Feeding barley grain steeped in lactic acid modulates rumen fermentation patterns and increases milk fat content in dairy cows

The objectives of the present in vivo and in situ trials were to evaluate whether feeding barley grain steeped in lactic acid (LA) would affect rumen fermentation patterns, in situ dry matter (DM) degradation kinetics, and milk production and composition in lactating dairy cows. The in vivo trial involved 8 rumen-fistulated Holstein cows fed once daily a total mixed ration containing rolled barley grain (27% in DM) steeped for 48 h in an equal quantity of tap water (CTR) or in 0.5% LA (TRT) in a 2 × 2 crossover design. The in situ trials consisted of incubation of untreated rolled barley grain in cows fed CTR or TRT diets and of incubation of 3 different substrates including CTR or barley grain steeped in 0.5% or 1.0% LA (TRT1 and TRT2, respectively) up to 72 h in the rumen. Results of the in vivo trial indicated that cows fed the TRT diet had greater rumen pH during most intensive fermentation phases at 10 and 12 h post-feeding. The latter effect was associated with a shorter duration in which rumen pH was below 5.8 for cows fed the TRT diet (2.4 h) compared with CTR diet (3.9 h). Furthermore, cows fed the TRT diet had lower concentrations of volatile fatty acids at 2 and 4 h post-feeding. In addition, concentrations of preprandial volatile fatty acids were lower in the rumen fluid of cows fed the TRT diet. Results also showed that molar proportion of acetate was lower, whereas propionate tended to increase by feeding cows the TRT diet. Cows fed the TRT diet demonstrated greater rumen in situ lag time of substrate DM degradation and a tendency to lower the fractional degradation rate. Other in situ results indicated a quadratic effect of LA on the effective rumen degradability of substrates whereby the latter variable was decreased from CTR to TRT1 but increased for TRT2 substrate. Although the diet did not affect actual milk yield, fat-corrected milk, percentages of milk protein, and lactose and concentration of milk urea nitrogen, cows fed the TRT diet increased milk fat content and tended to increase fat:protein ratio in the milk. In conclusion, results demonstrated that treatment of barley grain with LA lowered the risk of subacute rumen acidosis and maintained high milk fat content in late-lactating Holstein cows fed diets based on barley grain.     

Temporal variation of milk fat globule diameter,fat and cholesterol content and milk epithelial cell gene expression in dairy cows

It was possible to show a connection between the temporal variation of milk fat globule diameter, fat and cholesterol content in milk and the expression of candidate genes in the mammary gland epithelial cells in milk. The beginning of lactation corresponded with higher levels of fat and cholesterol in the milk as a result of a higher expression of key enzymes in the purified bovine milk epithelial cells, paralleled with an increase in milk fat globule mean size.     

Relationships between rumen lipopolysaccharide and mediators of inflammatory response with milk fat production and efficiency in dairy cows

The main objective of this study was to evaluate correlative relationships between rumen lipopolysaccharide (LPS) and mediators of acute phase response with milk fat yield and efficiency in dairy cows challenged with graded amounts of barley grain in the diet. An additional aim of the study was to quantify the intercow variation in relation to milk fat production and acute phase response in cows fed graded amounts of grain. Eight primiparous, lactating Holstein cows (60 d in milk) were assigned to 1 of the 4 total mixed rations containing barley grain at 0, 15, 30, and 45% (dry matter basis) in a replicated 4 × 4 Latin square design. Free rumen LPS, plasma acute phase proteins, and milk fat content were quantified in multiple samples collected on d 5 and 7 of the measurement periods shortly before the morning feeding. Results showed markedly greater concentrations of rumen LPS with increasing dietary grain level. The correlative analysis revealed strong negative relationships between rumen LPS and milk fat content and yield. The predictor variable of rumen LPS explained 69% of the variation during the milk fat reduction of the cows. The stronger depression in milk fat percentage was obtained when rumen LPS exceeded a threshold of 5,564 ng/mL, corresponding to a milk fat content of 3.39%. The increase in concentration of rumen LPS was also associated with declines in milk fat yield and 3.5% fat-corrected milk (R² = 0.50), as well as milk energy efficiency (R² = 0.43). The correlative analysis also indicated that the increase of plasma C-reactive protein (CRP) in response to higher grain feeding was associated with a linear decrease of milk fat content and yield (R² = 0.28 to 0.46). Furthermore, the statistical analysis revealed high percentages of intercow variation related to milk fat variables, as well as the responses of rumen LPS and plasma CRP. Taken together, the current results implicate rumen LPS and the host CRP response in the lowering of milk fat content and milk energy efficiency in dairy cows fed high-grain diets. Further research is warranted to understand the mechanism(s) by which rumen LPS and inflammatory responses to LPS lower milk fat synthesis and milk energy efficiency and to develop novel strategies for their prevention.     

Alteration of rumen fermentation, milk fat synthesis, and nutrient utilization with mineral salts in dairy cows

The ability of mineral salts to alter rumen fermentation, rumen fluid dilution rate, milk fat synthesis, and nutrient utilization was investigated in dairy cows fed a high-concentrate, milk-fat depressing diet. Four rumen-fistulated Holstein cows were in a 4 X 4 Latin square design. Treatments consisted of: 1) basal (25% corn silage: 75% concentrate on a dry matter basis), 2) basal + 2.0% sodium chloride, 3) basal + 2.0% sodium bicarbonate, and 4) basal + 2.4% limestone. Addition of limestone to the basal diet reduced dry matter intake but increased efficiency of dietary nutrient utilization for milk synthesis. Sodium bicarbonate increased synthesis of milk fat. Sodium chloride also tended to increase milk fat synthesis whereas limestone had no effect. Milk yields (kg/day) and milk fat (%) for the four treatments were 1) 29.5, 2.40; 2) 29.3, 2.66; 3) 28.9, 3.26; and 4) 29.2, 2.32. Rumen fluid pH, dilution rate (%/hour), and molar percentage of acetate and propionate were: 1) 5.98, 10.3, 49 and 39; 2) 6.02, 12.4, 55 and 32; 3) 6.16, 12.2, 58 and 25; and 4) 5.92, 10.7, 51 and 38. Limestone was totally ineffective in altering ruminal pH, fluid dilution rate, molar percentages of acetate and propionate, and synthesis of milk fat. Improved feed efficiency for milk production after addition of limestone was related to an increase in starch digestion compared to the basal ration (95 versus 88%).     

Effect of dietary vitamin E on rumen biohydrogenation pathways and milk fat depression in dairy cows fed high-fat diets

Six lactating Holstein cows were assigned to a replicated Latin square design to test the effect of dietary vitamin E on milk fat depression and on the increased production of milk trans-10 C18:1 classically observed when feeding high doses of unsaturated fatty acids with low-fiber diets. Two diets (linseed diet and linseed diet + 12,000 IU of vitamin E/d) were compared during 2 periods of 21 d. The linseed diet presented a forage-to-concentrate ratio of 50:50 and contained extruded linseed (1.86 kg/d) and linseed oil (190 g/d). It was conceived to favor the “trans-11 to trans-10 shift” (low structural value and high level of unsaturated fatty acids). Milk yield and protein content were not affected by the diets. Milk of cows fed the linseed diet presented the typical symptoms of milk fat depression associated with a shift in biohydrogenation pathways: low fat content and high level of trans-10 C18:1. However, the high dose of dietary vitamin E provided significantly increased milk fat content (by 17.93%) and yield (by 15.56%) and decreased trans-10 C18:1 content (by 47.06%). In addition, it managed to significantly increase the daily yields of vaccenic (by 102.56%) and rumenic acids (by 56.67%). However, the sequence of administration of vitamin E influenced its effect, as vitamin E seemed to be more active in limiting the “trans-11 to trans-10 shift” when it was incorporated in the diet simultaneously with the fat. Once the shift had occurred, the subsequent addition of vitamin E was no longer able to completely counteract this process.     

Meta-analysis of rumination behavior and its relationship with milk and milk fat production,rumen pH,and total-tract digestibility in lactating dairy cows

Time spent ruminating is affected by diet and affects the rumen environment. The objective of the current study was to conduct a meta-regression to characterize the variation in rumination time and its relationship with milk and milk fat yields and variables mechanistically associated with milk fat synthesis, including rumen pH and total-tract digestibility. The analysis included 130 journal articles published between 1986 and 2018 that reported 479 treatment means from lactating Holsteins cows during established lactation. Milk yield averaged 34.3 kg/d (range 14.2–52.1 kg/d), milk fat averaged 3.47% (range 2.20–4.60%), and rumen pH averaged 6.1 (range 5.3–7.0). Rumination observation systems were categorized into 6 groups, but there was little difference in average rumination time among systems. The total time spent ruminating averaged 444 min/d (range 151–638 d) and occurred in 13.8 bouts/d (range 7.8–17.4 bouts/d) that averaged 32.7 min (range 20.0–48.1 min). Bivariate regressions were modeled to include the random effect of study, and correlations were evaluated through the partial R² that excluded variation accounted for by the random effect. Rumination time was quadratically increased with increasing milk fat yield (partial R² = 0.27) and milk fat percent (partial R² = 0.17). Rumination was also increased with increasing milk yield, dry matter intake, and rumen pH, and was quadratically related to dietary neutral detergent fiber (NDF) and total-tract NDF digestibility (partial R² = 0.10–0.27). Similar relationships were observed for rumination per unit of dry matter and NDF intake. The best-fit multivariate model predicting total rumination time included milk yield, milk fat yield, and concentration and accounted for 37% of the variation. Total-tract digestibility was available for 217 treatment means; when included in the model, the partial R² increased to 0.41. Last, principal component analysis was conducted to explore the relationship among variables. The first 2 principal components in the broad analyses explained 36.7% of the 39 variables evaluated, which included rumination bouts and time spent ruminating. In conclusion, rumination time was related to milk fat across a large number of studies, although it explained only a limited amount of the variation in milk fat.     

Effect of dietary fat in isocaloric rations on the nitrogen content of milk from Holstein cows

Effects of dietary fat on milk composition, particularly milk N, were evaluated using 12 lactating Holstein cows in a replicated 4 X 4 Latin-square design. Four complete mixed diets were formulated to contain either 0 or 3.5% added fat (grease) and either 1.6 or 1.7 Mcal NE1 (0, 1.6; 0, 1.7; 3.5, 1.6; and 3.5, 1.7). Diets were fed for ad libitum intake, and periods were 21 d. Adding fat to the diet depressed total milk N, casein N, and whey protein N concentration compared with no added fat at both energy densities. The proportion of total N in the casein fraction was not affected by dietary fat, whereas the proportion of N associated with the NPN fraction was increased with diets containing added fat. The high concentration diet (0, 1.7) contained the highest content of milk N and casein N. Added dietary fat increased the proportion of long-chain fatty acids (C18 and C18:1) in milk fat and decreased the proportion of short-chain fatty acids (C8 to C16). Daily milk yield was higher for the high energy fat diet (3.5, 1.7) than for the other three diets, which were similar. Blood concentrations of triglycerides, nonesterified fatty acids, and cholesterol were increased by diets containing added fat.     

Effect of 2-hydroxy-4-(methylthio)butanoate (HMTBa) on milk fat,rumen environment and biohydrogenation,and rumen protozoa in lactating cows fed diets with increased risk for milk fat depression

Biohydrogenation-induced milk fat depression (MFD) is a reduction in milk fat synthesis caused by bioactive fatty acids (FA) produced during altered ruminal microbial metabolism of unsaturated FA. The methionine analog 2-hydroxy-4-(methylthio)butanoate (HMTBa) has been shown to reduce the shift to the alternate biohydrogenation pathway and maintain higher milk fat yield in high-producing cows fed diets lower in fiber and higher in unsaturated FA. The objective of this experiment was to verify the effect of HMTBa on biohydrogenation-induced MFD and investigate associated changes in rumen environment and fermentation. Twenty-two rumen cannulated high-producing Holstein cows [168 ± 66 d in milk; 42 ± 7 kg of milk/d (mean ± standard deviation)] were used in a randomized design performed in 2 blocks (1 = 14 cows, 2 = 8 cows). Treatments were control (corn carrier) and HMTBa (0.1% of diet dry matter). The experiment included a 7-d covariate period followed by 3 phases that fed diets with increasing risk of MFD. The diet during the covariate and low-risk phase (7 d) was 32% neutral detergent fiber with no additional oil. The diet during the moderate-risk phase (17 d) was 29% neutral detergent fiber with 0.75% soybean oil. Soybean oil was increased to 1.5% for the last 4 d. The statistical model included the random effect of block and time course data were analyzed with repeated measures including the random effect of cow and tested the interaction of treatment and time. There was no effect of block or interaction of block and treatment or time. There was no overall effect of treatment or treatment by time interaction for dry matter intake, milk yield, and milk protein concentration and yield. Overall, HMTBa increased milk fat percent (3.2 vs. 3.6%) and yield (1,342 vs. 1,543 g/d) and there was no interaction of treatment and dietary phase. Additionally, HMTBa decreased the concentration of trans-10 18:1 in milk fat and rumen digesta. Average total ruminal concentration of volatile FA across the day and total-tract dry matter and fiber digestibility were not affected by HMTBa, but HMTBa increased average rumen butyrate and decreased propionate concentration and increased total protozoa abundance. Additionally, HMTBa increased the fractional rate of α-linoleic acid clearance from the rumen following a bolus predominantly driven by a difference in the first 30 min. Plasma insulin was decreased by HMTBa. In conclusion, HMTBa prevented the increase in trans FA in milk fat associated with MFD through a mechanism that is independent of total volatile FA concentration, but involves modification of rumen biohydrogenation. Decreased propionate and increased butyrate and ruminal protozoa may also have functional roles in the mechanism.     

Changes in rumen fermentation and bacterial community in lactating dairy cows with subacute rumen acidosis following rumen content transplantation

Rumen microbiota intervention has long been used to cure ruminal indigestion in production and has recently become a research hotspot. However, how it controls the remodeling of rumen bacterial homeostasis and the restoration of rumen fermentation in cows of subacute ruminal acidosis (SARA) remains poorly understood. This study explored changes in rumen fermentation and bacterial communities in SARA cows following rumen content transplantation (RCT). The entire experiment comprised 2 periods: the SARA induction period and the RCT period. During the SARA induction period, 12 ruminally cannulated lactating Holstein cows were selected and allocated into 2 groups at random, fed either a conventional diet [CON; n = 4; 40% concentrate, dry matter (DM) basis] or a high-grain diet (HG; n = 8; 60% concentrate, DM basis). After the SARA induction period, the RCT period began. The HG cows were randomly divided into 2 groups: the donor-recipient (DR) group and the self-recipient (SR) group, and their rumen contents were removed completely before RCT. For the DR group, cows received 70% rumen content from the CON cows, paired based on comparable body weight; for the SR group, each cow received 70% rumen content, self-derived. The results showed that nearly all rumen fermentation parameters returned to the normal levels that the cows had before SARA induction after 6 d of transplantation, regardless of RCT. The concentrations of acetate, valerate, and total volatile fatty acids (VFA) were not recovered in the SR cows, whereas all of them were recovered in the DR cows. The amplicon sequencing results indicated that both the SR and DR cows rebuild their rumen bacterial homeostasis quickly within 4 d after RCT, and the DR group showed a higher level of bacterial community diversity. At the genus level, the DR cows displayed an improved proportion of unclassified Ruminococcaceae and Saccharofermentans compared with the SR cows. Correlation analysis between the rumen bacteria and rumen fermentation suggested some potential relationships between the predominant transplantation-sensitive operational taxonomic units and VFA. Co-occurrence network analysis revealed that RCT affected only those rumen bacterial taxa that showed weak interactions with other taxa and did not affect the pivotal rumen bacteria with high levels of co-occurrence. Our findings indicate that RCT contributes to the restoration of rumen bacterial homeostasis and rumen fermentation in cows suffering from SARA without affecting the core microbiome.     

Densitometric determination of the fat content of milk and milk products

An automatable method for determining the fat content of milk and milk products was developed using a precise densitometer based on the principle of the resonant frequency U-tube oscillation. The sample is first decomposed with hydrochloric acid and the fat extracted with hexane. The density of the fat/hexane solution obtained varies with the fat content of the sample. Milk, cream and cheese samples were analysed. The mean values from the densitometric determination of the fat content did not differ from the mean values obtained by the corresponding gravimetric reference methods.     

Dietary fat and nitrogen composition of milk from lactating cows

Effects of dietary fat on milk composition, particularly milk N, were evaluated in lactating dairy cows at two stages of lactation. Complete mixed diets containing 0, 3.5, or 7% of the diet DM as animal fat were fed to 12 cows in a 3 X 3 Latin square. Cows were divided into two status categories based on stage of lactation resulting in two squares of early and two of late lactation cows. Percentages of fat, solids, lactose, and protein were decreased and ash increased in the milk of cows fed the 7% fat diet. Percent of casein N was elevated while nonprotein N was depressed by fat fed in diet. Percentage of solids and protein was higher and lactose lower for cows in late lactation than for those in early lactation. Dietary fat reduced the proportion of short-chain fatty acids in milk fat and increased C18:0 and C18:1 fatty acids. Digestibility of DM, energy, and fiber was not significantly affected by dietary fat, but estimates decreased with increased dietary fat. Cows in late lactation had a higher digestibility of dietary fractions than early lactation cows even though intakes of DM were similar.     

Assessment of rumen bacteria in dairy cows with varied milk protein yield

The present study was conducted to assess rumen bacteria in lactating cows with different milk protein yield, aiming to understand the role of rumen bacteria in this trait. Cows with high milk protein yield (high milk yield and high milk protein content, HH; n = 20) and low milk protein yield (low milk yield and low milk protein content, LL; n = 20) were selected from 374 mid-lactation Holstein dairy cows fed a high-grain diet. Measurement of the rumen fermentation products showed that the concentrations of ruminal total volatile fatty acids, propionate, butyrate, and valerate and the proportion of isobutyrate were higher in the HH cows than in the LL cows. Amplicon sequencing analysis of the rumen bacterial community revealed that the richness (Chao 1 index) of rumen microbiota was higher in the LL cows than in the HH cows. Among the 10 predominant bacterial phyla (relative abundance being >0.10%, present in >60% of animals within each group), the relative abundance of Proteobacteria was 1.36-fold higher in the HH cows than in the LL cows. At the genus level, the relative abundance of Succinivibrio was significantly higher and that of Clostridium tended to be higher in the LL cows than in the HH cows. Sharpea was 2.28-fold enriched in the HH cows compared with the LL cows. Different relationships between the relative abundances of rumen microbial taxa and volatile fatty acid concentrations were observed in the HH and the LL animals, respectively. Succinivibrio and Prevotella were positively correlated with acetate, propionate, and valerate in the LL cows, whereas Sharpea was positively correlated with propionate and valerate concentrations in the HH cows. Collectively, our results revealed that rumen bacterial richness and the relative abundances of several bacterial taxa significantly differed between dairy cows with high and low milk protein yields, suggesting the potential roles of rumen microbiota contributing to milk protein yield in dairy cows.