Milk Fat Depression Research Articles

Review: Effect of essential fatty acids and conjugated linoleic acid on the adaptive physiology of dairy cows during the transition period.

Cows fed total mixed rations (silage-based) may not receive as much essential fatty acids (EFAs) and conjugated linoleic acids (CLAs) as cows fed pasture-based rations (fresh grass) containing rich sources of polyunsaturated fatty acids. CLA-induced milk fat depression allows dairy cows to conserve more metabolisable energy, thereby shortening the state of negative energy balance and reducing excessive fat mobilisation at early lactation. EFAs, particularly α-linolenic acid, exert anti-inflammatory and antioxidative properties, thereby modulating immune functions. Thus, combined EFA and CLA supplementation seems to be an effective nutritional strategy to relieve energy metabolism and to improve immune response, which are often compromised during the transition from late pregnancy to lactation in high-yielding dairy cows. There has been extensive research on this idea over the last two decades, and despite promising results, several interfering factors have led to varying findings, making it difficult to conclude whether and under what conditions EFA and CLA supplementations are beneficial for dairy cows during the transition period. This article reviews the latest studies on the effects of EFA and CLA supplementation, alone or in combination, on dairy cow metabolism and health during various stages around parturition. Our review article summarises and provides novel insights into the mechanisms by which EFA and/or CLA influence markers of metabolism, energy homeostasis and partitioning, immunity, and inflammation revealed by a deep molecular phenotyping.

Effect of oil and tannin supplementation on intake, milk yield and milk composition of dairy cows

Supplementation of oils rich in polyunsaturated fatty acids and condensed tannin has been known as a feeding approach to improve healthy fatty acids in ruminant milk, but it can cause an adverse effect on feed intake and animal performance. This study aimed to investigate the effect of feeding oil alone or in combination with grape seed tannin extract (GSTE) on feed intake, milk yield and composition of dairy cows. Sixteen low production dairy cows in mid-lactation fed a basal diet based on agro-industrial by-products were arranged to a completely randomized design for a 6-week duration. Animals were fed basal diet without oil and GSTE inclusion (CON), 2.5% DM soybean oil (SBO), 2.5% DM blend of soybean oil and tuna fish oil at 3:2 w:w (SFO), or SFO plus 0.4% DM GSTE (OCT). The results showed that DM intake was reduced (P < 0.05) by 14.4% in OCT relative to CON. Milk yield was not affected by oil and GSTE supplementation, but SFO and OCT strongly depressed milk fat, protein and total solids (P < 0.001). In conclusion, in a low production cow diet based on agro-industrial by-products containing high lipid, supplementation of oil and GSTE should be considered in the aspects of feed intake and milk composition.

Milk and plasma proteomes from cows facing diet-induced milk fat depression are related to immunity, lipid metabolism and inflammation

Milk proteins are a source of bioactive molecules for calves and humans that may also reflect the physiology and metabolism of dairy cows. Dietary lipid supplements are classically used to modulate the lipid content and composition of bovine milk, with potential impacts on the nutrient’s homeostasis and the systemic inflammation of cows that remains to be more explored. This study aimed at identifying discriminant proteins and their associated pathways in twelve Holstein cows (87 ± 7 days in milk), multiparous and non-pregnant, fed for 28 d a diet either, supplemented with 5% DM intake of corn oil and with 50% additional starch from wheat in the concentrate (COS, n = 6) chosen to induce a milk fat depression, or with 3% DM intake of hydrogenated palm oil (HPO, n = 6) known to increase milk fat content. Intake, milk yield and milk composition were measured. On d 27 of the experimental periods, milk and blood samples were collected and label-free quantitative proteomics was performed on proteins extracted from plasma, milk fat globule membrane (MFGM) and skimmed milk (SM). The proteomes from COS and HPO samples were composed of 98, 158 and 70 unique proteins, respectively, in plasma, MFGM and SM. Of these, the combination of a univariate and a multivariate partial least square discriminant analyses reveals that 15 proteins in plasma, 24 in MFGM and 14 in SM signed the differences between COS and HPO diets. The 15 plasma proteins were related to the immune system, acute-phase response, regulation of lipid transport and insulin sensitivity. The 24 MFGM proteins were related to the lipid biosynthetic process and secretion. The 14 SM proteins were linked mainly to immune response, inflammation and lipid transport. This study proposes discriminant milk and plasma proteomes, depending on diet-induced divergence in milk fat secretion, that are related to nutrient homeostasis, inflammation, immunity and lipid metabolism. The present results also suggest a higher state of inflammation with the COS diet.

Transcriptome Profile in Dairy Cows Resistant or Sensitive to Milk Fat Depression

Feeding linseed to dairy cows results in milk fat depression (MFD), but there is a wide range of sensitivity among cows. The objectives of this study were to identify target genes containing SNP that may play a key role in the regulation of milk fat synthesis in cows resistant or sensitive to MFD. Four cows were selected from a dairy farm after a switch from a control diet to a linseed-rich diet; two were resistant to MFD with a high milk fat content in the control (4.06%) and linseed-rich (3.90%) diets; and two were sensitive to MFD with the milk fat content decreasing after the change from the control (3.87%) to linseed-rich (2.52%) diets. Transcriptome and SNP discovery analyses were performed using RNA-sequencing technology. There was a large number of differentially expressed genes in the control (n = 1316) and linseed-rich (n = 1888) diets. Of these, 15 genes were detected as key gene regulators and harboring SNP in the linseed-rich diet. The selected genes MTOR, PDPK1, EREG, NOTCH1, ZNF217 and TGFB3 may form a network with a principal axis PI3K/Akt/MTOR/SREBP1 involved in milk fat synthesis and in the response to diets that induced MFD. These 15 genes are novel candidate genes to be involved in the resistance or sensitivity of dairy cows to milk fat depression.

Comparison of milk and grass composition from grazing Irish dairy herds with and without milk fat depression

BackgroundThis study investigated the factors relating to pasture chemical and fatty acid (FA) composition that influence the milk fat percentage of spring calving, grazing dairy cows. The relationship between milk fat percentage and FA composition of the milk in these herds was also investigated.ResultsMilk protein percentage, milk casein percentage and cheddar cheese yield were increased in milk from HMF herds. Cows from LMF herds did not have negatively altered milk processability including rennet coagulation time (RCT), pH and ethanol stability. Crude protein, NDF, ADF, ether extract and total FA content of pasture was not different between LMF and HMF herds. Milk fat concentration of conjugated linoleic acid (CLA) t10, c12 was not different between HMF and LMF herds. Pre-grazing herbage mass and pasture content of crude protein, neutral detergent fibre (NDF) and total FA were similar between HMF and LMF herds. Pasture offered to LMF herds had a higher concentration of monounsaturated fatty acids (MUFA). A strong negative relationship (r = -0.40) was evident between milk fat percentage and pasture crude protein content for MMF herds (3.31–3.94% milk fat).ConclusionsThis research reports improved milk protein percentage, milk casein percentage and cheddar cheese yield from HMF herds compared to LMF herds. Milk processability was not impacted by low milk fat percentage. Pasture NDF and total fatty acid content was similar in HMF herds and LMF herds. Milk fat percentage had a strong negative association (r = -0.40) with pasture crude protein content in MMF herds (MF 3.31–3.94%). Correlation values between pasture chemical and FA composition and milk fat percentage in LMF herds and HMF herds were low, indicating that diet is not the only causative factor for variation in milk fat of grazing dairy cows. Comparison of milk fatty acid composition from herds with and without milk fat depression suggests that there may be other fatty acids apart from CLA t10, c12 that contribute to the inhibition of milk fat synthesis during milk fat depression in grazing herds.

Nutritional Quality of Milk Fat from Cows Fed Full-Fat Corn Germ in Diets Containing Cactus Opuntia and Sugarcane Bagasse as Forage Sources

We evaluated the performance, milk composition, and milk fatty acid profile of cows fed diets composed of cactus cladodes (Opuntia stricta [Haw.] Haw), sugarcane bagasse and increasing levels of full-fat corn germ (FFCG). We hypothesized that ground corn can be effectively replaced by FFCG when cactus cladodes and sugarcane bagasse are used as forage sources. The cows were randomly distributed into two 5 × 5 Latin Squares and fed five diets in which ground corn was progressively replaced with full-fat corn germ (FFCG; 0%, 25%, 50%, 75%, or 100% of substitution). Adding FFCG to the diet increased milk production and milk fat content and reduced milk protein content. Overall, FFCG reduced the proportion of saturated FAs and increased mono- and polyunsaturated FAs in milk, including CLA isomers. In addition, activity indices of stearoyl-CoA desaturase were reduced by increasing levels of FFCG. We conclude that the substitution of corn for FFCG in diets based on cactus cladodes and sugarcane bagasse positively modifies the FA profile of milk and could add commercial value to milk products (e.g., CLA-enriched milk). In addition, the milk fat response indicates that the basal diet was favorable to the rumen environment, preventing the trans-10 shift commonly associated with milk fat depression.

Integrating data from spontaneous and induced trans-10 shift of ruminal biohydrogenation reveals discriminant bacterial community changes at the OTU level.

Microbial digestion is of key importance for ruminants, and disturbances can affect efficiency and quality of products for human consumers. Ruminal biohydrogenation of dietary unsaturated fatty acids leads to a wide variety of specific fatty acids. Some dietary conditions can affect the pathways of this transformation, leading to trans-10 fatty acids rather than the more usual trans-11 fatty acids, this change resulting in milk fat depression in dairy cows. We combined data from an induced and spontaneous trans-10 shift of ruminal biohydrogenation, providing new insight on bacterial changes at different taxonomic levels. A trans-10 shift was induced using dietary addition of concentrate and/or unsaturated fat, and the spontaneous milk fat depression was observed in a commercial dairy herd. Most changes of microbial community related to bacteria that are not known to be involved in the biohydrogenation process, suggesting that the trans-10 shift may represent the biochemical marker of a wide change of bacterial community. At OTU level, sparse discriminant analysis revealed strong associations between this change of biohydrogenation pathway and some taxa, especially three taxa belonging to [Eubacterium] coprostanoligenes group, Muribaculaceae and Lachnospiraceae NK3A20 group, that could both be microbial markers of this disturbance and candidates for studies relative to their ability to produce trans-10 fatty acids.

Susceptibility to milk fat depression in dairy sheep and goats: Individual variation in ruminal fermentation and biohydrogenation.

Small ruminants are susceptible to milk fat depression (MFD) induced by marine lipid supplementation. However, as observed in dairy cows, there is wide individual variation in the response to MFD-inducing diets, which may be due to individual differences in ruminal processes. Therefore, we compared the ruminal responses of goats and sheep with varying degrees of MFD extent to improve our understanding of this complex syndrome. Our specific aims were to attempt to elucidate whether pre-existing variations in ruminal fermentation and biohydrogenation determine a higher tolerance or susceptibility to MFD, and whether the severity of MFD depends exclusively on the response to the diet. The trial was conducted with 25 does and 23 ewes fed a basal diet without lipid supplementation for 3 wk (control period). Then, 2% fish oil (FO) was added to the same diet for 5 additional weeks (MFD period). Based on the extent of the elicited MFD (i.e., the percentage variation between milk fat concentrations recorded at the end of the control and MFD periods), the 5 most responsive (RESPON+) and the 5 least responsive (RESPON-) animals were selected within each species. On the last day of each period, ruminal fluid samples were collected to examine fermentation parameters and fatty acid profiles. In general, the individual degree of MFD in sheep and goats did not seem to be predetermined by traits related to ruminal fermentation and biohydrogenation, including fatty acids that may serve as biomarkers of microorganisms. Regarding differences in the response to FO, the results suggest no link between MFD susceptibility and concentration of biohydrogenation intermediates such as trans-10-containing C18, C20, and C22 metabolites. The explanation for individual responses based on a shortage of ruminal acetate and 18:0 for mammary uptake also seems to be dismissed, based on the lack of variation in these compounds between RESPON+ and RESPON-. However, the concentration of unsaturated fatty acids provided by FO (e.g., cis-9 16:1, cis-11 18:1, and 20:5n-3) was higher in the rumen of RESPON+ than RESPON- ewes and does. Thus, although further research is needed, the extent of biohydrogenation of these fatty acids might be associated with tolerance or susceptibility to MFD.

Determination of Fat Depression Levels in Cow Milk Obtained from Edirne and Tekirdag Provinces

In this study, it was aimed to determine fat depression levels in cow milk obtained from Edirne and Tekirdağ provinces in May. Research data on this subject were obtained from a private dairy factory in May. Milk fat depression is a condition detected as a result of the milk fat levels of dairy cattle falling below 3.20%. Milk fat depression is an undesirable situation because it causes problems in obtaining dairy products. The data of May were compared with the 3.2% value, which is accepted as the milk fat depression threshold, with the help of a one-sample T-test. In this study, total fat levels were determined as 2.93% for Edirne and 3.32% for Tekirdağ. When the total fat levels were compared with the reference value, depression was observed in the Edirne province. However, depression has not been found in Tekirdağ province. Accordingly, milk obtained from Edirne in May is undesirable in terms of dairy production. In the province of Tekirdağ, such negativity has not been observed. In Edirne province, regulations should be made on animal feeding to increase milk fat levels. Fat depression analysis should be done in milk obtained from different provinces. Thus, it is of great importance for the economy of the country and human health to take urgent measures for feeding and animal breeding for places with depression.

Effects of PUFA-Rich Dietary Strategies on Ruminants' Mammary Gland Gene Network: A Nutrigenomics Review.

Although the inclusion of polyunsaturated fatty acids (PUFAs) in ruminants' diets appears to be a well-documented strategy to enrich milk with PUFAs, several gene networks that regulate milk synthesis and mammary gland homeostasis could be impaired. The objective of this literature review is to assess the effects of nutritional strategies focused on enriching milk with PUFAs on gene networks regulating mammary gland function and lipogenesis, as well as the impact of feed additives and bioactive compounds with prominent antioxidant potential on immune-oxidative transcriptional profiling, as a part of mammary gland homeostasis and health. The findings support the conclusion that PUFAs' inclusion in ruminants' diets more strongly downregulate the stearoyl-CoA desaturase (SCD) gene compared to other key genes involved in de novo fatty acid synthesis in the mammary gland. Additionally, it was revealed that seed oils rich in linoleic and linolenic acids have no such strong impact on networks that regulate lipogenic homeostasis compared to marine oils rich in eicosapentaenoic and docosahexaenoic acids. Furthermore, ample evidence supports that cows and sheep are more prone to the suppression of lipogenesis pathways compared to goats under the impact of dietary marine PUFAs. On the other hand, the inclusion of feed additives and bioactive compounds with prominent antioxidant potential in ruminants' diets can strengthen mammary gland immune-oxidative status. Considering that PUFA's high propensity to oxidation can induce a cascade of pro-oxidant incidences, the simultaneous supplementation of antioxidant compounds and especially polyphenols may alleviate any side effects caused by PUFA overload in the mammary gland. In conclusion, future studies should deeply investigate the effects of PUFAs on mammary gland gene networks in an effort to holistically understand their impact on both milk fat depression syndrome and homeostatic disturbance.

Elucidating genes and gene networks linked to individual susceptibility to milk fat depression in dairy goats.

Dietary supplementation with marine lipids modulates ruminant milk composition toward a healthier fatty acid profile for consumers, but it also causes milk fat depression (MFD). Because the dairy goat industry is mainly oriented toward cheese manufacturing, MFD can elicit economic losses. There is large individual variation in animal susceptibility with goats more (RESPO+) or less (RESPO-) responsive to diet-induced MFD. Thus, we used RNA-Seq to examine gene expression profiles in mammary cells to elucidate mechanisms underlying MFD in goats and individual variation in the extent of diet-induced MFD. Differentially expression analyses (DEA) and weighted gene co-expression network analysis (WGCNA) of RNA-Seq data were used to study milk somatic cell transcriptome changes in goats consuming a diet supplemented with marine lipids. There were 45 differentially expressed genes (DEGs) between control (no-MFD, before diet-induced MFD) and MFD, and 18 between RESPO+ and RESPO-. Biological processes and pathways such as "RNA transcription" and "Chromatin modifying enzymes" were downregulated in MFD compared with controls. Regarding susceptibility to diet-induced MFD, we identified the "Triglyceride Biosynthesis" pathway upregulated in RESPO- goats. The WGCNA approach identified 9 significant functional modules related to milk fat production and one module to the fat yield decrease in diet-induced MFD. The onset of MFD in dairy goats is influenced by the downregulation of SREBF1, other transcription factors and chromatin-modifying enzymes. A list of DEGs between RESPO+ and RESPO- goats (e.g., DBI and GPD1), and a co-related gene network linked to the decrease in milk fat (ABCD3, FABP3, and PLIN2) was uncovered. Results suggest that alterations in fatty acid transport may play an important role in determining individual variation. These candidate genes should be further investigated.

Milk fat depression and plasma lipids in dairy cows and goats

This study examines the effects of diets supplemented with various lipids selected to induce divergent milk fat content responses (including a milk fat depression) between dairy cows and goats on plasma lipid composition. The objective was to better understand the mechanisms behind the regulation of milk fat secretion in these two ruminant species. Twelve Holstein cows and 12 Alpine goats were fed a basal diet not supplemented (CTL) or supplemented with corn oil plus wheat starch (COS, 5% DM intake (DMI)), marine algae powder of Schizochytrium sp. (MAP, 1.5% DMI), or hydrogenated palm oil (HPO, 3% DMI), in a replicated 4 × 4 Latin square design, during 28 days. On day 27, blood samples were collected for lipid analysis. Plasma lipid classes were quantified by high-performance thin-layer chromatography, with triacylglycerol (TAG) and free fatty acid (FFA) fractions analysed for FA composition by GLC. Plasma molecular species of TAG and ceramides were determined by HPLC–high-resolution MS and by liquid chromatography–triple quadrupole, respectively. Irrespective of diet, plasma total lipid content was higher in cows than goats (+61%), and TAG concentration was higher in goats than cows (+157%). In cows, conversely to goats, COS increased the trans-10 C18:1 proportion in the free FA (+248%) and the TAG (+195%) fractions. In cows and goats, MAP induced increases in cholesterol esters, cholesterol and phospholipids compared to CTL and changes in the plasma free FA and FA of TAG profiles. In both ruminant species, the concentrations of the lipid fractions were unchanged by HPO compared to CTL. Our results point to species specificities and different diet effects in plasma concentrations and compositions of lipid fractions in cows and goats. These new data highlight how diets, that induce large variations in milk fat secretions, affect the plasma lipid classes available for milk fat synthesis.

Milk fatty acids as covariates in multiple regression analysis is a robust approach to model the decrease in milk fat concentration and yield in small ruminants.

Milk fat depression (MFD) syndrome has been associated with the antilipogenic effects of trans-10 fatty acids (FA), such as t10, c12-CLA (CLA) and t10-18:1 (T10). However, these FA alone cannot completely explain the changes in milk fat in small ruminants. Thus, the aim of this study was to use multiple regression analysis to evaluate other FA that may be related to shifts in milk fat, as well as to improve model accuracy when different milk FA are used as covariates in the models. Previously published data were used in multiple regression analysis for goats (n = 106) and ewes (n = 68). Body weight (BW), vaccenic acid (t11-18:1), both trans-10 FA and the major milk FA were tested as covariates to model four response variables associated with MFD: fat concentration (FC), percentage change in milk fat concentration (CFC; %), fat yield (FY; g/d) and percentage change in milk fat yield (CFY; %). All four multiple regression models were significant for both species. When compared with simple regression models, all multiple regression models improved accuracy when estimating MFD. The improvements in model accuracy (lower RMSE) for FC, CFC, FY and CFY were 60.6%, 43.3%, 35.6% and 44.4% for ewes, and 52.1%, 60.1%, 33.6% and 14.9% for goats respectively. Linolenic acid and t11-18:1 were covariates in all models for goats, and palmitic acid and CLA were covariates in all ewe models. These FA should be investigated regarding their direct effect on gene expression associated with milk fat metabolism in the mammary gland of small ruminants. Multiple regression analysis is the most robust approach to account for the variation of milk fat and yield in goats and ewes.

Size, numberand phospholipid composition of milk fat globules are affected by dietary conjugated linoleic acid.

Milk fat globules (MFGs) surround the triacylglycerol core that composes milk fat. The aim of this study isto induce milk fat depression via dietary conjugated linoleic acid (CLA) supplementation to study MFG size parameters, numberand glycerophospholipid composition. Eighteen Holstein dairy cows (136 ± 28 days in milk, 571 ± 37.9 kg body weight, 27.6 ± 2.1 kg milk/day) were selected and randomly assigned to a control or CLA group for a 14-dayperiod. Cows were fed a basal diet (control, n = 8) or the control plus 400 g/day CLA (C18:2 cis-9, trans-11 38.1% and C18:2 trans-10, cis-12 36.8%) (n = 10) for 7 days after which the CLA group was switched to the basal diet for another 7 days along with the control group. Cow performance, milk composition, MFG size and numbers were measured daily. On the seventh day after the start of the experiment, milk samples were identified and the quantification of glycerophospholipid compounds, and RNA were isolated from milk fat samples for a real-time polymerase chain reaction. Compared with control, at Day 7 from the start of feeding, supplemental CLA did not affect milk production (28.09vs. 28.50 kg/day), dry matter intake (14.9vs. 15.4 kg/day), or milk protein (3.55/100vs. 3.70 g/100 ml) and lactose contents (5.11/100vs. 5.17 g/100 ml). However, although the specific surface area of MFG (2138vs. 1815 m²/kg) was greater, CLA reduced milk fat content (1.95/100vs 3.64 g/100 ml on Day 7) and particle size parameters of MFG. The number of MFG gradually decreased until Day 7 of feeding, and then increased by Day 14 (2.96 × 109 on Day 1, 1.63 × 109 on Day 7 and 2.28 × 109 on Day 14) in the CLA group. Compared with control, glycerophospholipid analysis revealed that concentrations of phosphatidylcholine (PC) (e.g., PC [16:0/18:1] 20322vs. 29793 nmol/L), lysophosphatidylethanolamine (LPE) (e.g., LPE [18:1] 956vs. 4610 nmol/L)and phosphatidylethanolamine (PE) (e.g., PE [16:0/18:1] 7000 vs. 9769 nmol/L) in milk lipids decreased during CLA feeding. In contrast, concentrations of phosphatidylinositol (PI) (e.g., PI [18:0/18:1] 4052vs. 1799 nmol/L) and phosphatidylserine (PS) (e.g., PS [18:1/18:2] 9500 vs. 6843 nmol/L) increased. The messenger RNA abundance of fatty acid synthase, diacylglycerol O-acyltransferase 1, glycerol-3-phosphate acyltransferase 4and phosphate cytidylyltransferase 1, choline, alpha (PCYT1A) were downregulated in the CLA group, confirming published data demonstrating a negative effect of CLA on lipogenesis in the mammary gland. Overall, these results provided evidence for the important role of lipogenic gene expression in the regulation of MFG size, number and glycerophospholipid composition.

Milk fat globule membrane proteins are involved in controlling the size of milk fat globules during conjugated linoleic acid-induced milk fat depression.

Milk fat globule membrane (MFGM) proteins surround the triacylglycerol core comprising milk fat globules (MFG). We previously detected a decrease in the size of fat globules during conjugated linoleic acid (CLA)-induced milk fat depression (MFD), and other studies have reported that some MFGM proteins play a central role in regulating mammary cellular lipid droplet size. However, little is known about the relationship between MFD, MFG size, and MFGM proteins in bovine milk. The aim of this study was to investigate the profile of MFGM proteins during MFD induced by CLA. Sixteen mid-lactating Holstein cows (145 ± 24 d in milk) with similar body condition and parity were divided into control and CLA groups over a 10-d period. Cows were fed a basal diet (control, n = 8) or control plus 15 g/kg of dry matter (DM) CLA (n = 8) to induce MFD. Cow performance, milk composition, and MFG size were measured daily. On d 10, MFGM proteins were extracted and identified by quantitative proteomic analysis, and western blotting was used to verify a subset of the identified MFGM proteins. Compared with controls, supplemental CLA did not affect milk production, DM intake, or milk protein and lactose contents. However, CLA reduced milk fat content (3.73 g/100 mL vs. 2.47 g/100 mL) and the size parameters volume-related diameter D[4,3] (3.72 μm vs. 3.35 μm) and surface area-related diameter D[3,2] (3.13 μm vs. 2.80 μm), but increased specific surface area of MFG (1,905 m2/kg vs. 2,188 m2/kg). In total, 177 differentially expressed proteins were detected in milk from cows with CLA-induced MFD, 60 of which were upregulated and 117 downregulated. Correlation analysis showed that MFG size was negatively correlated with various proteins, including XDH and FABP3, and positively correlated with MFG-E8, RAB19, and APOA1. The results provide evidence for an important role of MFGM proteins in regulating MFG diameter, and they facilitate a mechanistic understanding of diet-induced MFD.

Interaction of sodium acetate supplementation and dietary fiber level on feeding behavior, digestibility, milk synthesis, and plasma metabolites.

Acetate supplementation has been shown to increase milk fat yield in diets with low risk of biohydrogenation-induced milk fat depression. The interaction of acetate supplementation with specific dietary factors that modify rumen fermentation and short-chain fatty acid (FA) synthesis has not been investigated. The objective of this experiment was to determine the effect of acetate supplemented as sodium acetate at 2 dietary fiber levels. Our hypothesis was that acetate would increase milk fat production more in animals fed the low-fiber diet. Twelve lactating multiparous Holstein cows were arranged in a 4 × 4 Latin square design balanced for carryover effects with a 2 × 2 factorial arrangement of dietary fiber level and acetate supplementation with 21-d experimental periods. The high-fiber diet had 32% neutral detergent fiber and 21.8% starch, and the low-fiber diet had 29.5% neutral detergent fiber and 28.7% starch created by substitution of forages predominantly for ground corn grain. Acetate was supplemented in the diet at an average 2.8% of dry matter (DM) to provide approximately 10 mol/d of acetate as anhydrous sodium acetate. Acetate supplementation increased DM intake by 6%, with no effect on meal frequency or size. Furthermore, acetate supplementation slightly increased total-tract apparent DM digestibility and tended to increase organic matter digestibility. Acetate supplementation increased milk fat concentration and yield by 8.6 and 10.5%, respectively, but there was no interaction with dietary fiber. The increase in milk fat synthesis was associated with 46 and 85 g/d increases in the yield of de novo (<16C) and mixed source (16C) FA, respectively, with no changes in yield of preformed FA (>16C). There was a 9% increase in the concentration of milk mixed-source FA and a 7% decrease in milk preformed FA with acetate supplementation, regardless of dietary fiber level. Acetate supplementation also increased the concentrations of plasma acetate and β-hydroxybutyrate, major metabolic substrates for mammary lipogenesis. Overall, acetate supplementation increased milk fat yield regardless of dietary fiber level through an increase mostly caused by an increase in longer-chain de novo FA, suggesting stimulation of mammary lipogenesis. The heightened mammary de novo lipogenesis was supported by an increase in the concentration of metabolic substrates in plasma.

Quantitative relationships between ingested and intestinal flows of linoleic and alpha-linolenic acids, body weight and milk performance in mid-lactation dairy cows

Linoleic acid (LA) and alpha-linolenic acid (ALA) are essential fatty acids found in variable quantities in ruminant feedstuffs. Revision of French feed unit systems in 2018 has proposed the reassessment of energy requirements through a between-experiment approach expressing metabolisable energy supply as a function of the energy expenditures for maintenance and production, with these expenditures that reflect homeorhetic regulations. Based on the same approach, LA and ALA intake can be related to animal characteristics (i.e., BW) reflecting maintenance expenditures and secretion characteristics (i.e., milk yield, milk fat content and contents of LA and ALA in milk fat). Therefore, the objective of this work was to analyse the between-experiment relationships between ingested, duodenal, or absorbed flows of LA and ALA, BW and milk LA and ALA secretion by meta-analysis in mid-lactation dairy cows. These relationships were analysed using LA and ALA subsets of 96 and 99 experiments, respectively. Between-experiment regressions of daily flows of ingested, duodenal or absorbed LA and ALA on BW and milk LA and ALA flows were studied, with statistical unit defined as the mean of within-experiment treatments. For LA, the BW-associated coefficient was 0.019 (±0.0034) g absorbed LA/d per kg BW and milk LA secretion-associated coefficient was 0.70 (±0.081) g absorbed LA/g of LA secreted into milk. For ALA, the BW-associated coefficient was 0.0058 (±0.00093) g absorbed ALA/d per kg BW and milk ALA secretion-associated coefficient was 0.57 (±0.097) g absorbed ALA/g of ALA secreted into milk. When coding the diets as either control or milk fat depression diets, the BW-associated coefficient for LA was 0.017 (±0.0032) g absorbed LA/d per kg BW for both diets. For milk fat depression diets, milk LA secretion-associated coefficient was 1.02 (±0.119) g absorbed LA/g of LA secreted into milk, whereas it was 0.70 (±0.075) g absorbed LA/g of LA secreted into milk for control diets. Significant BW and milk performance coefficients were obtained in all LA and ALA equations, allowing the calculation of ingested and intestinal flows of LA and ALA based on measured BW, milk fat yield and milk fat content of LA and ALA. The relationships between ingested and intestinal flows of LA and ALA, BW and milk performance obtained in the present work could be integrated into renewed feed unit systems for energy and protein in dairy cows.

Effects of corn distillers grains with yeast bodies and manipulation of dietary cation and anion difference on production, nutrient digestibility, and gas emissions from manure in lactating cows.

In a randomized complete block design, 40 lactating Holstein cows (average 98 d in milk and 41 kg/d of milk yield) were randomly assigned to 1 of 4 diets: (1) containing soybean meal as the major protein supplement (CON diet); (2) CON diet with high-protein dried corn distillers grains at 20% on a dry matter (DM) basis by replacing mainly soybean meal (DG diet); (3) DG diet except that high-protein dried corn distillers grains with yeast bodies (extracted after corn ethanol production) was used (DGY diet); or (4) DG diet supplemented with sodium bicarbonate and potassium carbonate to elevate the dietary cation and anion difference (DCAD; DG-DCAD diet). The DCAD of CON, DG, DGY, and DG-DCAD were 185, 62, 67, and 187 mEq/kg of DM, respectively. The experiment began with a 10-d covariate period and then cows were fed the experimental diets for 5 wk (2-wk diet adaptation and 3-wk data collection periods). Dry matter intake and milk yield were measured daily, and spot urine and fecal samples were collected in the last week of the experiment to measure nutrient digestibility; N, S, and P utilization and excretion; and in vitro NH3 and H2S emissions from manure. All data were analyzed using the MIXED procedure of SAS (random effect: block; fixed effects: diets, repeated week, and interactions). During data collection, DM intake was not different among treatment groups, but milk yield tended to be lower (42.4 vs. 39.9 kg/d) for DG, DGY, and DG-DCAD versus CON, which could have been caused by decreases in organic matter and neutral detergent fiber digestibility. Milk protein yield tended to be lower (1.33 vs. 1.24 kg/d) for DG, DGY, and DG-DCAD versus CON. Milk fat yield was lower (1.26 vs. 1.55 kg/d) for DG and DGY versus CON, but that for DG-DCAD (1.43 kg/d) did not differ from CON. Similarly, energy-corrected milk was lower (38.0 vs. 43.3 kg/d) for cows on DG and DGY versus those on CON, but it did not differ between DG-DCAD (40.7 kg/d) and CON. Urinary and fecal N excretion were greater for DG, DGY, and DG-DCAD compared with CON due to greater dietary crude protein content and N intake. However, NH3 emissions did not differ across treatments. Intakes of dietary P and S were greater for DG, DGY, and DG-DCAD, resulting in greater excretion of those in manure and greater H2S emissions from manure compared with CON. These data suggest that the negative effects of feeding distillers grains on production of lactating cows can be partly explained by a decrease in nutrient digestibility (milk yield) and excessive anion load (milk fat). The milk fat response to DG-DCAD suggests that milk fat depression observed with a diet with high content of distillers grains can be partially alleviated by supplementation of cations. In the current study, we observed no beneficial effects of DG containing yeast bodies.

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.

In vivo kinetics of oleic, linoleic, and α-linolenic acid biohydrogenation in the rumen of dairy cows.

Ruminal biohydrogenation (BH) of unsaturated fatty acids (FA) reduces absorption of essential FA and can result in formation of bioactive FA that cause milk fat depression. Rates of biohydrogenation of unsaturated FA are commonly observed using in vitro systems and are not well described in vivo. Seven ruminally cannulated cows were enrolled in a 3 × 3 Latin square design study to quantify biohydrogenation of 18:1n-9, 18:2n-6, and 18:3n-3 using a recently developed in vivo BH assay. All cows were fed a common high corn silage basal diet. Biohydrogenation was quantified using a perturbation model that consisted of a bolus dose of 200 g of an oil enriched in each unsaturated FA (oleic acid, OA = 87% 18:1n-9 sunflower oil; linoleic acid, LA = 70% 18:2n-6 safflower oil; and α-linolenic acid, ALA = 54% 18:3n-3 flaxseed oil) and 12 g of 17:0 as a marker of rumen outflow. Rumen contents were sampled before and after the bolus and enrichment of the bolused FA modeled. Using first-order kinetics to model FA disappearance, the fractional rates of disappearance of 18:1n-9 was 0.597 per hour, 18:2n-6 was 0.618 per hour, and 18:3n-3 was 0.834 per hour, similar to rates previously reported with this approach. Rumen turnover of 17:0 was 0.123 per hour, 0.065 per hour, and 0.106 per hour during the OA, LA, and ALA treatments, respectively. The extents of BH were calculated to be 82.8, 90.4, and 88.6% for 18:1n-9, 18:2n-6, and 18:3n-3, respectively. Finally, compartmental modeling was used to quantify the amount of each unsaturated FA metabolized through trans-10 and trans-11 BH pathways. The recently developed in vivo BH assay was able to predict rates of BH and provide insight into rumen metabolism of individual FA and may be useful to future investigations.