated goats. Glucose and insulin were not affected by treatment, while CLA treatment decreased non esterified FA by 22.2%. The CLA treatment improved calculated EB by 0.3 Mcal/day. Overall, CLA methyl ester supplements are effective in causing milk fat depression in lactating dairy goats and improving EB, which may be used as a nutritional tool in some circumstances.

Feeding trans-10, cis-12 CLA supplements in a rumen-protected form has been shown to cause milk fat depression (MFD) in cows, ewes, and goats. Methyl esters of CLA were shown to be as effective as FFA in inducing MFD when infused postruminally, but their efficacy as a feed supplement has not been addressed in studies with lactating ruminants. In the present study, we investigated the effects of an unprotected trans-10, cis-12 CLA supplement as methyl esters on performance, milk composition, and energy status of dairy goats. Eighteen multiparous Toggenburg goats were randomly assigned to dietary treatments in a crossover experimental design (14 d treatment periods separated by a 7 d washout interval): 30 g/d of calcium salts of fatty acids (Control) or 30 g/d of a rumen unprotected CLA supplement containing 29.9% of trans-10, cis-12 CLA as methyl esters (CLA). Lipid supplements were mixed into a concentrate and fed individually to animals 3 times a day as a total mixed ration component. The DMI, milk yield, milk protein and lactose content and secretion, and somatic cell count were unaffected by CLA treatment. On the other hand, milk fat content and yield were reduced by 19.9 and 17.9% in CLA-fed goats. Reduced milk fat yield in CLA-fed goats was a consequence of a lower secretion of both preformed and de novo synthesized fatty acids. The CLA treatment also changed the milk fatty acid profile, which included a reduction in the concentration of SFA (2.5%), increased MUFA and PUFA (5.6 and 5.4%, respectively), and a pronounced increase (1576%) in milk fat trans-10, cis-12 CLA. Consistent with the high milk fat trans-10, cis-12 CLA content, all desaturase indexes were reduced in milk fat from CLA-fed goats. The MFD induced by CLA reduced the energy required for milk production by 22%, which was accompanied by an improvement in the estimated energy balance (P < 0.001), greater blood glucose concentration (P < 0.05), and a trend for increased BW (P = 0.08). Approximately 7.2% of trans-10, cis-12 CLA was estimated to escape from rumen biohydrogenation and indirect comparisons with data obtained from other studies suggest equivalent MFD between dietary CLA in the methyl ester form and rumen protected sources. Thus, despite the apparent low degree of rumen protection, our results suggest that methyl esters of CLA could be an alternative to rumen protected CLA supplements due to manufacturing and cost advantages.

Corn silage and high-moisture corn grain are commonly recognized as risk factors for biohydrogenation-induced milk fat depression and may be due to the high concentration of linoleic acid (C18:2) in corn. Corn silage and corn grain have a low concentration of fatty acids (FA), but due to their high inclusion rate in diets they contribute substantially to unsaturated FA intake. The first objective of this study was to characterize the contribution of individual plant parts to total FA in whole-plant chopped corn. The second objective was to characterize the variation in FA profile in commercial silage and grain hybrids and evaluate the relationship between FA profile and other nutrients. To determine the location of FA in the corn plant, 4 stalks from 4 different commercial hybrids were separated into stalk, husk and shank, leaves, cob, and kernels. On a dry matter basis, 80.5% of total FA were in the kernels, 11.8% in the leaves, 5.1% in the stalk, 1.7% in the cob, and 1.0% in the husk and shank. More than 96% of the oleic acid (C18:1) and 92.5% of the C18:2 was in the kernels, whereas 71.0% of the linolenic acid (C18:3) was in the leaves. Next, the FA composition of fresh whole-plant chopped corn from 124 silage hybrids and grain from 72 grain hybrids was determined over 2 yr from test plots in Pennsylvania. Last, to extend the characterization, FA composition of whole-plant corn silage from 45 hybrids grown in test plots in South Dakota were characterized. In the fresh whole-plant chopped corn from PA test plots, C18:2 as a percentage of total FA averaged from 48.7% in 2013 (percentiles: 10th = 45.2, 90th = 52.2) and 48.0% in 2014 (percentiles: 10th = 44.1, 90th = 49.4). Concentration of C18:2 in corn grain averaged 57.5% in the 2013 (percentiles: 10th = 53.4, 90th = 60.8) and 56.1% in 2014 (percentiles: 10th = 53.5, 90th = 59.4). In the corn silage from South Dakota, the concentration of C18:2 as percentage of total FA averaged 45.4% (percentiles: 10th = 39.4, 90th = 50.2) and C18:2 concentration as a percent of dry matter averaged 1.1% (percentiles: 10th = 0.76, 90th = 1.41). An increase in the concentration of C18:2 was associated with a decrease in C18:3 in fresh whole-plant chopped corn and with a decrease in C18:1 in corn grain. Total FA and C18:2 (as a percentage of dry matter) were positively correlated with starch and negatively correlated with neutral detergent fiber in both fresh whole-plant chopped corn and corn silage samples, whereas no correlation with these traits was observed for C18:2 as a percentage of total FA. In conclusion, FA concentration and profile of corn silage reflects to a great extent the FA composition of kernels and the proportion of grain in the silage. The variation in C18:2 across hybrids provides the opportunity to develop selection programs to decrease C18:2 in corn silage and grain. Selection based on C18:2 concentration as a percent of total FA is preferred as this trait did not correlate with other nutritional properties.

2-Hydroxy-4-(methylthio)butanoate (HMTBa) is a methionine analog that has been observed to attenuate biohydrogenation (BH)-induced milk fat depression (MFD), possibly through reducing the shift to altered BH pathways. It has also been suggested that HMTBa increases microbial protein synthesis in the rumen. Our objectives were to stimulate BH-induced MFD and (1) verify HMTBa inhibition of BH-induced MFD and changes in milk fatty acids (FA) associated with altered rumen BH (i.e., trans-10 C18:1); and (2) determine the effect of HMTBa on milk fat (i.e., odd-and branchedchain FA) and urine biomarkers related to microbial N flow. Twenty-four multiparous cows (45.6 ± 8.5 kg of milk/d; mean ± standard deviation) and 12 primiparous cows (32.8 ± 3.1 kg of milk/d) were arranged in a crossover design. Treatments were unsupplemented control and HMTBa fed at 0.1% of diet dry matter intake. The experiment was 80 d and included a 10-d pretrial covariate period. Each experimental period included 2 phases that differed in risk for BH-induced MFD, including a 28-d low-risk phase (31.6% neutral detergent fiber, 21.8% starch, and no oil) and a 7-d moderate-risk phase (28.7% neutral detergent fiber, 28.1% starch, and 1.0% soybean oil). We found no interaction of treatment and parity. Milk fat yield (1.43 ± 0.51 kg/d) and milk fat trans-10 C18:1 (0.42 ± 0.08 g/100 g of FA) did not differ between treatments during the low-risk phase. However, during the moderate-risk phase, HMTBa maintained higher milk fat concentration (3.91 vs. 3.79%), tended to maintain higher milk fat yield (1.44 vs. 1.38 kg/d), and decreased milk fat trans-10 C18:1 (0.61 vs. 0.93% FA) compared with control. Additionally, HMTBa increased milk fat concentration and secretion of odd-and branched-chain FA by 5.3 and 10.2%, respectively, but urinary biomarkers of microbial N flow (i.e., purine derivatives) did not differ between treatments. However, rumen bacterial samples were not available to provide cow-or treatment-specific microbial protein-to-marker ratios, which is a critical source of variation. Additionally, transfer of odd-and branched-chain FA to milk is dependent on several factors that may affect interpretation of these biomarkers. In conclusion, HMTBa decreased absorption of alternate BH intermediates and maintained higher milk fat when feeding a diet with moderate-risk for MFD.

Dairy farm decision support systems (DSS) are tools which help dairy farmers to solve complex problems by improving the decision-making processes. In this paper, we are interested in newer generation, integrated DSS (IDSS), which additionally and concurrently: (1) receive continuous data feed from on-farm and off-farm data collection systems and (2) integrate more than one data stream to produce insightful outcomes. The scientific community and the allied dairy community have not been successful in developing, disseminating, and promoting a sustained adoption of IDSS. Thus, this paper identifies barriers to adoption as well as factors that would promote the sustained adoption of IDSS. The main barriers to adoption discussed include perceived lack of a good value proposition, complexities of practical application, and ease of use; and IDSS challenges related to data collection, data standards, data integration, and data shareability. Success in the sustainable adoption of IDSS depends on solving these problems and also addressing intrinsic issues related to the development, maintenance, and functioning of IDSS. There is a need for coordinated action by all the main stakeholders in the dairy sector to realize the potential benefits of IDSS, including all important players in the dairy industry production and distribution chain.