The genus Propionibacterium consists of two principal groups, cutaneous and classical or dairy. Cutaneous species are predominant members of the microbial population of human skin and have also been isolated from the feces of humans and other vertebrate animals. They are often considered opportunistic organisms and have been occasionally associated with infections in humans. Dairy propionibacteria are microorganisms extensively used in the industry for manufacture of Swiss-type cheeses and biological production of propionic acid and vitamin B12. They can be isolated from soil, vegetables, silage, raw milk, and dairy products such as kefir and different cheeses with "eyes."In the last decade, several studies have demonstrated probiotic properties for members of the genus Propionibacterium. The effects claimed are based on the production of bacteriocins, vitamins, stimulation of growth of other colonic bacteria like bifidobacteria, beneficial modification of the composition and metabolic activities of the intestinal microflora, immunomodulation, and antimutagenic activity. It is thought that to produce many of these health benefits, the probiotic microorganisms must be able to survive the transit through the hostile conditions of the gastrointestinal tract (GIT) and remain at high levels in the intestine, avoiding removal by peristaltic contractions of the gut. In this sense, microorganisms with a short generation time or the ability to adhere to the intestinal mucosa will survive for prolonged periods in the body of the host. Therefore, two desirable properties for probiotic microorganisms are (1) resistance to gastric acidity, bile, and pancreatic enzymes; and (2) adhesion ability to mucosal surfaces. Dairy bacteria are traditionally not considered to persist as normal inhabitants of the human intestinal tract. Therefore, survival under GIT conditions and adherence are important properties to be considered, and tests to study them would be useful tools. In the present chapter we describe the methods used in our laboratory to assess survival, metabolic activity, and adhesion of dairy propionibacteria to intestinal epithelial cells after gastrointestinal digestion.