Use of Sonication for Enhanced Sampling of Attached Microbes from Groundwater Systems

The vast majority of microorganisms in aquifers live as biofilms on sediment surfaces, which presents significant challenges for sampling as only the suspended microbes will be sampled through normal pumping. The use of a down-well low frequency sonicator has been suggested as a method of detaching microbes from the biofilm and allowing rapid sampling of this community. We developed a portable, easy to use, low-frequency electric sonicator and evaluated its performance for a range of well depths (tested up to 42 m below ground level) and casing types. Three sonicators were characterized in laboratory experiments using a 1 m long tank filled with pea gravel. These included a commercially available pneumatic sonicator, a rotating flexible shaft sonicator, and the prototype electric sonicator. The electric sonicator detached between 56 and 74% of microbes grown on gravel-containing biobags at distances ranging between 2 and 50 cm from the sonicator. The field testing comprises of a total of 55 sampling events from 48 wells located in 4 regions throughout New Zealand. Pre- and post-sonication samples showed an average 33 times increase in bacterial counts. Microbial sequence data showed that the same classes are present in pre- and post-sonicated samples and only slight differences were seen in the proportions present. The sampling process was rapid and the significant increases in bacterial counts mean that microbial samples can be quickly obtained from wells, which permits more detailed analysis than previously possible.