Invasion of gingival tissues by bacteria is thought to be a major factor in development of periodontal lesions. Morphologic studies have revealed bacteria within the pocket epithelium, gingival connective tissues, alveolar bone, and oral epithelium. The current studies are intended to determine whether they are present in healthy and diseased tissues and to identify the microbial source. Five beagle dogs with naturally occurring periodontitis were fed a soft diet. Two quadrants of the dentition of each dog were cleaned regularly and health maintained while others were allowed to develop further disease. After 4 weeks, samples of gingiva were removed both prior to and after the animals were sacrificed by perfusion fixation attempting not to introduce microorganisms. The identical samples were examined by light and scanning electron microscopy for numbers and location of organisms. Their numbers were greater in diseased than in healthy tissues. Furthermore, the quantity was greater in samples taken prior to perfusion fixation than after such treatment, especially in the oral epithelium. The results suggest that normally there are some microorganisms present within the periodontal tissues and that their numbers are greater in diseased tissues. However, the data also indicated that significant numbers may be introduced into the tissues during sampling of unfixed tissues.