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Four lactating Holstein cows were used in a 4횞4 Latin-square design to determine the effects of various levels of whole cottonseed (WCS) in diets on parameters including milk (yield and fat content), rumen fluid (pH, ammonia and TVFA) and blood (棺-carotene, vit. A, vit. E, urea, NH3, Ca, P and Mg levels). Cows consumed 0, 1, 2 or 3 kg WCS per day. No significant differences were observed among the groups on analysed parameters except plasma vitamin E concentration. In addition, when the amount of cottonseed was increased, milk yield and milk fat content also tended to increase but this increase was not statistically significant. In conclusion, feeding of WCS up to 3 kg per day with ad libitum maize silage did not cause negative effects on milk yield, milk fat and blood vitamin levels in the short term in dairy cows.