[In vitro estimation using radioactive phosphorus of the phosphorus requirements of rumen microorganisms]

Microbial requirements for P were assumed to be a function of the amount of microbial protein synthesis (microbial growth) and of the quantity of organic matter (OM) fermented in the rumen. The relationships among P incorporation into microbial matter and protein synthesis, ammonia utilization, volatile fatty acid (VFA) production and organic matter fermented (OMF) were studied in short-term incubations (3 h) using 32P-labelled phosphate. The amount of P incorporated was calculated from extracellular phosphate pool specific activity and the radioactivity incorporated into the microbial sediment during incubation (table 1). The inocula came from sheep fed a protein-free purified diet. In order to vary the intensity of fermentation, carbohydrates with a wide range of degrees of enzymatic susceptibility were used as substrates and the medium was either provided or was deficient in S and trace elements (table 4). Nitrogen was supplied as ammonium salts. Linear regression analyses showed that P incorporation was positively correlated with the criteria of protein synthesis and OM fermentation (figs. 1, 2, 3, 4). However, there was significant phosphorus incorporation when the value for nitrogen incorporation was zero (equation A: (Pi (mg) = 0.162 NH3-N + 0.376; r = 0.9). This was assumed to result either from energetic uncoupling (fermentation without concomitant bacterial growth) or from the lysis of cold microbial cells only. Equation A would reflect total P incorporation and equation A' Pi (mg) = 0.162 NH3-N (mg), net P incorporation. It was assumed that in vitro microbial requirements for P were in the range of 30-70 mg of P/liter of medium for 3-hour incubation, depending on the intensity of fermentation. From a mean value of microbial N yield of 30 g/kg of DOMR (organic matter apparently digested in the rumen), it was calculated that the total and net P requirements in vivo were 6 and 4.9 g/kg of DOMR, respectively, corresponding to 3.9 and 3.2 g/kg of DOM (digestible organic matter). From equation D, relating Pi to OMF, the P requirements were about 4.4 g/kg of DOM. It is suggested that microbial requirements for P varied from 3 to 5 g of P/kg of DOM, depending on the efficiency of microbial synthesis and the extent of carbohydrate fermentation. These results, considered as indicative, should be checked in in vivo experiments.