Abstract: Spray application offers low-cost and efficient application of biologic and reduced concerns regarding diverse water quality and medicator/proportioner function. The objective of the present study was to evaluate the spray application of a Lactobacillus-based probiotic on Salmonella enteritidis (SE) colonization in broiler chickens. Day-of-hatch chicks were challenged with Salmonella enteritidis (SE) by oral gavage alone, challenged with SE and treated by coarse spray application of a commercially-availably Lactic-acid bacterial probiotic (FM-B11™), or challenged with SE and treated with B11 continuously in the Drinking Water (DW). Five days post-challenge, cecal tonsils were collected for presence or absence of SE. In Exp. 1, probiotic treatment by either spray or DW application significantly (p<0.05) reduced SE recovery (55% and 50% respectively; controls 85%) when chicks were held for 8h prior to challenge and placement. Similarly, when probiotic spray treatment or water treatment and challenge occurred simultaneously, with placement 8h after treatment, a marked and significant reduction of SE recovery was noted after 5d (10% and 40% respectively, controls 55%). In Exp. 2, when probiotic spray treatment and challenge occurred simultaneously, with placement 8h after treatment, a significant reduction of SE recovery was again noted in both the spray and DW application (80% controls, 15% spray, 15% DW). Taken together, these results suggest that spray application of B11, when performed in the manner described above, can be effective for protection of chicks against Salmonella infection.