Abstract Characteristics of the lactobacilli colonizing the various regions of the porcine gastrointestinal tract were investigated. The lumenal contents from the gastric, jejunal, cecal and colonic regions, and biopsies from the gastric non-secreting and secreting regions were homogenized and serial dilutions were spread on Rogosa agar and colonies (50 per region and animal) were randomly sampled. The lactobacillus isolates were grouped according to their protein profiles of lysozyme-treated whole cells. Several different groups of lactobacillus could be detected. Specific groups were associated and often unique for the stomach, jejunal, cecal and colonic regions of the gastrointestinal tract. A few groups were uniformly distributed through the tract. Although published studies of lactobacilli colonizing stomach mucosa have been restricted mainly to the non-secreting region, significant levels were also detected on the secreting stomach tissue. Population densities ranged from 2 × 10 4 to 2 × 10 6 CFU per cm 2 on both the secretory and non-secretory regions. Of the isolates which colonized the gastric secreting epithelium, 26% formed smooth colonies and 74% formed rough colonies. In contrast, the non-secreting epithelium was mainly colonized by lactobacilli forming smooth colonies (67%). From this study, one can suggest that different lactobacilli are associated with specific regions of the gastrointestinal tract, and that some specific groups appear to be restricted to defined regions.