I . Four feeding trials were made to investigate relations between diet and protozoal population in the rumen.2. When a ration containing no concentrate was used, the number of entodiniomorphs decreased rapidly. The number of entodinioniorphs increased with the amount of concentrate.Rice straw, which was used as a sole source of roughage, was not always necessary for protozoa to survive in the rumen.3. The type of diet affected the holotrich population to a much smaller extent than the entodiniomorph population.During the past two decades there has been a gradual accumulation of information about the factors that influence the number of protozoa in the rumen (Warner, 196;; Hungatc, 1966;Eadie & Mann, 1970). For example, the physical form of ration and the frequency of feeding have generally been accepted to influence protozoal concentrations in the rumen. However, much more detailed work will be needed.The present work describes four experiments designed to investigate the influence of the type and amount of diet on protozoal concentrations in the rumen. E X P ER I IVI ENT A L Feeding trialsThree Holstein cows with permanent rumen fistulas, and weighing approximately 500 kg each, were used in four experiments. Before each experiment, they were given a basal ration (B) and then other rations (1-6) as shown in Table I . They were fed twice daily at 08.00 and 16.00 hours and allowed free access to water.Expt I. Effects of three rations on protozoal population in the rumen were examined. Three cows were used in a 3 x 3 Latin square design as shown in Table I. The three treatment periods were 5 d each. As ration z contained no roughage and was generally poor, each treatment period was followed by 10 d intervals on ration B to restore the condition and appetite of the animals. On the last days of three treatment periods, samples of rumen fluid were collected at 15.30 hours through the fistula by the use of a vacuum pump and a rubber tube.
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