Five dairy cows were arranged in a 5 x 5 Latin square design to compare the effects of two amounts of either duodenal glucose or ruminal propionic acid (C3) on milk yield and composition. Treatments consisted of a grass silage-based diet supplemented with glucogenic nutrients either infused in the rumen as a mixture of volatile fatty acids (control) or pure C3 (1.72 and 3.45 Mcal/d) or in the duodenum as glucose (1.72 and 3.45 Mcal/d). Treatments were isoenergetic and isonitrogenous and contained 100 and 115% of energy and protein requirements according to INRA (1989), respectively. Only C3 treatments significantly modified ruminal volatile fatty acid composition and linearly increased C3 percentage (up to 25.5%). Both treatments substantially decreased milk fat yield and content, and linearly increased milk and protein yields. Although no significant differences between glucose and C3 were highlighted for milk yield and composition, it seems that mechanisms involved in milk fat decrease are different. Indeed, whereas C3 treatments decreased fatty acid production in an homogeneous way, short- and long-chain fatty acids decreased and medium-chain fatty acid production increased with glucose treatments. A bibliographical study confirmed that increasing glucogenic precursors (GP) supply curvilinearly increase milk yield, linearly increase milk protein content (+ 0.04% per Mcal of GP) and curvilinearly decrease milk fat content (- 0.14% per Mcal of GP). Thus, it appears important to account for the nature of energy supplied by the ration in formulation.
The effect of intestinal glucose supply on whole body rate of glucose appearance (WBGRa) and mammary utilization of glucose was studied in four lactating dairy cows. Glucose (0, 443, 963 and 2398 g/d) was continuously infused in the duodenum over 14-d periods using a Latin square design. A grass silage-based diet was formulated so that treatments were isoenergetic and isonitrogenous and contained 100 and 110% of energy and protein requirements according to INRA (1989). The WBGRa was measured by the [6,6-(2)H2]glucose dilution technique, and mammary glucose balance by arteriovenous differences and blood flow measurements. Duodenal glucose infusion increased arterial glucose concentrations linearly, whereas arterial concentrations of insulin, growth hormone, and glucagon were not changed. The WBGRa increased linearly with increasing glucose loads. The increase represented 42% of the intestinal glucose supplement. Mammary blood flow dramatically increased (up to 45%) and was associated with a significant increase of arterial insulin-like growth factor-1 concentrations. Mammary gland rate of glucose disappearance ([6,6-(2)H2]glucose measurement) increased linearly, whereas net mammary balance of glucose, lactose, and milk yields increased quadratically. Net mammary balance of glucose accounted for 60% of WBGRa, except for the greatest dose (47.6%). The decrease in milk yield with 2398 g/d of glucose may be explained by an imbalance in intracellular intermediate concentrations. The milk ratio of glucose-1-phosphate to glucose-6-phosphate decreased significantly at the greatest infusion of glucose. In conclusion, exogenous glucose supply to a grass silage-based diet increased WBGRa, mammary utilization of glucose and milk synthesis.
Five multiparous Holstein cows were used in a study with a 5 x 5 Latin square design to measure the effects of postruminal infusion of Met on lactational performance and plasma metabolites. The treatments were duodenal infusions of 1) 10 g/d of Lys (control), 2) 10 g/d of Lys plus 6 g/d of Met, 3) 10 g/d of Lys pus 12 g/d of Met, 4) 10 g/d of Lys plus 18 g/d of Met, and 5) 10 g/d of Lys plus 24 g/d of Met. The cows were fed a diet of 61% maize silage, 31% concentrate, and 5% dehydrated alfalfa. The DMI were similar among treatments. Milk yield, 4% FCM, and milk fat yield and content were not affected by infusions. In contrast, milk protein yield and content were increased linearly as Met infusion increased, which was true also for plasma Met and Cys concentrations. Using measurements of AA flow to the duodenum and assumed intestinal digestibilities of 0.8 for digesta and 1.0 for infused AA, estimated concentrations of Lys and Met in total AA absorbed in the small intestine were 7.3% for Lys and 1.52, 1.73, 1.94, 2.15, and 2.36% for Met for diets 1 through 5, respectively. The substantial linear increases in milk protein yield and content indicated that postruminal Met supply was not adequate over the entire range of Met infusions. In conclusion, the extent of Met limitation in this study could be defined only as that exceeding 2.4% of total AA absorbed in the small intestine.
Although in dairy cows the mammary gland (MG) is the major net user of essential AA (EAA) supply, milk protein synthesis from absorbed EAA is not a straightforward process. Early studies identified 2 groups of EAA based on different pattern of mammary utilization: group 1 [Met, Phe (+Tyr), Trp], where MG uptake was similar to secretion in milk protein, and group 2 (Arg, Ile, Leu, Lys, Thr, and Val), where uptake exceeded milk protein output. This review examines the validity of this classification under variable protein supply through a meta-analysis, with the outcomes then explained with studies in which the fates of individual EAA were monitored using isotope approaches. For the meta-analysis, the Fick principle, based on stoichiometric transfer of Phe+Tyr uptake to milk protein, was used to estimate mammary plasma flow across all studies. This approach was judged acceptable because doubling Phe supply did not result in mammary oxidation of Phe+Tyr and either limited or no contribution of peptides to Phe and Tyr mammary supply could be detected. The AA content of proteins synthesized by the MG was estimated from milk protein composition, and the uptake-to-output ratio (U:O) for individual AA was re-calculated based on these assumptions. Analysis of individual samples by isotopic dilution resulted in reduced variance compared with analysis on pooled samples performed with an AA analyzer. Globally, the U:O of His and Met is maintained close to unity under variable protein supply. The group 2 AA could be subdivided. First, the U:O for group "2v" AA (Ile, Leu, Val, and Lys) is greater than 1 and varied with protein supply. Accordingly, the increased U:O of Leu, induced by duodenal casein infusion, led to extra-mammary Leu oxidation. Decreasing Lys supply decreased Lys U:O and the associated transfer of N to non-EAA, mainly to Glx, Asx, Ser, and Ala. Second, the U:O of group "2nv" AA, Arg and Thr, does not vary with protein supply. The Arg U:O averages 2.5, whereas the Thr U:O, albeit averaging 1.2, does not differ from unity. Excess of both these AA is probably directed toward the synthesis of non-EAA rather than energy supply. Overall, the ability of the MG to use excess EAA-N supply offers alternative sources of N and C for energy provision, lactose synthesis and non-EAA synthesis. The latter function spares dietary non-EAA for other necessary processes, such as gluconeogenesis and energy supply, in other tissues to support lactation.
The effects of casein (CN) and propionate (C3) on mammary AA metabolism were determined in 3 multiparous Holstein cows fitted with both duodenal and ruminal cannulas and used in a replicated Youden square with six 14-d periods. Casein (743 g/d in the duodenum) and C3 (1,041 g/d in the rumen) infusions were tested in a factorial arrangement. For each period, L-[1-(13)C]Leu (d 11) and NaH[13C]O3 (d 13) were infused into a jugular vein, and blood samples were taken from the carotid artery and the mammary vein to determine Leu kinetics and net uptake of AA. Both CN and C3 treatments separately increased milk protein concentration and yield. With CN there was a general response in mammary protein metabolism, involving increases in Leu net uptake (30%), the uptake:output ratio (8%), protein synthesis (11%), secretion in milk protein (21%), and oxidation (259%). In contrast, C3 treatments tended to increase only Leu in milk protein (7%) and, when in combination with CN, to reduce Leu used for protein synthesis (5%). Across all treatments, most Leu uptake by the mammary gland was accounted for as Leu in milk or oxidized, and the Leu balance was therefore achieved without involvement of either net peptide use or production. Mammary uptake of group 1 AA increased to match milk output with all infusions. In contrast, mammary uptake of group 2 AA exceeded output to a greater extent with CN than with C3 infusions, whereas the increment in uptake of group 3 AA increased with C3 treatments. Overall, these data suggest that different mechanisms operate to improve milk protein production when either protein or energy is supplied.
Providing a well-balanced supply of essential AA (EAA) can serve as an opportunity to reduce the protein intake for dairy cows by increasing the efficiency of metabolizable protein (or PDIE, its equivalent in the INRA feeding system) utilization for milk protein yield. Our objectives were to compare the effect of supplying an "ideal" EAA profile (EAA+) with an imbalanced AA profile (control) at 2 levels of PDIE/NE(L) (net energy for lactation) supplies to study the interaction between PDIE and AA profiles, and to compare this ideal profile with a simple mixture of the 4 most deficient EAA (4 EAA) in the diets of dairy cows. Six lactating multiparous Holstein cows received 6 treatments with 2 different levels of PDIE supplied by diets and AA infusions in the duodenum according to a changeover design with 3-wk periods. Within each PDIE supply level, the cows received 3 different AA infusions in the duodenum according to a 3×3 Latin square design with 1-wk subperiods, which corresponded to the following treatment groups: control (Glu), 4EAA (Lys, Met, His, Leu), and EAA+ (4 EAA plus Ile, Val, Phe, Trp, and Tyr). In the low and high PDIE treatments, diets and infusions provided 54.7 and 64.0 g/Mcal of PDIE/NE(L), respectively, which corresponded to crude protein levels of 13.6 and 15.2%, respectively. High-PDIE supplies increased the milk protein yield by 163 g/d, the milk protein content by 1.4 g/kg, the milk yield by 4.1 kg/d, and the lactose yield by 178 g/d and decreased the PDIE efficiency of utilization by 12.4%, whereas the N efficiency of utilization remained unaffected. Supplying the 2 EAA profiles (4EAA and EAA+) increased the milk protein yield by 67 g/d, the milk protein content by 1.3g/kg, and the milk yield by 0.9 kg/d, whereas the milk fat and milk lactose contents were decreased by 2.4 and 1.6g/kg, respectively. The responses regarding milk yield and its composition were similar whether the cows received the 4 EAA or the EAA+ treatment. The responses were similar for the milk yield and composition whether the EAA were supplied by low- or high-PDIE supplies. In conclusion, the efficiency of PDIE utilization was improved by 6.6% and the N efficiency was improved by 7.0% by correcting the EAA profiles, independent of the level of PDIE supplied. In addition, the increased efficiency observed, associated with provision of the 4 EAA, was similar to the provision of all EAA (EAA+) in this experiment.
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