The effects of step-wise improvement of forage combination in total mixed rations on fatty acid profile in the rumen and milk of Holstein cows

Five lactating Holstein cows in a 5×5 Latin square experiment were fed five high-concentrate total mixed rations (TMRs) to investigate the effects of step-wise improvement of forage combination on ruminal and milk fatty acid profiles. The ratio of concentrate to forage was fixed as 61:39, and the step-wise improvement of forage combination was applied as: TMR1, a ration containing corn stover; TMR2, a ration containing corn stover and ensiled corn stover; TMR3, a ration containing ensiled corn stover and Chinese wild ryegrass hay (Leymus chinensis); TMR4, a ration containing the ryegrass hay and whole corn silage; TMR5, a ration containing the ryegrass hay, whole corn silage and alfalfa hay. The TMRs were offered to the cows twice daily at 0700 and 1900 h. The entire experiment was completed in five periods, and each period lasted for 18 days. Diurnal samples of rumen fluids were collected at 0100, 0700, 1300 and 1900 h (day 16); 0300, 0900, 1500 and 2100 h (day 17); and 0500, 1100, 1700 and 2300 h (day 18). The step-wise improvement of forage combination increased energy and crude protein contents and decreased fibre content. As a result, the step-wise improvement of forage combination increased dry matter intake and milk yield (P