The use of ribonucleic acids as markers for the measuring of microbial protein yield in the rumen. 2. The effect of sample treatment, the time of sampling and the composition of the ration on the RNA:N ratio in rumen microbes]

In two experiments the influence of the treatment of samples, the sampling time and the composition of the rations on the RNA: N ratio in the rumen microbes was checked. Experiment I proved that freezing (-21 degrees C), thawing and freeze-drying of isolated bacteria and protozoa from the rumen fluid and from the duodenal content did not result in a change of the RNA content and the RNA-N: total N relation. If, however, the rumen fluid is stored deep-frozen before the isolation of the bacteria the N content in the DM of the bacteria decreases by 17% and that of RNA by 30%. This results in a change of the RNA: N relation of 16%. In conclusion, the bacteria are to be isolated immediately after rumen fluid sampling. Isolated bacteria can be stored deep-frozen before RNA determination and then freeze-dried. Experiment II showed that the RNA content of the rumen protozoa varies according to the period after feeding. The RNA: N relation was 0.50, 0.92, 0.70 and 0.58 on average 0, 3, 6 and 8 h after feeding, in which the 3rd hour after feeding can obviously be considered the time of increased microbial activity. The conclusion from this variation is that more than one isolation of microbes must be carried out in the course of the day in order to achieve representative samples. These statements apply to easily and not easily fermentable protein as N source in the feed. It could also be proved that no essential variation is to be expected in the RNA: N relation in the microbes isolated from the rumen fluid in the range of 8-21% crude protein in the DM of the ration (roughage: concentrate = 55: 45). On average the rumen microbes contained 1.7 g RNA-N/16 g N, essential differences between bacteria and protozoa could not be ascertained. From the slight variation of the RNA-N: N relation in the isolated bacteria from various cows one can conclude that there is no need to isolate the microbes of each individual animal.