Subacute rumen acidosis is thought to be a common condition in early lactating dairy cattle; however, diagnosis is difficult. There are currently only two techniques available for measuring rumen pH under field conditions: rumenocentesis and oral stomach tube. Sixteen rumen-fistulated cows were sampled in four sites of the rumen (cranial-ventral, caudal-ventral, central, and cranial-dorsal) with a rumen cannula. Rumen pH results were compared to those obtained at the same time with rumenocentesis and with an oro-ruminal (Geishauser) probe. Rumen fluid was obtained between 6 and 12 wk of lactation. Samples were analyzed for pH, lactate, bicarbonate, sodium, potassium, and chloride. Rumen pH results were also compared to those obtained from 24-h continuous rumen pH measurement using indwelling rumen pH probes. Oro-ruminal probe samples had the highest pH values and the highest bicarbonate concentrations. Rumenocentesis samples had the lowest pH values and the lowest bicarbonate concentrations. Small differences in electrolyte concentrations were noted among rumen fluid collection techniques in the different rumen sites. The highest correlations of rumen pH were obtained between rumenocentesis and rumen cannulation (cranial-ventral), and between rumen cannulation (cranial-ventral) and the 24-h indwelling pH meter. Compared with samples obtained from the cranial-ventral rumen, rumenocentesis was more sensitive than the oro-ruminal probe in the measurement of low rumen pH; both techniques were moderately specific. The most accurate field technique was rumenocentesis. Improved field techniques are required for better on-farm diagnosis of subacute rumen acidosis.