An eight-week feeding trial was conducted to examine the impact of dietary supplementation with glycerol monolaurate (GML) on juvenile black sea bream. A basal diet was formulated containing 24% fish meal, while five additional diets were prepared, each supplemented with varying levels of GML: GML1 (0.01%), GML2 (0.02%), GML3 (0.04%), GML4 (0.08%), and GML5 (0.16%). Triplicate tanks were randomly allocated to each diet, each containing 20 fish with an initial weight of 1.55 ± 0.05 g. By the trial's end, the GML3 group displayed a notably higher final body weight (FBW), weight gain (WG), specific growth rate (SGR), and protein efficiency ratio (PER) compared to the other groups (p < 0.05), but the FCR was significantly higher in the control group. However, no significant differences were observed in the MFI, PPV, CF, HSI, IPF, VSI, or SR among the groups (p > 0.05). Regarding the proximate compositions of the dorsal muscle and whole body, no substantial differences were observed across the groups (p > 0.05). Additionally, there were no significant variations in digestive enzyme activity (p > 0.05), serum immune, or biochemical parameters in the midgut and hindgut among the treatment groups. But in the serum immune response IgM, C3 and C4 were significantly higher in the GML3 group as compared to the other groups (p < 0.05). However, the GML3 group exhibited significantly greater fore-intestinal villus height, crypt depth, villus height per crypt depth, and the number of goblet cells per villus compared to the other groups (p < 0.05). Overall, GML supplementation, particularly GML3, significantly improved growth indicators like the final body weight and intestinal morphology. While certain parameters remained unaffected, these findings suggest GML's potential as a beneficial dietary supplement in fish diets.