The purpose of this study was to elucidate 19 enzymatic activities in whole mixed saliva and their changes in men without oral hygiene.Ten healthy, unmedicated male students with excellent oral hygiene and clinically normal gingiva, between the ages of 22 and 26 years, were the experimental subjects. At the start of the experimental period, whole mixed saliva and parotid saliva were collected and the clinical parameters were recorded for each of them. Then, the experimental no-cleansing period of 7 days was started, and whole mixed saliva was collected and the clinical parameters were recorded every day. Enzymatic activities were determined using API ZYM systems (MONTALIEU VERCIEU, France).Alkaline phosphatase, butyrate esterase, caprylate esterase-lipase, myristate lipase, leucine aminopeptidase, valine aminopeptidase, and trypsin activities in whole mixed saliva supernatant were significantly higher at the end of the no-cleansing period than at the start of experiment. Myristate lipase, valine aminopeptidase, and trypsin were detected in whole mixed saliva sediment, but not in parotid saliva. This suggests that these enzyme activities in whole mixed saliva are useful in estimating the oral hygiene status.