Microbiological investigations utilizing molecular genetic approaches to characterize microbial communities can require large volume water samples, tens to hundreds of liters. The requirement for large volume samples can be especially challenging in deep-sea hydrothermal vent environments of the oceanic ridge system. By and large studies of these environments rely on deep submergence vehicles. However collection of large volume (>100L) water samples adjacent to the benthos is not feasible due to weight considerations. To address the technical difficulty of collecting large volume water samples from hydrothermal diffuse flow environments, a semi-autonomous large-volume water sampler (LVWS) was designed. The LVWS is capable of reliably collecting and bringing to the surface 120L water samples from diffuse flow environments. Microscopy, molecular genetic and chemical analyses of water samples taken from 9°N East Pacific Rise are shown to demonstrate the utility of the LVWS for studies of near-benthos environments. To our knowledge this is the first report of virioplankton abundance within diffuse-flow waters of a deep-sea hydrothermal vent environment. Because of its simple design and relatively low cost, the LVWS should be applicable to a variety of studies which require large-volume water samples collected immediately adjacent to the benthos.