Milk fat depression (MFD) caused by intermediates of ruminal biohydrogenation commonly occurs in dairy cattle. The time course of recovery from MFD is important to mechanistic investigation and management of the condition. Nine cows were used in a repeated design, allowing analysis of recovery from diet-induced MFD. A high-fiber, low-oil diet was fed during the control and recovery periods, and a low-fiber, high-oil (LFHO) diet was fed during the induction period. Milk yield was not affected by treatment. Milk fat percentage and yield decreased progressively during induction and were lower by d 3 and 5, respectively. Milk fat concentration and yield increased progressively when cows were fed the recovery diet and were not different from control on d 19 and 15, respectively. Yield of de novo synthesized fatty acids (FA) decreased progressively during the induction period and was lower than that of controls by d 5. A biphasic response was seen for milk fat trans isomers, where trans-11 C18:1 and cis-9,trans-11 conjugated linoleic acid (CLA) were elevated initially and trans-10 C18:1 and trans-10,cis-12 CLA increased progressively during the induction period. A similar biphasic response was seen during recovery from MFD, with trans-10 C18:1 and trans-10,cis-12 rapidly decreasing initially and trans-11 C18:1 and cis-9,trans-11 CLA increasing slightly above control levels during the second phase. Recovery from diet-induced MFD occurs gradually with a short lag when dietary fiber and oil concentrations are corrected. This time course provides a framework to identify factors causing MFD and set expectations during recovery from MFD.

Effects of feeding a dry glycerin product (minimal 65% of food grade glycerol, dry powder) to 39 multiparous Holstein dairy cows (19 control and 20 glycerin-supplemented; lactation number = 2.2 +/- 1.3 SD) on feed intake, milk yield and composition, and blood metabolic profiles were investigated. Dry glycerin was fed at 250 g/d as a top dressing (corresponding to 162.5 g of glycerol/d) to the common lactating total mixed ration from parturition to 21 d postpartum. Individual milk was sampled from 2 consecutive milkings weekly and analyzed for components. Blood was sampled from the coccygeal vein at 4, 7, 14, and 21 (+/-0.92, pooled SD) d in milk and analyzed for urea nitrogen, glucose, insulin, nonesterified fatty acids, and beta-hydroxybutyrate. Urine was tested for the acetoacetate level weekly by using Ketostix. Average feed intake, milk yield and components, blood metabolites, and serum insulin concentrations were not affected by dry glycerin supplementation. Glycerin-supplemented cows experienced a more positive energy status (higher concentrations of plasma glucose, lower concentrations of plasma beta-hydroxybutyrate, and lower concentrations of urine ketones), which was observed during the second week of lactation, suggesting that energy availability may have been improved. This glucogenic effect of dry glycerin did not result in an increase in feed intake or milk yield during the first 3 wk of lactation, likely because of the relatively less negative energy status of cows transitioning into lactation. The tendency toward higher milk yield for glycerin-supplemented cows during wk 6 of lactation (52 vs. 46 kg/d) after the supplementation period (dry glycerin was terminated at wk 3 of lactation) suggested a potential benefit of dry glycerin on subsequent milk production, perhaps through changes in metabolism, which requires further investigation.

The effect of a high-palmitic acid fat supplement was tested in 12 high-producing (mean = 42.1 kg/d) and 12 low-producing (mean = 28.9 kg/d) cows arranged in a replicated 3 × 3 Latin square design. Experimental periods were 21 d, with 18d of diet adaptation and 3 d of sample collection. Treatments were (1) control (no supplemental fat), (2) high-palmitic acid (PA) supplement (84% C16:0), and (3) Ca salts of palm fatty acid (FA) supplement (Ca-FA). The PA supplement had no effect on milk production, but decreased dry matter intake by 7 and 9% relative to the control in high- and low-producing cows, respectively, and increased feed efficiency by 8.5% in high-producing cows compared with the control. Milk fat concentration and yield were not affected by PA relative to the control in high- or low-producing cows, although PA increased the yield of milk 16-C FA by more than 85 g/d relative to the control. The Ca-FA decreased milk fat concentration compared with PA in high-, but not in low-producing cows. In agreement, Ca-FA dramatically increased milk fat concentration of trans-10 C18:1 and trans-10, cis-12 conjugated linoleic acid (>300%) compared with PA in high-producing cows, but not in low-producing cows. No effect of treatment on milk protein concentration or yield was detected. The PA supplement also increased 16-C FA apparent digestibility by over 10% and increased total FA digestibility compared with the control in high- and low-producing cows. During short-term feeding, palmitic acid supplementation did not increase milk or milk fat yield; however, it was efficiently absorbed, increased feed efficiency, and increased milk 16-C FA yield, while minimizing alterations in ruminal biohydrogenation commonly observed for other unsaturated fat supplements. Longer-term experiments will be necessary to determine the effects on energy balance and changes in body reserves.

Ten ruminally cannulated Holstein cows were used in a crossover design that investigated changes in ruminal bacterial populations in response to induction and recovery from diet-induced milk fat depression (MFD). Further, the effect on the ruminal microbiota of the cows with diet-induced milk fat depression inoculated with rumen contents from non-milk fat-depressed donor cows was evaluated. Milk fat depression was induced during the first 10 d of each period by feeding a low-fiber, high-starch, and high-polyunsaturated fatty acid diet (26.1% neutral detergent fiber, 28.1% starch, 5.8% total fatty acids, and 1.9% C18:2), resulting in a 30% decrease in milk fat yield. Induction was followed by a recovery phase, where all cows were switched to a high-fiber, low-starch, and low-polyunsaturated fatty acid diet (31.8% neutral detergent fiber, 23% starch, 4.2% total fatty acids, and 1.2% C18:2) and were allocated to (1) control (no inoculation) or (2) ruminal inoculation with donor cow digesta (8 kg/d for 6 d). Ruminal samples were collected at the end of induction (d 10) and during recovery (d 13, 16, and 28), separated to solid and liquid fractions, extracted for DNA, PCR- amplified for the V1-V2 region of the 16S rRNA gene, and analyzed for bacterial diversity. Results indicated that bacterial communities were different between fractions. In each fraction, differences were significant between the induction (d 10) and recovery (d 13, 16, and 28) periods; however, differences were less apparent with time during the recovery period. The MFD (d 10) was typified by a reduction in the relative sequence abundance of Bacteroidetes and an increase in the relative sequence abundance of Firmicutes and Actinobacteria across both fractions. At the genus level, relative sequence abundance of unclassified Lachnospiraceae, Butyrivibrio, Bulleidia, and Coriobacteriaceae were higher on d 10 and were positively correlated with trans-10,cis-12 CLA and the trans-10 isomer, suggesting their potential role in altered biohydrogenation reactions. A switch to the recovery diet resulted in a sharp increase in the Bacteroidetes lineages and a decrease in Firmicutes members on d 13; however, this shift appears to stabilize by d 28, indicating the restoration process for ruminal bacteria from an altered state is gradual and complex. Inoculation of 10% of rumen contents from non-MFD donor cows to MFD cows revealed this procedure had transient effects on only a few bacterial populations, and such effects disappeared after d 16 following cessation of inoculation. It can be concluded that alterations in milk FA profiles at induction are preceded by microbial alterations in the rumen driven by dietary changes.

The ruminant provides a powerful model for understanding the temporal dynamics of gastrointestinal microbial communities. Diet-induced milk fat depression (MFD) in the dairy cow is caused by rumen-derived bioactive fatty acids, and is commonly attributed to the changes in the microbial population. The aim of the present study was to determine the changes occurring in nine ruminal bacterial taxa with well-characterised functions, and abundance of total fungi, ciliate protozoa and bacteria during the induction of and recovery from MFD. Interactions between treatment and time were observed for ten of the twelve populations. The total number of both fungi and ciliate protozoa decreased rapidly (days 4 and 8, respectively) by more than 90 % during the induction period and increased during the recovery period. The abundance of Streptococcus bovis (amylolytic) peaked at 350 % of control levels on day 4 of induction and rapidly decreased during the recovery period. The abundance of Prevotella bryantii (amylolytic) decreased by 66 % from day 8 to 20 of the induction period and increased to the control levels on day 12 of the recovery period. The abundance of Megasphaera elsdenii and Selenomonas ruminantium (lactate-utilising bacteria) increased progressively until day 12 of induction (. 170 %) and decreased during the recovery period. The abundance of Fibrobacter succinogenes (fibrolytic) decreased by 97 % on day 4 of induction and increased progressively to an equal extent during the recovery period, although smaller changes were observed for other fibrolytic bacteria. The abundance of the Butyrivibrio fibrisolvens/Pseudobutyrivibrio group decreased progressively during the induction period and increased during the recovery period, whereas the abundance of Butyrivibrio hungatei was not affected by treatment. Responsive taxa were modified rapidly, with the majority of changes occurring within 8 d and their time course was similar to the time course of the induction of MFD, demonstrating a strong correlation between changes in ruminal microbial populations and MFD. Key words: Milk fat depression: Rumen microbes: Dairy cows: Conjugated linoleic acidDiet-induced milk fat depression (MFD) is caused by the inhibition of milk fat synthesis by bioactive fatty acids (FA) synthesised by rumen microbes, and is a well-studied example of the interaction between dietary nutrients, the gastrointestinal microbiome and tissue physiology. Rumen microbes perform a wide range of functions and are classically grouped within niches based on their predominant substrate or enzyme activity. Moreover, rumen microbes biohydrogenate unsaturated FA, resulting in the formation of trans isomers as intermediates. The rate, extent and pathways of biohydrogenation (BH) are commonly attributed to the microbial population present in the rumen. Previous investigations quantifying ruminal microbial populations have been conducted after diet adaptation periods; however, the time course of adaptation to a new diet is not well characterised.Importantly...

Milk fatty acid (FA) profile has been previously used as a predictor of enteric CH 4 output in dairy cows fed diets supplemented with plant oils, which can potentially impact ruminal fermentation. The objective of this study was to investigate the relationships between milk FA and enteric CH 4 emissions in lactating dairy cows fed different types of forages in the context of commonly fed diets. A total of 81 observations from three separate 3 × 3 Latin square design (32-day periods) experiments including a total of 27 lactating cows (96 ± 27 days in milk; mean ± SD) were used. Dietary forages were included at 60% of ration dry matter and were as follows: (1) 100% corn silage, (2) 100% alfalfa silage, (3) 100% barley silage, (4) 100% timothy silage, (5) 50 : 50 mix of corn and alfalfa silages, (6) 50 : 50 mix of barley and corn silages and (7) 50 : 50 mix of timothy and alfalfa silages. Enteric CH 4 output was measured using respiration chambers during 3 consecutive days. Milk was sampled during the last 7 days of each period and analyzed for components and FA profile. Test variables included dry matter intake (DMI; kg/day), NDF (%), ether extract (%), milk yield (kg/day), milk components (%) and individual milk FA (% of total FA). Candidate multivariate models were obtained using the Least Absolute Shrinkage and Selection Operator and Least-Angle Regression methods based on the Schwarz Bayesian Criterion. Data were then fitted into a random regression using the MIXED procedure including the random effects of cow, period and study. A positive correlation was observed between CH 4 and DMI (r = 0.59, P < 0.001), whereas negative associations were observed between CH 4 and cis9-17:1 (r = − 0.58, P < 0.001), and trans8, cis13-18:2 (r = − 0.51, P < 0.001). Three different candidate models were selected and the best fit candidate model predicted CH 4 with a coefficient of determination of 0.84 after correction for cow, period and study effects and was: CH 4 (g/day) = 319.7 − 57.4 × 15:0 − 13.8 × cis9-17:1 − 39.5 × trans10-18:1 − 59.9 × cis11-18:1 − 253.1 × trans8, cis12-18:2 − 642.7 × trans8, cis13-18:2 − 195.7 × trans11, cis15-18:2 + 16.5 × DMI. Overall and linear prediction biases of all models were not significant ( P > 0.19). Milk FA profile and DMI can be used to predict CH 4 emissions in dairy cows across a wide range of dietary forage sources.

Thirteen multiparous Holstein cows were used in a crossover design that tested the effect of lysolecithin in diets differing in neutral detergent fiber (NDF) and unsaturated fatty acid (FA) concentrations. Experimental periods were 20 d in length and included two 10-d phases. A standard fiber and lower fat diet was fed the first 10 d (30.5% NDF, no added oil, lower-risk phase) and a lower NDF and higher oil diet was fed during the second 10 d (29.0% NDF and 2% oil from whole soybeans and soybean oil, high-risk phase). Treatments were control and 10 g/d of lysolecithin (LYSO) extended in a ground corn carrier. Milk was sampled on d 0, 5, and 10 of each phase for determination of fat and protein concentration and FA profile. We found no effect of treatment or treatment by time interaction for dry matter intake, milk yield, or milk protein concentration. A treatment by time interaction was observed for milk fat concentration and yield. Milk fat concentration was higher in LYSO on d 5 of the lower-risk phase, but decreased progressively in both treatments during the high-risk phase. Milk fat yield was not different among treatments during the lower-risk phase, but was lower in LYSO on d 15 and tended to be lower on d 20 during the high-risk phase. Concentrations of milk de novo FA decreased and preformed FA increased during the high-risk phase, but we found no effect of treatment or treatment by time interactions. We noted an effect of time, but no treatment or treatment by time interactions for milk trans FA isomers. Briefly, trans-11 C18:1 and cis-9,trans-11 conjugated linoleic acid progressively decreased as trans-10 C18:1 and trans-10,cis-12 conjugated linoleic acid progressively increased during the high-risk phase. The LYSO increased milk fat concentration when feeding a higher fiber and lower unsaturated FA diet, but decreased milk fat yield when feeding a lower fiber and higher unsaturated FA diet, although biohydrogenation pathways and capacity did not appear to be modified. The effect of lysolecithin on rumen fermentation warrants further investigation, but is not recommended when feeding lower fiber and higher unsaturated fat diets.

Vitamin B 12 is synthesized by prokaryotes in the rumens of dairy cows-and this has implications in human nutrition since humans rely on consumption of dairy products for vitamin B 12 acquisition. However, the concentration of vitamin B 12 in milk is highly variable, and there is interest in determining what causes vitamin B 12 variability. We collected 92 temporally linked rumen, fecal, blood, and milk sample sets from Holstein cows at various stages of lactation fitted with rumen cannula and attempted to define which bacterial genera correlated well with vitamin B 12 abundance. The level of vitamin B 12 present in each sample was measured, and the bacterial population of each rumen, fecal, and milk sample (n ϭ 263) was analyzed by 16S rRNA-targeted amplicon sequencing of the V4 region. The bacterial populations present in the rumen, small intestine, and milk were highly dissimilar. Combined diet and lactation status had significant effects on the composition of the microbiota in the rumen and in the feces. A high ruminal concentration of vitamin B 12 was correlated with the increased abundance of Prevotella, while a low ruminal concentration of vitamin B 12 was correlated with increased abundance of Bacteroidetes, Ruminiclostridium, and Butyrivibrio. The ultimate concentration of vitamin B 12 is controlled by the complex interaction of several factors, including the composition of the microbiota. Bacterial consumption of vitamin B 12 in the rumen may be more important in determining overall levels than bacterial production. IMPORTANCE In this paper, we examined the microbiome of the bovine rumen, feces, and milk and attempted to understand how the bacterial communities at each site affected the production and movement of vitamin B 12 around the animal's body. It was determined that the composition of the bovine rumen microbiome correlates well with vitamin B 12 concentration, indicating that the rumen microbiota may be a good target for manipulation to improve production of this important vitamin.