The fundamental understanding of the mechanisms regulating milk protein synthesis is limited. This study aimed to elucidate the metabolic mechanisms of milk production affected by forage quality through studying metabolites from four biofluids (rumen fluid, milk, serum, and urine) collected from 16 lactating cows fed alfalfa hay (AH, high-quality, n = 8) and corn stover (CS, low-quality, n = 8) using gas chromatography-time-of-flight/mass spectrometry. The cows fed AH exhibited higher milk yield (P < 0.01), milk protein yield (P = 0.04), and milk efficiency (P < 0.01) than those fed CS. A total of 165, 195, 218, and 156 metabolites were identified in the rumen fluid, milk, serum, and urine, respectively, while 29 metabolites were found in all four biofluids. In addition 55, 8, 28, and 31 metabolites in each biofluid were significantly different (VIP > 1 and P < 0.05) between the AH- and CS-fed animals. These metabolites were involved in glycine, serine, and threonine metabolism; tyrosine metabolism; and phenylalanine metabolism. Further integrated key metabolic pathway analysis showed that the AH-fed cows may have more comprehensive amino acid metabolisms, suggesting that these metabolite-associated pathways may serve as biomarkers for higher milk yield and better milk protein quality.

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The objective of the current study was to investigate the effect of N-carbamoylglutamate (NCG) supplementation on milk production and nitrogen (N) utilization in Chinese Holstein dairy cows. Sixty multiparous cows (78±17.3 d in milk, 635±61.00kg of body weight, and 41.9±7.9kg/d milk yield; mean ± SD) were blocked by parity, days in milk, and milk yield and randomly allocated to 1 of 4 groups, each of which was fed a dietary treatment containing 0 (control), 10, 20, or 30g of NCG/d. Milk yield was recorded weekly. Dry matter intake, milk composition, plasma variables, and urea N contents in plasma, urine, and milk were determined every other week. Blood samples were collected from the coccygeal vein. Rumen microbial protein synthesis was estimated based on the purine derivatives in the urine. Dry matter intake was found to be similar between the treatments. Addition of 20g of NCG/d tended to increase milk yield (40.2 vs. 38.1kg/d) and increased the content (2.83 vs. 2.74%) and yield (1.12 vs. 1.02kg/d) of milk protein compared with the control. The yield and content of milk fat were similar between the treatments, whereas the contents of lactose and total solids increased linearly with an increase in NCG. Dietary supplementation of NCG linearly increased the plasma nitric oxide level and decreased the plasma ammonia N level. Compared with the control, the plasma Arg concentration in cows fed 10, 20, and 30g of NCG/d was increased by 1.1, 10.4, and 16.0%, respectively. The urea N concentrations in the milk, plasma, and urine decreased with the addition of NCG, although the lowest urea N concentrations were observed with the addition of 20g of NCG/d. The conversion of dietary crude protein to milk protein exhibited quadratic trends of improvement by NCG supplementation, with a peak at 20g of NCG/d. The rumen microbial protein synthesis was not altered by NCG supplementation, but the metabolizable protein tended to show a quadratic increase in cows fed 20g of NCG/d. In conclusion, supplementation of 20g of NVG/d may alter the plasma metabolites, optimize the AA profile, increase the metabolizable protein utilization, and thereby improve the lactation performance and N utilization of high-yielding dairy cows.

The objective of this experiment was to determine the degradation of L-arginine (ARG) and N-carbamoyl glutamate (NCG) and to examine their effect on rumen fermentation. Rumen fluids were collected from 3 rumen-fistulated cows and then incubated with ARG or NCG at 1 mmol/L in a glass syringe system at 39°C for 24 h. The control treatment was given neither ARG nor NCG. Gas production (GP) was recorded, and pH at 2, 4, 6, 12, and 24 h was also determined. At 12 and 24 h, the measurements were also made for ammonia-nitrogen (N), volatile fatty acids (VFA) and microbial crude protein (MCP) yield on purine quantification basis. At 24 h, the proportion of ARG and NCG degradation in rumen fluid was 100.0 and 17.8%, respectively. Gas production and the acetate to propionate ratio increased in groups treated with ARG and NCG, compared with the control (P<0.01). Ammonia nitrogen concentration was higher (P<0.01) in the ARG group than in the NCG and control groups. Microbial crude protein concentration diminished in ARG and NCG groups, in comparison with the control (P<0.01). In conclusion, the effects of ARG and NCG on rumen fermentation were numerically relatively similar. Rapid degradation of ARG in rumen is a nutritionally wasteful process. Thus, ARG should be spared from rumen degradation, while NCG could be fed to ruminant without need for coating.