Four experiments were conducted to examine the effect of various factors on in situ digestion of feedstuffs in the rumen of Holstein cows. In Exp. 1, the effect of various days, animals and animal diets on in situ digestion was evaluated. Measured disappearance of dry matter (DM) and nitrogen (N) from soybean meal (SBM) suspended in dacron bags was lowest when the host animal was fed a high grain diet. No effect due to animals, day of incubation or period of experimentation was observed. The effect of bag porosity and substrate particle size on DM and N disappearance from bags was studied in Exp. 2. Rip-stop nylon, a material with small pores, appeared to decrease the influx of digesting agents into bags and limited efflux of digested residues from bags when compared with bags made from dacron polyester material. Disappearance of DM and N from SBM and distillers grains (DG) in rip-stop nylon bags was greater when these feedstuffs were in the pulverized form rather than in the commercially processed form. In Exp. 3, dacron and Acropore bags, with pore sizes of 52 and 5 microns, respectively, were used to examine DM and N disappearance, in the rumen of cows, from total mixed diets containing various dietary N sources. Lower DM and N disappearance from Acropore bags was observed for each of the diets regardless of time spent in the rumen; much of the difference was established within the first hour. The effect of formaldehyde treatment on degradation of amino acids from SBM suspended in dacron bags in the rumen for 16 h was studied in Exp. 4. Increased resistance of SBM amino acids to degradation in the rumen was observed as a result of formaldehyde treatment levels as low as .3% by weight. A possible increase in protection of lysine from degradation relative to other amino acids was also observed.