The objective of this study was to clarify the effect of different milking frequencies under an automatic milking system (AMS) on milk yield, plasma metabolite profiles and mammary arterial-venous (A-V) differences of milk precursors by mammary tissues in early lactation cows. Twelve Holstein cows were divided into two and four times milking frequency treatments by AMS after calving to 50 days postpartum. Cows were given a partial mixed ration ad libitum and a concentrate diet at every milking. Dry matter intake increased similarly in both treatments with advancing postpartum days. Milk yield was greater (P < 0.001) by 25% with four times milking, but milk composition was not affected by milking frequency. Body weight change was also not affected by milking frequency. Arterial concentrations of glucose and glutamate were lower (P < 0.05) for four times milking frequency. However, arterial concentration of nonesterified fatty acids did not differ between treatments. Although mammary A-V differences of plasma concentration for most milk precursors did not differ between treatments, estimated plasma flow was higher (P < 0.05) for four times milking frequency. These results indicate that higher milking frequency may increase mammary uptake of milk precursors, whereas may not affect the extent of fat mobilization of early lactating cows from day 20 postpartum onward.

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