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Enumeration Of Sandy Sediment Bacteria: Search For Optimal Protocol, Slava Epstein, Jeffrey Rossel
Enumeration Of Sandy Sediment Bacteria: Search For Optimal Protocol, Slava Epstein, Jeffrey Rossel
Slava Epstein
We examined and compared a variety of existing protocols for enumeration of bacteria from marine sandy sediments. The focus was on how to dislodge bacteria from sediment particles; a commercial blender, an ultrasonic cleaner, and an ultrasonic cell disrupter were tested. The ultrasonic cell disrupter was found to be the most efficient device for bacterial dislodgment. With a 5 mm microtip vibrating at 109 μm amplitude and 20 kHz, the optimal sonication time of small (≤ 0.5 cm³) samples was 180 s. Having identified the optimal dislodgment treatment, we went through other steps of sediment bacteria enumeration (use of surfactants, …
Enumeration Of Sandy Sediment Bacteria: Are The Counts Quantitative Or Relative?, Slava S. Epstein, D. Alexander, K. Cosman, A. Dompé, S. Gallagher, J. Jarsobski, E. Laning, R. Martinez, G. Panasik, C. Peluso, R. Runde, E. Timmer
Enumeration Of Sandy Sediment Bacteria: Are The Counts Quantitative Or Relative?, Slava S. Epstein, D. Alexander, K. Cosman, A. Dompé, S. Gallagher, J. Jarsobski, E. Laning, R. Martinez, G. Panasik, C. Peluso, R. Runde, E. Timmer
Slava Epstein
Several tests were carried out to enable evaluation of the precision with which sandy sediment bacteria could be enumerated. This represented the first attempt to place direct epifluorescence counts of benthic bacteria on a quantitative, rather than relative, ground. The tests combined in situ radioisotope ([³H], [¹⁴C]) labeling of sediment bacteria, bacterial dislodgment by ultrasonic treatment, and bacterial enumeration via fluorescent staining. The results provided direct and indirect evidence that the employed protocol for bacterial enumeration accounted for 88 to 98% of all bacteria present in sediments. The identified approach thus allowed for a rather complete quantification of sediment bacteria. …