The attachment of bacteria to solid surfaces is influenced by substratum chemistry, but to determine the mechanistic basis of this relationship, homogeneous, well-defined substrata are required. Self-assembled monolayers (SAMs) were constructed from alkanethiols to produce a range of substrata with different exposed functional groups, i.e., methyl and hydroxyl groups and a series of mixtures of the two. Percentages of hydroxyl groups in the SAMs and substratum wettability were measured by X-ray photoelectron spectroscopy and contact angles of water and hexadecane, respectively. SAMs exhibited various substratum compositions and wettabilities, ranging from hydrophilic, hydroxyl-terminated monolayers to hydrophobic, methyl-terminated monolayers. The kinetics of attachment of an estuarine bacterium to these surfaces in a laminar flow chamber were measured over periods of 120 min. The initial rate of net adhesion, the number of cells attached after 120 min, the percentage of attached cells that adsorbed or desorbed between successive measurements, and the residence times of attached cells were quantified by phase-contrast microscopy and digital image processing. The greatest numbers of attached cells occurred on hydrophobic surfaces, because (i) the initial rates of adhesion and the mean numbers of cells that attached after 120 min increased with the methyl content of the SAM and the contact angle of water and (ii) the percentage of cells that desorbed between successive measurements (ca. 2 min) decreased with increasing substratum hydrophobicity. With all surfaces, 60 to 80% of the cells that desorbed during the 120-min exposure period had residence times of less than 10 min, suggesting that establishment of firm adhesion occurred quickly on all of the test surfaces.The consequences of bacterial adhesion to nonbiological surfaces can be beneficial or deleterious depending on the situation. Bioreactor and biofilter systems rely on the adhesion and growth of bacterial cells on support materials for effective operation (4). Also, bacterial biofilms can be utilized to promote the attachment of other organisms in the aquaculture of invertebrates (10). In marine and estuarine environments, irreversible attachment and the subsequent growth of bacteria on surfaces can affect the attachment of other organisms, often leading to the fouling of heat exchangers, ship hulls, and other synthetic surfaces (2, 18, 23). As a result, the study of bacterial adhesion and the effects of substratum surface properties has received considerable attention.Although the forces involved in bacterial adhesion are not fully understood, it is evident from numerous studies of different systems that substratum wettability is an important factor in bacterial adhesion (9, 12, 13, 26), as well as in protein adsorption (33) and the attachment of eukaryotic cells (21, 39). Many studies of bacterial adhesion and substratum wettability have utilized different materials, e.g., glass, metals, and polymers, to represent substrata with different wettabilities...