SS2.05 Phylogenetic and Physiologic Successions in Aquatic Bacterial Communities
Date: Thursday, June 13, 2002
Location: Poster Session - VCC
WilliamsSC, Skidaway Institute of Oceanography, Savannah, USA,
Pitts, D, D, Skidaway Institute of Oceanography, Savannah, USA,
Frischer, M, E, Skidaway Institute of Oceanography, Savannah, USA,
Verity, P, G, Skidaway Institute of Oceanography, Savannah, USA,
Distinguishing Active vs. Inactive Bacterial Cells in Sediment Using the Vital Stain and Probe (VSP) Method
Distinguishing active from inactive bacteria is paramount in understanding the role bacteria play in the sediment environment. Historically, Acridine Orange and DAPI have been employed to enumerate bacteria in sediment, but this approach is unable to provide information regarding the physiological status of cells. Recently, Williams et al (1998) and Howard-Jones et al (2001) introduced the Vital Stain and Probe (VSP) method for distinguishing physiological status of individual bacterioplankton cells in aquatic samples. However, this method has not previously been employed to investigate bacteria in sediments. In an attempt to understand this complex environment, the VSP method was further modified, particularly sample preservation, hybridization time/temperature, dispersion of cells by sonication, and employment of a brighter reporter molecule. In duplicate samples that were treated simultaneously, but separately with both PI and ribosomal RNA targeted probes, the physiological status of bacterial cells was easily distinguished under wide green excitation. These observations suggest that this adapted version of the VSP method may be a powerful tool for distinguishing active and inactive bacterial cells in sediment.