An experiment was conducted to examine effects of supplemental lysophospholipids (LPL) in dairy cows. Eight ruminally cannulated lactating Holstein cows were used in a replicated 4 × 4 Latin square design. Dietary treatments were (1) a dairy ration [CON; 55% forage and 45% concentrate on a dry matter (DM) basis], (2) a positive control diet supplemented with monensin (MON; 16 mg/kg in dietary DM; Elanco Animal Health, Greenfield, IN], (3) a control diet supplemented with low LPL (0.05% of dietary DM; Lipidol Ultra, Easy Bio Inc., Seoul, South Korea), and (4) a control diet supplemented with high LPL (0.075% of dietary DM). Experimental periods were 21 d with 14-d diet adaptation and 7-d sample collection. Daily intake and milk yield were measured and rumen contents were collected for fermentation characteristics and bacterial population. Spot urine and fecal samples (8 samples/ cow per period) were collected to determine nutrient digestibility and dietary N utilization. All data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC; group and cow within group were random effects and treatments, time, and their interaction were fixed effects). Preplanned contrasts were made to determine effect of MON versus CON, effect of LPL versus MON, and linear effect of increasing LPL. In the current study, responses to MON generally agreed with effects of monensin observed in the literature (increased milk yield and feed efficiency but decreased milk fat content). Supplementation of LPL to the diet did not alter DM intake but linearly increased milk yield, resulting in increases in feed efficiency (milk yield/DM intake) and milk protein and fat yields. However, total-tract digestibility of DM and organic matter tended to be lower (60.9 vs. 62.2% and 61.8 vs. 63.1%, respectively) for LPL compared with CON. Linear increases in milk N secretion and decreases in urinary N excretion were observed with increasing LPL in the diet. A slight decrease in acetate proportion in the rumen for LPL was found. Relative to MON, very few bacteria in the rumen were affected with increasing LPL. In conclusion, LPL is a potential feed additive that can increase milk yield and components and dietary N utilization. However, more studies with large numbers of animals are needed to confirm the effect of LPL on production. Similar positive effects on production were observed between LPL and MON, but individual mechanisms were likely different according to ruminal fermentation characteristics. Further studies are needed to explore the mode of action of LPL in dairy cows.