Effect of feeding increasing levels of whole cottonseed on milk and milk components, milk fatty acid profile, and total-tract digestibility in lactating dairy cows.

Dietary fat is fed to increase energy intake and provide fatty acids (FA) to support milk fat production. Oilseeds contain unsaturated FA that increase the risk for biohydrogenation-induced milk fat depression, but FA in whole cottonseed (WCS) are expected to be slowly released in the rumen and thus have a lower risk for biohydrogenation-induced milk fat depression. Our hypothesis was that increasing dietary WCS would increase milk fat yield by providing additional dietary FA without induction of milk fat depression. Four primiparous and 8 multiparous lactating Holstein cows, 136 ± 35 and 127 ± 4 DIM, respectively, were arranged in a replicated 4 × 4 Latin square design with 21-d periods. Treatments were WCS provided at 0%, 3.4%, 6.8%, and 9.9% of dietary dry matter, and WCS was substituted for cottonseed hulls and soybean meal to maintain dietary fiber and protein. Treatment did not change milk yield. There was a treatment-by-parity interaction for milk fat percent and yield with a quadratic decreased in primiparous cows but no effect of WCS in multiparous cows. Cottonseed linearly increased milk fat trans-10 18:1 in primiparous cows but not in multiparous cows. Increasing WCS increased milk preformed (18C) FA yield and partially overcame the trans-10 18:1 inhibition of de novo FA synthesis in the primiparous cows. Apparent transfer of 18C FA from feed to milk decreased in all cows as WCS increased, but the magnitude of the change was greater in primiparous cows. Increasing WCS decreased total-tract apparent dry matter, organic matter, and neutral detergent fiber digestibility. There was no change in total FA digestibility. However, 18C FA digestibility tended to be decreased in both parities and 16C FA digestibility was quadratically increased in multiparous cows but not changed in primiparous cows. Total fecal flow of intact WCS increased as WCS level increased, but fecal flow of intact seeds as a percentage consumed was similar across treatments. Fecal flow of intact seeds was greater in multiparous cows (4.3% vs. 1.1% of consumed). Plasma concentrations of glucose, nonesterified FA, triglycerides, and insulin were not changed. However, plasma urea-N increased with increasing WCS. Plasma gossypol increased with WCS (0.08-1.15 µg/mL) but was well below expected toxic levels. In conclusion, WCS maintained milk and milk component yield when fed at up to 9.9% of the diet to multiparous cows without concerns of gossypol toxicity, but primiparous cows were more susceptible to biohydrogenation-induced milk fat depression in the current trial. This highlights the interactions of parity with diet composition when feeding rumen-available unsaturated fat to dairy cows.

A review of recent studies on the enrichment of eggs and poultry meat with omega-3 polyunsaturated fatty acids: novel findings and unanswered questions.

Studies from our laboratory over the past decade have yielded new information with regard to the dietary enrichment of eggs and poultry meat with omega-3 (n-3) polyunsaturated fatty acids (PUFA) but have also generated a number of unanswered questions. In this review, we summarize the novel findings from this work, identify knowledge gaps, and offer possible explanations for some perplexing observations. Specifically discussed are: 1) Why feeding laying hens and broilers an oil rich in stearidonic acid (SDA; 18:4 n-3), which theoretically bypasses the putative rate-limiting step in the hepatic n-3 PUFA biosynthetic pathway, does not enrich egg yolks and tissues with very long-chain (VLC; ≥20 C) n-3 PUFA to the same degree as obtained by feeding birds oils rich in preformed VLC n-3 PUFA; 2) Why in hens fed an SDA-rich oil, SDA fails to accumulate in egg yolk but is readily incorporated into adipose tissue; 3) How oils rich in oleic acid (OA; 18:1 n-9), when co-fed with various sources of n-3 PUFA, attenuates egg and tissue n-3 PUFA contents or rescues egg production when co-fed with a level of docosahexaenoic acid (DHA; 22:6 n-3) that causes severe hypotriglyceridemia; and 4) Why the efficiency of VLC n-3 PUFA deposition into eggs and poultry meat is inversely related to the dietary content of α-linolenic acid (ALA; 18:3 n-3), SDA, or DHA.

Direct Comparison of 18 Carbon n-3 and n-6 Fatty Acids at Equal Levels in an Oil Blend on Tissue Enrichment of Long-Chain Polyunsaturated Fatty Acid in Broiler Chickens.

BACKGROUND: Omega-3 (n-3) and omega-6 (n-6) polyunsaturated fatty acids (PUFAs) are of interest because of their health effects. However, most experiments use natural oils and are confounded by PUFA concentrations and other fatty acids (FAs) that impact biosynthesis of the very long-chain derivatives (VLC). OBJECTIVES: To directly compare the effect of 18 C n-3 or n-6 FA fed at similar rates on their elongation and desaturation to VLC PUFA and their incorporation into tissues. METHODS: Oil blends that substituted ∼23% points of stearidonic acid (SDA) with alpha-linolenic acid (ALA), gamma-linolenic acid (GLA), or linoleic acid (LA) while minimizing differences in other FA were prepared. COBB500 broilers were fed the oil blends at 1.25% of the diet from day 14-35 age. RESULTS: There was greater enrichment of VLC PUFA in breast, thigh, liver, and plasma when diets were supplemented with high-SDA and high-GLA oil blends than high-ALA and high-LA oil blends. The efficiency of VLCn-3 PUFA synthesis from SDA and ALA was lower than the efficiency of VLCn-6 PUFA synthesis from GLA and LA, suggesting that the elongation and desaturation enzymes more efficiently utilized n-6 substrates. The efficiency of biotransformation of SDA to VLCn-3 PUFA was greater than that of high-ALA, and synthesis of VLCn-6 PUFA from GLA was higher than that of high-LA in breast, thigh, liver, and plasma. There were minimal effects on tissue-saturated and monounsaturated FA. CONCLUSIONS: The high-SDA and high-GLA oil blends efficiently enriched tissues with their VLC-PUFA more than high-ALA and high-LA treatments.

Effects of the timing of protein infusion on the daily rhythms of milk synthesis and plasma hormones and metabolites in dairy cows.

Milk synthesis exhibits a daily rhythm that is modified by the timing of feed intake. However, it is unknown how specific nutrients entrain this daily rhythm. Amino acids have an important role in milk synthesis, and may have a role in entrainment of mammary circadian rhythms. The objective of this study was to determine the effects of intestinally absorbed protein on daily rhythms of milk and milk component synthesis and key plasma hormones and metabolites. Nine lactating Holstein cows were assigned to 1 of 3 treatment sequences in a 3 × 3 Latin square. Treatments included abomasal infusions of 500 g/d of sodium caseinate either continuously throughout the day (CON), for 8 h/d from 0900 to 1700 h (DAY), or for 8 h/d from 2100 to 0500 h (NGT). Cows were milked every 6 h during the final 8 d of each period. A 24-h rhythm was fit to data using cosine analysis and the amplitude and acrophase were determined. Night infusion of protein decreased the daily milk yield and milk protein yield by 8.2% and 9.2%, respectively. Milk fat yield was increased 5.5% by DAY and milk fat concentration was increased 8.8% by NGT. Milk yield exhibited a daily rhythm in all treatments, with NGT increasing the amplitude of the daily rhythm 33% compared with CON. Milk fat concentration fit a daily rhythm in CON and NGT, but not DAY, whereas milk protein concentration fit a daily rhythm in CON and DAY, but not NGT. Moreover, DAY abolished the daily rhythm of plasma glucose concentration, but induced rhythms of plasma insulin and nonesterified fatty acid concentrations. Results suggest that feeding increased protein levels during the early part of the day may increase milk fat yield and modify energy metabolism through increased daily variation in insulin-stimulated lipid release, but additional research focused on feeding multiple diets across the day is required.

Kinetics of omega-3 fatty acid transfer to milk differs between fatty acids and stage of lactation in dairy cows.

Fatty acids (FA) differ in their transfer efficiencies and metabolic partitioning and lactating cows provide a robust model to investigate kinetics of FA transport. The objective was to compare kinetics of n-3 polyunsaturated FA (PUFA) trafficking through plasma and into milk. In the first experiment, ten ruminally cannulated multiparous Holstein cows were used in a crossover design with 7 d periods. Cows were milked at 6 h intervals and abomasal treatments provided a single dose of 80.1 g of α-linolenic acid as free FA (ALA-FFA) or 45.5 g EPA and 32.9 g DHA (LCn3-FFA). Transfer of n-3 PUFA to milk was nearly 50% higher for ALA-FFA than LCn3-FFA (48.2 and 32.7% of the bolus) and fit a bi-exponential model. Rapid transport of n-3 PUFA, assumed to be directly through chylomicrons, was nearly twice as high in ALA-FFA than LCn3-FFA and the subsequent slow transport, assumed to be indirect transfer through tissue recycling, was over 2.5-fold higher in LCn3-FFA than in ALA-FFA. Plasma analysis revealed LCn3-FFA enriched phospholipids and cholesterol esters, which had a slow clearance. In the second experiment, 4 cows received a bolus of a mixture of ALA, EPA, and DHA prepartum while not lactating and around d 10, 55, and 225 of lactation. Transfer of ALA to milk did not differ between stages of lactation, but DHA was lower in early compared to mid and late lactation. In conclusion, dietary ALA is rapidly and efficiently transferred to milk in cows while EPA and DHA are rapidly incorporated into plasma or tissue fractions not available to the mammary gland. This demonstrates clear differences in trafficking and partitioning of n-3 PUFA that ultimately impact tissue and organelle enrichment with implications for effective doses.

Comparison of Ahiflower oil containing stearidonic acid to a high-alpha-linolenic acid flaxseed oil at two dietary levels on omega-3 enrichment of egg yolk and tissues in laying hens.

Enrichment of egg yolks with very long chain omega-3 fatty acids (VLCn-3 FA) is of interest because of their beneficial effects on human health. The ability of Ahiflower® oil (AHI; Buglossoides arvensis), which is naturally rich in stearidonic acid (SDA), and a high-alpha-linolenic acid (ALA) flaxseed (FLAX) oil to enrich eggs and tissues of laying hens with VLCn-3 FA was investigated. Forty 54-week-old Hy-Line W-36 White Leghorn hens were fed a diet that contained soybean oil (control; CON) or AHI or FLAX oils at 7.5 or 22.5 g/kg of the diet in substitution for the soybean oil for 28 days. Dietary treatments had no effects on egg number or components or follicle development. Total VLCn-3 FA contents of egg yolk, liver, breast, thigh, and adipose tissue were greater in the n-3 treatments compared to CON, with the greatest increase observed at the higher oil level, especially for AHI oil which had the greater VLCn-3 enrichment than FLAX in yolk (p < 0.001). Efficiency of VLCn-3 enrichment of egg yolks was decreased with n-3 oils and by increasing oil level with lowest efficiency at 22.5 g/kg FLAX. In conclusion, both SDA-rich (AHI) and ALA-rich (FLAX) oils increased VLCn-3 FA deposition into egg yolks and hens' tissues, but dietary AHI oil promoted a greater enrichment than comparative amounts of FLAX oil, especially in liver and egg yolks.

Feeding laying hens docosa hexaenoic acid-rich microalgae oil at 40 g/kg diet causes hypotriglyceridemia, depresses egg production, and attenuates expression of key genes affecting hepatic triglyceride synthesis and secretion, but is rescued by dietary co-supplementation of high-oleic sunflower oil.

The primary goal of this study was to investigate the effect of feeding White Leghorn hens graded levels of a docosahexaenoic acid (DHA)-rich microalgae oil (MAO) on productive performance and enrichment of eggs with very long-chain (VLC) omega-3 (n-3) polyunsaturated fatty acids (PUFA). Forty-nine-week-old hens (8 per diet) were fed the following diets for 28 d: 1) A corn-soybean meal-based diet with no supplemental oil (CON); 2) CON + 10 g/kg MAO; 3) CON + 20 g/kg MAO; 4) CON + 30 g/kg MAO; 5) CON + 40 g/kg MAO; 6) CON + 40 g/kg MAO + 20 g/kg high-oleic sunflower oil (HOSO); and 7) CON + 40 g/kg MAO + 40 g/kg HOSO. Diets 6 and 7 were included because we previously reported that co-feeding high-oleic acid oils with n-3 PUFA-containing oils attenuated egg yolk n-3 PUFA contents vs. feeding hens the n-3 oils alone. All data were collected on an individual hen basis. Egg VLC n-3 PUFA enrichment plateaued, in terms of statistical significance, at the 30 g/kg MAO level (266 mg/yolk). Hens fed 40 g/kg MAO had greatly attenuated measures of hen performance, marked liver enlargement, an altered ovarian follicle hierarchy, greatly lowered circulating triglyceride levels, and depressed hepatic expression of key genes involved in triglyceride synthesis and secretion. As compared to hens fed 40 g/kg MAO alone, feeding hens 40 g/kg MAO co-supplemented with HOSO (Diets 6 and 7) restored egg production, ovarian morphology, and all other measures of hen productive performance to CON levels, elevated plasma triglyceride levels, prevented liver enlargement, and increased the hepatic expression of key genes involved in triglyceride synthesis and secretion. In conclusion, MAO can greatly enrich hens' eggs with VLC n-3 PUFA, but its recommended dietary inclusion should not exceed 20 g/kg. This would allow for near-maximal yolk VLC n-3 PUFA enrichment without impairing hen productive performance, altering the ovarian follicle hierarchy or, based on the work of others, presumably imparting off-flavors in the egg.

Fatty Acid Transfer from Blood to Milk Is Disrupted in Mothers with Low Milk Production, Obesity, and Inflammation.

BACKGROUND: Obesity is associated with chronic inflammation and is a risk factor for insufficient milk production. Inflammation-mediated suppression of LPL could inhibit mammary uptake of long-chain fatty acids (LCFAs; >16 carbons). OBJECTIVES: In an ancillary case-control analysis, we investigated whether women with low milk production despite regular breast emptying have elevated inflammation and disrupted transfer of LCFAs from plasma into milk. METHODS: Data and specimens from a low milk supply study and an exclusively breastfeeding control group were analyzed, with milk production measured by 24-h test-weighing at 2-10 wk postpartum. Low milk supply groups were defined as very low (VL; <300 mL/d; n = 23) or moderate (MOD; ≥300 mL/d; n = 20) milk production, and compared with controls (≥699 mL/d; n = 18). Serum and milk fatty acids (weight% of total) were measured by GC, serum and milk TNF-α by ELISA, and serum high-sensitivity C-reactive protein (hsCRP) by clinical analyzer. Group differences were assessed by linear regression models, chi-square exact tests, and Kruskal-Wallis nonparametric tests. RESULTS: VL cases, as compared with MOD cases and controls, had higher prevalence of elevated serum hsCRP (>5 mg/L; 57%, 15%, and 22%, respectively; P = 0.004), detectable milk TNF-α (67%, 32%, and 33%, respectively; P = 0.04), and obesity (78%, 40%, and 22%, respectively; P = 0.003). VL cases had lower mean ± SD LCFAs in milk (60% ± 3%) than MOD cases (65% ± 4%) and controls (66% ± 5%) (P < 0.001). Milk and serum LCFAs were strongly correlated in controls (r = 0.82, P < 0.001), but not in the MOD (r = 0.25, P = 0.30) or VL (r = 0.20, P = 0.41) groups (Pint < 0.001). CONCLUSIONS: Mothers with very low milk production have significantly higher obesity and inflammatory biomarkers, lower LCFAs in milk, and disrupted association between plasma and milk LCFAs. These data support the hypothesis that inflammation disrupts normal mammary gland fatty acid uptake. Further research should address impacts of inflammation and obesity on mammary fatty acid uptake for milk production.

Validation of an alternate method for monitoring the presence of cows at the feed bunk in a Calan Broadbent Feeding System using a 3-axis, data-logging accelerometer.

Daily dry matter intake is a key observation in dairy nutrition, and observation of feeding behavior provides insight into the physiological control of hunger and satiety that regulate intake. The objective of the study was to develop and validate an alternative method to observe feeding behavior, including meal length and frequency, in a Calan Broadbent Feeding System (American Calan) using a 3-axis accelerometer (Hobo Pendant G, Onset Computer Corp.). Sensors were mounted between the door and the feed divider using commonly available materials without making permanent modifications to the feeding system. Forty-eight sensors were deployed with a recording frequency of 30 s for the last 7 d of each period in a crossover experiment with 24 multiparous and 24 primiparous animals housed in a freestall barn. The tilt angle on the Z-axis was used to determine when the door was open to indicate feeding activity. The sensor system was in very high agreement with 6 h of visual observation (Cohen's κ = 0.92 ± 0.014; estimate ± 95% confidence interval). The minimum intermeal interval is the time between 2 feeding bouts that is still considered one meal. This essential criterion to characterize meals was calculated by determining the intersection of a mixture of Gaussian distributions fitted to the log-transformed between-feeding intervals. The best fitting mixture of Gaussian distributions was determined with the distribution module of JMP Pro 14.3.0 (SAS Institute Inc.). The minimum intermeal interval was 31.3 min using the best fitting model, a mixture of 3 Gaussian distributions. Using the determined minimum intermeal interval, meal length averaged 37.3 min/meal and meal frequency averaged 7.3 meals/d. In conclusion, data-logging 3-axis accelerometers are adequate to monitor presence of cows in the feed gate in the Calan Broadbent Feeding System, and this approach allows for reasonable estimation of meal length and frequency.

TRB3 Deletion Has a Limited Effect on Milk Fat Synthesis and Milk Fat Depression in C57BL/6N Mice.

BACKGROUND: Regulation of the endoplasmic reticulum (ER) stress pathway is critical to mammary epithelial cell function throughout pregnancy, lactation, and involution. Treatment with trans-10, cis-12 conjugated linoleic acid (t10c12CLA) suppresses mammary lipogenesis and stimulates the ER stress pathway. The ER stress pathway includes tribbles pseudokinase 3 (TRB3), a protein that regulates cellular energy and insulin signaling. OBJECTIVES: Our objective was to describe the effect of TRB3 deficiency on milk fat synthesis and determine if TRB3 deficiency protects against suppression of mammary lipogenesis. METHODS: First, mammary Trb3 expression was observed throughout pregnancy and lactation using ancillary microarray data (n = 4/time point). Second, intake, litter growth, and milk clot fatty acid (FA) profile of Trb3 knockout (KO) C57BL/6N mice were compared with wild-type (WT) and heterozygous (HET) mice throughout first (n ≥ 8/group) and second (n ≥ 6/group) lactation. Lastly, the interaction between Trb3 genotype and 2 treatments that suppress mammary lipogenesis, t10c12CLA and high safflower oil (HO) diet, was investigated in a 2 × 2 factorial design (n ≥ 6/group). RESULTS: Trb3 expression was higher during late pregnancy and lactation. Trb3 KO and HET mice had lower feed intake, dam weight, and litter growth throughout first, but not second, lactation than WT mice. Treatment with t10c12CLA decreased litter growth (28%; P < 0.0001) and feed intake (8%; P < 0.0001) regardless of Trb3 genotype. When fed the HO diet, Trb3 KO mice had 17% higher mammary de novo synthesized FAs (<16 carbons; P int = 0.002) than WT mice. Mammary ER stress and lipogenic genes were mostly unaltered by Trb3 deficiency. CONCLUSIONS: Overall, TRB3 plays a minor role in regulating mammary lipogenesis, because Trb3 deficiency had only a limited protective effect against diet-induced suppression of lipogenesis.

Comparison of Ahiflower oil containing stearidonic acid to a high-alpha-linolenic acid flaxseed oil at two levels on tissue omega-3 enrichment in broilers.

Enrichment of broiler meat with very long-chain omega-3 fatty acids (VLCn-3 FA) is of interest because of their beneficial effects on human health. The ability of Ahiflower® (AHI) oil (Buglossoides arvensis), which naturally contains stearidonic acid (SDA), and a high-alpha-linolenic acid (ALA) flaxseed (FLAX) oil to enrich VLCn-3 FA contents in broilers tissues was investigated. Fifty-five Cobb 500 chicks were fed from days 12 to 35 of life either a control (CON) diet that contained 27.9 g/kg soybean oil or AHI or FLAX oils, each individually at 7.5 or 22.5 g/kg of the diet in substitution for soybean oil (all on an as fed basis). Total VLCn-3 FA contents were greater in breast, thigh, liver, adipose tissue, and plasma of all n-3 treatments compared to CON, with the greatest increase observed at the highest level of AHI and FLAX oils (p < 0.001). AHI oil at 7.5 g/kg promoted the most efficient synthesis and deposition of VLCn-3 in broiler tissues measured as deposition of VLCn-3 FA in tissues relative to intake of n3 FA. In conclusion, both ALA and SDA oils increased VLCn-3 FA deposition in tissues, but there were diminishing returns when increasing dietary levels of the oils.

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.

Supplemental dietary oils rich in oleic acid or linoleic acid attenuate egg yolk and tissue n-3 polyunsaturated fatty acid contents in laying hens co-fed oils enriched in either stearidonic acid or α-linolenic acid.

We previously reported that when laying hens were fed diets supplemented with oils enriched in α-linolenic acid (ALA) and oleic acid (OA), the deposition of n-3 PUFA in egg yolk was attenuated as compared to feeding hens a diet supplemented with the ALA-rich oil alone. The present work extends those findings to another n-3 PUFA-rich oil (stearidonic acid [SDA]-enriched soybean oil) and two other high-OA oils, suggesting that the effect is not plant oil-specific. Feeding hens a supplemental linoleic acid (LA)-rich oil plus an oil rich in either SDA or ALA also attenuated egg yolk ALA and SDA contents (Experiment 1), or egg yolk and liver ALA contents (Experiment 2), respectively, as compared to feeding the SDA- or ALA-rich oils alone. Future work should focus on the lack of neutrality of OA and LA in relation to n-3 PUFA nutrition.

Dietary SFAs and ω-6 Fatty Acids Alter Incorporation of ω-3 Fatty Acids into Milk Fat of Lactating CD-1 Mice and Tissues of Offspring.

BACKGROUND: Methods to increase the amount of omega-3 (n-3) PUFAs in milk are desirable for neonatal health. The n-3 PUFA, α-linolenic acid (18:3n-3), can be elongated to EPA (20:5n-3) and DHA (22:6n-3). n-6 PUFAs suppress tissue n-3 PUFA incorporation, but the effect of SFAs is not clear. OBJECTIVES: In this study, we compared the effects of SFAs and n-6 PUFAs on n-3 PUFA incorporation into milk and tissues of lactating mice and tissues of their offspring. METHODS: Female CD-1 mice were bred at 8 wk of age. All experimental diets included 3% flaxseed oil and were begun on day 8 of lactation: low-fat diet (LFD); high-SFA diet (SAT), with an additional 12% saturated oil; or high-linoleic-acid diet (HLA), with 12% high-linoleic-acid oil (% kcal, carbohydrates:fat:protein: LFD, 49:24:27; both SAT and HLA, 35:46:19; n = 5/treatment). After 5 d, pup stomach milk clot FA profiles, tissue FA profiles in dams and pups, and mammary and hepatic expression of lipid metabolism genes in dams were analyzed. Data were analyzed by ANOVA with treatment diet as a fixed effect. RESULTS: Dams in all groups had similar total milk fat concentrations, but both SAT and HLA decreased the concentration of n-3 PUFAs (SAT: -23%; HLA: -31%) compared with LFD, and HLA increased milk n-6 FAs by 347% compared with SAT. SAT pups had n-3 PUFA tissue concentrations similar to LFD, but HLA pups had lower n-3 PUFAs than SAT pups in multiple tissues (liver, -32%; kidney, -29%; heart, -28%; muscle, -18%). Mammary expression of lipid metabolism genes was mostly unchanged, but hepatic expression of elongases and desaturases was decreased with SAT compared with LFD [elongation of very-long-chain fatty acid (Elov)5, -42%; Elov6, -64%; fatty acid desaturase (Fads)1, -33%; Fads2, -44%]. CONCLUSIONS: HLA decreased n-3 PUFA concentrations across multiple pup tissues compared with SAT. This suggests that high dietary n-6 PUFAs suppress n-3 PUFA incorporation in neonates.

Physical characterization of fatty acid supplements with varying enrichments of palmitic and stearic acid by differential scanning calorimetry.

Saturated fatty acid supplements commonly fed to dairy cows differ in their fatty acid (FA) profile. Some supplements with very high enrichments of palmitic acid (PA) or stearic acid (SA) have been reported to have low total-tract digestibility. Saturated FA have the potential to form crystalline structures at high purity that may affect digestibility. Differential scanning calorimetry (DSC) is a thermal technique commonly used in materials science to measure the change in heat flow as energy is absorbed or released from a sample during heating, and it was used to characterize a series of experimental and commercial fat supplements. Our hypothesis was that products with very high enrichment of either PA or SA would differ in thermal characteristics compared with those that include moderate levels of a second FA because of the formation of secondary crystalline structures, which may contribute to decreased digestibility. First, replicated runs demonstrated low variation in melting temperature (MT) and enthalpy (coefficient of variation <4%). The effect of physical form was evaluated by comparing an initial thermal cycle to a second, successive thermal cycle after samples had resolidified in the test pan. Melting temperature was slightly increased by 1.3°C by the second cycle compared with the first, but there was no change in enthalpy. Next, supplements with 98% SA, 98% PA, and an SA/PA (44%/55%) blend with undetectable levels of unsaturated FA were compared. Melting temperature of the SA/PA mixture was 61.2°C and similar to the expected MT of PA (62.9°C). However, the MT of the high-purity SA and PA were increased to 73.7°C and 67.8°C, respectively, and enthalpy increased by 12.5% compared with the SA/PA blend. An FA stock highly enriched in SA (>98%) had the highest MT, and one moderately enriched in PA (∼85%) that contained 10.1% unsaturated FA had the lowest enthalpy value of all FA supplements and experimental stocks that were characterized. Differential scanning calorimetry may be useful to screen and design supplements with improved physical properties that may be associated with digestibility.

Short communication: Diet digestibility measured from fecal samples and associations with phenotypic and genetic merit for milk yield and composition.

Selection for improved feed utilization is of high interest globally but is limited by the high cost of obtaining feed intake for individual cows and relies on indirect measures of feed efficiency. Supplementing selection with mechanistic measures of feed use could make selection for feed utilization more direct and effective. The objectives of this study were to evaluate fecal sampling as a method of determining digestive efficiency of individual cows and to evaluate associations of digestive efficiency with genetic and phenotypic merit for milk yield and composition. Fecal samples were obtained manually from the rectum of 90 Holstein cows in the morning, afternoon, and evening on a single date and composited across the day. The fecal samples were dried, ground, and stored. Diet and fecal neutral detergent fiber (NDF) were determined using the filter bag method, and indigestible NDF was determined in situ with a 12-d rumen incubation. Fecal NDF (60.1%) and indigestible NDF (41.9%) were higher than that from feed samples (14.2 and 35.9%, respectively). Total-tract digestibility was calculated using the marker ratio method. Total-tract dry matter (DM) digestibility averaged 66.0 ± 2.4% and total-tract NDF digestibility averaged 42.8 ± 3.0%. Higher milk fat percent and genetic merit for milk fat percent were associated with greater NDF and DM digestibility. Milk yield was negatively associated with NDF and DM digestibility. Fecal sampling is a feasible method to directly measure digestive efficiency, and substantial variation was observed among cows. Given significant between-cow variation and associations with milk fat percent and genetic merit for milk fat percent, potential selection for total-tract NDF digestibility estimated via fecal sampling warrants further exploration.

Dietary Fat Does Not Overcome trans-10, cis-12 Conjugated Linoleic Acid Inhibition of Milk Fat Synthesis in Lactating mice.

Trans-10, cis-12 conjugated linoleic acid (CLA) is a potent inhibitor of milk fat synthesis in the cow and similarly reduces milk fat in rodents. The objective of this study was to determine whether dietary fat can overcome CLA inhibition of milk fat concentration in lactating mice. Wild type C57Bl/6J mice (n = 31) were fed semipurified diets containing either low fat (LF; 4% fat) or high fat (HF; 23.6% fat) starting 4-6 days postpartum. Dietary fat was increased by inclusion of high oleic sunflower oil. After 2 days on the experimental diets, lactating dams were orally dosed with either water (control) or trans-10, cis-12 CLA (20 mg/day) for 5 days. CLA treatment decreased pup growth similarly in both HF and LF diets. Milk fat percent was increased over 16% by the HF diet and decreased over 12% by CLA, but there was no interaction of dietary fat and CLA. Both CLA and the HF diet reduced the proportion of short- and medium-chain fatty acids that originate from de novo synthesis, and there was no interaction of diet and CLA. CLA had no effect on the percent of preformed fatty acids, but the HF diet increased their abundance. Dietary fat and CLA both modified mammary expression of lipogenic enzymes and regulators, but no interactions were observed. In conclusion, CLA reduced milk fat concentration and litter growth, but these effects were not overcome by increased dietary fat from high oleic sunflower oil. CLA inhibition of milk fat in the mammary gland is not substrate dependent, and the mechanism is independent from dietary supply of oleic acid.

Night-restricted feeding of dairy cows modifies daily rhythms of feed intake, milk synthesis and plasma metabolites compared with day-restricted feeding.

The timing of feed intake can alter circadian rhythms of peripheral tissues. Milk synthesis displays a daily rhythm across several species, but the effect of feeding time on these rhythms is poorly characterised. The objective of this experiment was to determine if the time of feed intake modifies the daily patterns of milk synthesis, plasma metabolites and body temperature in dairy cows. Sixteen lactating Holstein dairy cows were randomly assigned to one of the two treatment sequences in a cross-over design with 17 d periods. Treatments included day-restricted feeding (DRF; feed available from 07.00 to 23.00 hours) and night-restricted feeding (NRF; feed available from 19.00 to 11.00 hours). Cows were milked every 6 h on the last 7 d of each period, and blood samples were collected to represent every 4 h over the day. Peak milk yield was shifted from morning in DRF to evening in NRF, while milk fat, protein and lactose concentration peaked in the evening in DRF and the morning in NRF. Plasma glucose, insulin, NEFA and urea nitrogen concentration fit daily rhythms in all treatments. Night feeding increased the amplitude of glucose, insulin and NEFA rhythms and shifted the daily rhythms by 8 to 12 h (P < 0·05). Night feeding also phase-delayed the rhythm of core body temperature and DRF increased its amplitude. Altering the time of feed availability shifts the daily rhythms of milk synthesis and plasma hormone and metabolite concentrations and body temperature, suggesting that these rhythms may be entrained by food intake.

Kinetics of trans-10, cis-12-conjugated linoleic acid transfer to plasma and milk following an abomasal bolus in lactating dairy cows.

Trans-10, cis-12-conjugated linoleic acid (CLA) is a potent bioactive fatty acids (FA) that causes milk fat depression in lactating animals. FA are transferred to milk directly through chylomicrons and indirectly by recycling through other tissues. The objective of this study was to characterise the kinetics of trans-10, cis-12 CLA transfer to plasma and milk after a single bolus infusion. Five multiparous mid-lactation cows received a single abomasal bolus infusion of an enriched CLA mixture providing 15 g of trans-10, cis-12 CLA and 15 g of cis-9, trans-11 CLA over a 30-min period. Plasma concentration of trans-10, cis-12 and cis-9, trans-11 CLA peaked 2 h post-bolus, reaching 0·29 and 0·38 % of total plasma FA, respectively, and returned to pre-bolus values at 72 h post-infusion. Milk trans-10, cis-12 CLA yield and concentration peaked 14 h post-bolus (0·25 g/h) and was not detectable in milk after 86 h. Total apparent transfer of trans-10, cis-12 CLA to milk was 41 %, with 73 % transferred to milk through the direct pool (chylomicrons) and the remaining 27 % transferred through the indirect pool (tissue recycling). Compartmental modelling revealed the existence of a transient unavailable pool of trans-10, cis-12 CLA in extravascular tissues represented primarily by the mammary gland, which slowly exchanges with an available pool for secretion in milk fat and transfer to milk. In conclusion, trans-10, cis-12 CLA is predominantly transferred to milk through the direct pathway; however, how this CLA isomer is processed within the mammary gland requires further investigation.