Various measurements of microbial productivity in streambed pebble biofilms were analyzed almost monthly for 1 year to quantify the importance of primary production as an autochthonous source of organic matter utilized to support heterotrophic bacterial production in the dynamic food web within this natural microbial habitat. Bacterial density varied from 0.3x10(8) to 1.4x10(8) cells cm-2, and chlorophyll a concentration ranged from 0.7 to 25.9 microg cm-2, with no coupled oscillation between seasonal changes in these two parameters. In bottle incubation experiments, the instantaneous bacterial growth rate of bacteria was significantly correlated with their production rate [measured by frequency of dividing cells (FDC)] as follows: ln mu=0.138FDC-3.003 (n=15, r2=0.445, p<0.001). FDC values in the pebble biofilms increased with fluctuations during the study period, ranging from 3.6% to 9.2%. Bacterial production rates largely fluctuated between 0.15 to 0.92 microg C cm-2 h-1, and its seasonal pattern was similar to that of bacterial density. Net primary production measured between May 2002 to November 2002 attained minimum level (0.5 microg C cm-2 h-1) in June and maximum level (1.9 microg C cm-2 h-1) in August. Percentages of bacterial production to net primary production ranged between 21% and 120%. Because this ratio extends both below and above 100% for these parameters, it is likely that both autochthonous and allochthonous supplies of organic matter are important for production of bacteria in the pebble biofilms that develop in rapidly flowing fresh water streams.

In microbiological studies in streams, pebble samples have until now been taken out of the water following the conventional method. However, this allows the loss of microorganisms as a result of the removal of overlying water. In the present study, to minimize the loss of microorganisms, we have developed a new sampling method, called the submerged method, for collecting microorganisms living on pebbles in streams. The abundance of microorganisms on natural pebbles and artificial clay tiles, both of which had biofilms developing on their surfaces, was measured using both the conventional and submerged methods and the results from the two methods were compared. The cell densities of bacteria (0.10-14.00 ϫ 10 8 cells cm Ϫ2 ), heterotrophic nanoflagellates (0.36-50.30 ϫ 10 4 cells cm Ϫ2 ), and ciliates (0.071-88.27 ϫ 10 2 cells cm Ϫ2 ) measured by the submerged method tended to be higher than those obtained by the conventional method, although there were only a few cases in which a significant difference existed between microbial abundances determined by the two methods. Also supported by microscopic observation, these results suggest the presence of planktonic and/or weakly attached microorganisms on substrate materials in streams. Significant correlations between the concentration of chlorophyll a and the cell densities of heterotrophic microorganisms and significant correlations among heterotrophic microorganisms suggest the presence of active microbial food webs in streams.