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There is an increasing trend in milk urea concentration has been used as an indicator for monitoring protein nutrition in ruminants because of linear relationship between milk urea concentration and dietary crude protein level. Milk urea analysis is easy to monitor and can be performed on buffaloes and has some advantageous under field conditions for estimating protein nutrition status of buffaloes. Thus, the objective of this study was to investigate the effect of nutrition status on milk urea concentration and milk protein content, and to monitor protein nutrition by determining milk urea nitrogen concentration and milk protein in Anatolian buffaloes. Ten lactating (in the 3rd lactation and 15±3 days in milk) Anatolian buffaloes having approximately accordant live body weights were used in this study. Milk samples were collected biweekly to determine the milk urea concentration and milk nutrients. Individual samples of each feedstuffs consumed by lactating buffaloes were collected biweekly during the experiment. There was no change in milk fat, total solid, lactose and ash during the study. There was a positive correlation (P<0.001) not only between diet crude protein and milk urea concentration, but also between diet crude protein and milk protein values. There was a negative correlation (P<0.001) between energy value of diet and milk urea concentration, and also between energy value of diet and milk protein value. In conclusion, milk urea and protein analyses may provide an opportunity to get knowledge about dietary protein intake.