Antagonistic effect of piglet microflora against Clostridium perfringens was studied in germfree mice, to isolate bacterial strains responsible for this colonization resistance. The 1:100 dilution of the feces of a 2 day-old conventional piglet, given per os to germfree mice already harboring C. perfringens, led to the elimination of C. perfringens. From this piglet flora, 8 bacterial strains were selected, belonging to the genera Bacteroides, Clostridium, Eubacterium, Bifidobacterium, Lactobacillus and a strain belonging to the class of Mollicutes. When the 8 strains were given to germfree mice 3 days after C. perfringens inoculation, they led to rapid elimination of C. perfringens from feces. Sixteen other mixtures of 2 to 7 strains were similarly tested, but none was able to fully antagonize C. perfringens. When the 8 strains were given per os to germfree piglets after C. perfringens inoculation, they led to the rapid elimination of C. perfringens from pig feces, and to a quick recovery from diarrhea. This study led to the identification of a simplified fraction of gut microflora, able to exert a barrier effect against C. perfringens comparable to the entire flora of the piglet. This study suggests that gnotoxenic mice can be a suitable model for simplifying the flora responsible for a given effect in another host, animal or human.