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.

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.

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.

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.

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.

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.

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.

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.

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.

Time-dependent effect of trans-10,cis-12 conjugated linoleic acid on gene expression of lipogenic enzymes and regulators in mammary tissue of dairy cows.

Trans-10,cis-12 conjugated linoleic acid (CLA) has been identified as an intermediate of rumen fatty acid biohydrogenation that caused milk fat depression (MFD) in the dairy cow. Previous studies in cows experiencing CLA- and diet-induced MFD have identified reduced mammary expression of the master lipogenic regulator sterol response element transcription factor 1 (SREBF1) and many of its dependent genes. To distinguish between primary mechanisms regulating milk fat synthesis and secondary adaptations to the reduction in milk fat, we conducted a time-course experiment. Eleven dairy cows received by abomasal infusion an initial priming dose of 6.25 g of CLA followed by 12.5 g/d delivered in multiple pulses per day for 5 d. Cows were milked 3×/d and mammary biopsies were obtained under basal condition (prebolus control) and 12, 30, and 120 h relative to initiation of CLA infusion. Milk fat concentration and yield decreased progressively reaching a nadir at 69 h (1.82% and 38.2 g/h) and averaged 2.03 ± 0.19% and 42.1 ± 4.10 g/h on the last day of treatment (±standard deviation). Expression of fatty acid synthase (FASN) and lipoprotein lipase (LPL) were decreased at 30 and 120 h compared with control. Expression of SREBF1 and THRSP were also decreased at 30 and 120 h compared with control. Additionally, we failed to observe changes in other factors, including peroxisome proliferator-activated receptor γ and liver × receptor ß and milk fat globular membrane proteins, during CLA treatment. However, expression of milk fat globular membrane proteins were decreased after 14 d of diet-induced MFD in samples from a previous experiment, indicating a possible long-term response. The rapid decrease in lipogenic enzymes, SREBF1, and THRSP provide strong support for their transcriptional regulation as a primary mechanism of milk fat depression.

Acetate Dose-Dependently Stimulates Milk Fat Synthesis in Lactating Dairy Cows.

Background: Acetate is a short-chain fatty acid (FA) that is especially important to cows because it is the major substrate for de novo FA synthesis. However, the effect of acetate supply on mammary lipid synthesis is not clear.Objective: The objective of this experiment was to determine the effect of increasing acetate supply on milk fat synthesis in lactating dairy cows.Methods: Six multiparous lactating Holstein cows were randomly assigned to treatments in a replicated design to investigate the effect of acetate supply on milk fat synthesis. Treatments were 0 (control), 5, 10, and 15 mol acetate/d continuously infused into the rumen for 4 d. Rumen short-chain FAs, plasma hormones and metabolites, milk fat concentration, and milk FA profile were analyzed on day 4 of each treatment. Polynomial contrasts were used to test the linear and quadratic effects of increasing acetate supply.Results: Acetate increased milk fat yield quadratically (P < 0.01) by 7%, 16%, and 14% and increased milk fat concentration linearly (P < 0.001) by 6%, 9%, and 11% for 5, 10, and 15 mol acetate/d, respectively, compared with the control treatment. Increased milk fat yield predominantly was due to a linear increase in 16-carbon FAs (P < 0.001) and a quadratic increase in de novo synthesized FAs (<16-carbon FAs; P < 0.01), indicating that there was stimulation of de novo synthesis pathways. Apparent transfer of acetate to milk fat was 33.4%, 36.2%, and 20.6% for 5, 10, and 15 mol/d, respectively. Acetate infusion linearly increased the relative concentration of rumen acetate (P < 0.001) before feeding, but not after feeding. Acetate linearly increased plasma ß-hydroxybutyric acid by 29%, 50%, and 78%, respectively, after feeding compared with the control treatment (P < 0.01).Conclusions: Increasing acetate supply to lactating cows increases milk fat synthesis, suggesting that nutritional strategies that increase ruminal acetate absorption would be expected to increase milk fat by increasing de novo FA synthesis.

Deficiency of stearoyl-CoA desaturase-1 aggravates colitogenic potential of adoptively transferred effector T cells.

Stearoyl-CoA desaturase-1 (SCD1) is a lipogenic enzyme involved in the de novo biosynthesis of oleate (C18:1, n9), a major fatty acid in the phospholipids of lipid bilayers of cell membranes. Accordingly, Scd1KO mice display substantially reduced oleate in cell membranes. An altered SCD1 level was observed during intestinal inflammation; however, its role in modulating inflammatory bowel disease remains elusive. Herein, we investigated the colitogenic capacity of Scd1KO effector T cells by employing the adoptive T-cell transfer colitis model. Splenic effector T cells (CD4+CD25-) from age- and sex-matched wild-type (WT) and Scd1KO mice were isolated by FACS and intraperitoneally administered to Rag1KO mice, which were monitored for the development of colitis. At day 60 postcell transfer, Rag1KO mice that received Scd1KO CD4+CD25- T cells displayed accelerated and exacerbated colitis than mice receiving WT CD4+CD25- T cells. Intriguingly, Scd1KO CD4+CD25- T cells display augmented inflammatory cytokine profile and cellular membrane fluidity with a concomitant increase in proinflammatory saturated fatty acids, which we postulate to potentially underlie their augmented colitogenic potential.

Trans-10, cis-12 CLA dose-dependently inhibits milk fat synthesis without disruption of lactation in C57BL/6J mice.

BACKGROUND: Trans-10, cis-12 conjugated linoleic acid (10,12 CLA) is a potent inhibitor of milk fat synthesis in mammals. In the cow, 10 g/d of 10,12 CLA specifically and reversibly inhibits mammary lipogenesis, whereas substantially higher doses are not specific and cause a generalized inhibition of milk synthesis. OBJECTIVE: The objective of this study was to validate a lactating mouse model by establishing the dose response, specificity, and reversibility of the inhibition of milk fat synthesis by 10,12 CLA. METHODS: Lactating mice (C57BL/6J) received daily doses of 0 (control), 7, 20, or 60 mg of 10,12 CLA for 5 d during established lactation. A second group of lactating mice was treated with 20 mg/d of 10,12 CLA for 4 d and followed post-treatment to evaluate reversibility. RESULTS: CLA decreased pup growth with a 49% decrease occurring with 60 mg/d of CLA. Milk fat percentage was decreased 11% and 20% with the 7 and 20 mg/d dose, respectively, and all CLA treatments had a decreased concentration of de novo synthesized fatty acids (FAs) in milk fat. In agreement, 20 mg/d of 10,12 CLA decreased the lipogenic capacity of mammary tissue by 30% and mammary expression of FA synthase (Fasn), sterol response element binding protein 1 (Srebf1), and thyroid hormone responsive spot 14 (Thrsp) by 30-60%, whereas milk protein percentage and mammary expression of α-lactalbumin (Lalba) were unaltered. This dose of CLA reduced pup growth by nearly 20% and milk de novo synthesized FAs by >35%, and these effects were completely reversed 5 d after 10,12 CLA treatment was terminated. CONCLUSION: Inhibition of mammary lipogenesis by 10,12 CLA is dose-dependent in the mouse, with a specific and reversible reduction in milk fat synthesis at the 20 mg/d dose and additional nonspecific effects on milk synthesis at higher CLA doses.

Supplementation with fish oil as a source of n-3 fatty acids does not downregulate mammary lipogenesis in lactating mice.

The very long chain n-3 (ω-3) polyunsaturated fatty acids (VLCn3PUFAs) are potent regulators of hepatic lipid synthesis, but their effect on lipid synthesis in the lactating mammary gland is less well investigated. The objective of the present study was to examine effects of fish oil (FO) supplementation on mammary lipogenesis and the expression of lipogenic genes in mammary and hepatic tissues of lactating mice. Beginning on day 6 of lactation and continuing for 7 d, female C57BL/6J mice (n = 8/diet) were fed 1 of 3 dietary treatments: a 5%-fat diet containing mainly saturated fatty acids (FAs) (low-fat control) or 2 10%-fat diets, 1 enriched with FO as a source of VLCn3PUFAs and the other enriched with a safflower/palm oil mixture (high-fat control) as a source of oleic acid. Mammary lipogenic capacity, measured by (14)C-glucose incorporation into FAs by mammary explants, was similar among treatments, and there were no treatment effects on the proportion of de novo synthesized FAs in milk fat or on litter weight gain, a proxy for milk energy secretion. Also, there were no treatment effects on mammary mRNA abundance for key lipogenic enzymes and proteins involved in the regulation of milk lipid synthesis. In contrast, there was a treatment effect on hepatic lipogenesis, with FO resulting in a decrease of ~50% in hepatic lipid content and a similar downregulation of lipogenic gene expression compared with the 2 control diets. Overall, there were tissue-specific differences in dietary VLCn3PUFA effects on lipid synthesis with no observed effects for mammary lipogenic variables but marked reductions occurring in hepatic lipogenesis.

Induction of and recovery from milk fat depression occurs progressively in dairy cows switched between diets that differ in fiber and oil concentration.

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.

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.

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.