The factors affecting the DAPI (4', 6-diamidino-2-phenylidole) fluorescence direct count in the tidal river sediment were examined. Sediment samples were collected from the Guangzhou section of the Pearl River. Besides sediment texture and organic matter, an improved staining procedure and the involved parameters were analyzed. Results showed that the procedure with the sediment with 2000 fold dilution and ultrasonic water bath for 10 min, and with a final DAPI concentration of 10 microg x mL(-1) and staining time for more than 30 min produced the optimum results of DAPI direct count in the sediment. The total bacterial number was correlated to the proportion of the non-nucleoid-containing cells to the total bacterial number (r = 0.587, p = 0.004). The organic matter content also correlated to the ration. The clay content had a strong correlation with the organic matter, through which the clay content also affected the ratio. A multiple regression analysis between the ration versus the organic matter, the total bacterial number, and the clay content showed that the regression equation fit the measure values satisfactorily (r = 0.694). These results indicated that the above factors needed to be considered in the applications of the DAPI fluorescence direct counting method to the tidal river sediment.