Heterotrophic bacterioplankton abundance and production were investigated with depth (down to bathypelagic layers) and with longitude (from 4.9 degrees E to 32.7 degrees E) along a cruise track across the Mediterranean Sea in early summer 2008. Abundances and flow cytometric characteristics (green fluorescence and side scatter signals) of high nucleic acid (HNA) and low nucleic acid (LNA) bacterial cells were determined using flow cytometry. Contrary to what is generally observed, the relative importance of HNA cells, as a percent of total cells, (%HNA, range 30-69%) was inversely related to bacterial production (range 0.15-44 ngC l(-1) h(-1)) although the negative relation was weak (log-log regression r(2) = 0.19). The %HNA as well as the mean side scatter of HNA group increased significantly with depth in the meso and bathypelagic layers. Vertical stratification played an important role in influencing the distribution and characteristics of bacterial cells especially with regard to layers located above, within or below the deep chlorophyll maximum. Within a given layer, the relationships between the flow cytometric characteristics and environmental variables such as chlorophyll-a, nutrients or bacterial production changed. Overall, the relationships between HNA and LNA cells and environmental parameters differed vertically more than longitudinally.