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An experiment was conducted to test the hypothesis that the supplementation of crude protein (CP) results in rumen acidosis and increased histamine concentrations in dairy cows. Six ruminally fistulated, non-pregnant dry cows were fed three experimental rations in a replicated 3 × 3 Latin square design. The CP contents in the low-CP, the high rumen undegradable protein (high-RUP) and the high rumen degradable protein (high-RDP) rations were 112, 259 and 266 g/kg dry matter (DM) respectively. The cows were fed 7.7 kg DM of the concentrates and 2.7 kg DM of rice straw. High levels of RDP in the ration significantly increased the ammonia, total volatile fatty acid (VFA) and histamine concentrations in the rumen fluid. However, supplemental CP, whether degradable or undegradable, did not significantly affect the pH of rumen fluid. Plasma urea nitrogen concentration was higher in both high-RDP and high-RUP in relation to cows fed the low-CP ration. The rise in ruminal histamine concentrations was physiologically non-relevant, most likely because rumen pH was not affected by supplemental CP at the installed level of DM intake. Therefore, it can be concluded that the issue of supplemental CP, rumen pH and ruminal histamine concentrations has not yet been settled. Further research is warranted to understand these relationships., (© 2011 Blackwell Verlag GmbH.)