AND R. E. HUNGATE. Succinic acid turnover and propionate production in the bovine rumen. Appl. Microbiol. 11:132-135. 1963.-High velocity constants for conversion of added succinate to propionate, together with estimations of pool size, showed that extracellular succinate is the major precursor of the propionate formed in the rumen. Some bacteria give off succinate as a final fermentation product which is decarboxylated by others to propionate. Many pure cultures of rumen bacteria form succinic acid as a final fermentation product (Hungate, 1960), but it does not accumulate in the rumen (Sijpesteijn and Elsden, 1952). Added succinate is rapidly converted to propionate in the rumen, by washed rumen bacteria (Sijpesteijn and Elsden, 1952), by Veillonella gazogenes (Johns, 1951a, b), and slowly by species of the genus Propionibacterium (Delwiche, 1948; Johns, 1951c). The object of this investigation was to determine the importance of succinate as a precursor of rumen propionate. This has been attempted by comparing the rate of succinate turnover with the rate of propionate production in rumen contents removed from the animal and incubated in vitro for a brief period. MATERIALS AND METHODS Succinate estimation. The assay procedure was essentially that of Rodgers (1961), using a washed pigeon breast muscle preparation as a source of succinic dehydrogenase and 2,6-dichlorophenol-indophenol as the indicator. The reagents were those described by Rodgers (1961), with the exception that the 2, 6-dichlorophenol-indophenol was purified (Punnet, 1959) and the stock solution (0.15 %; w/v) diluted 15 X with 0.06 M phosphate buffer (pH 7.0) immediately before use. Pigeon breast muscle was prepared, washed (Rodgers, 1961), and weighed out in 5-g amounts onto squares of aluminum foil. These were closed up and stored in a screwcap jar at-15 C for up to 6 months. Before use, the washed muscle (5 g) was homogenized for 1 min in a Waring Blendor in 60 ml of 0.