Publication Date:
2022-05-26
Description:
Author Posting. © Elsevier B.V., 2006. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Deep Sea Research Part I: Oceanographic Research Papers 53 (2006): 894-916, doi:10.1016/j.dsr.2006.01.009.
Description:
An Autonomous Microbial Sampler (AMS) is described that will obtain uncontaminated and
exogenous DNA-free microbial samples from most marine, fresh water and hydrothermal
ecosystems. Sampling with the AMS may be conducted using manned submersibles, Remotely
Operated Vehicles (ROVs), Autonomous Underwater Vehicles (AUVs), or when tethered to a
hydrowire during hydrocast operations on research vessels. The modular device consists of a
titanium nozzle for sampling in potentially hot environments (〉350°C) and fluid-handling
components for the collection of six independent filtered or unfiltered samples. An onboard
microcomputer permits sampling to be controlled by the investigator, by external devices (e.g.,
AUV computer), or by internal programming. Temperature, volume pumped and other
parameters are recorded during sampling. Complete protection of samples from microbial
contamination was observed in tests simulating deployment of the AMS in coastal seawater,
where the sampling nozzle was exposed to seawater containing 1x106 cells ml-1 of a red
pigmented tracer organism, Serratia marinorubra. Field testing of the AMS at a hydrothermal
vent field was successfully undertaken in 2000. Results of DNA destruction studies have
revealed that exposure of samples of the Eukaryote Euglena and the bacterium S. marinorubra to
0.5 N sulfuric acid at 23°C for 1 hour was sufficient to remove Polymerase Chain Reaction
(PCR) amplifiable DNA. Studies assessing the suitability of hydrogen peroxide as a sterilizing
and DNA-destroying agent showed that 20 or 30% hydrogen peroxide sterilized samples of
Serratia in 1 hr and destroyed the DNA of Serratia, in 3 hrs, but not 1 or 2 hrs. DNA AWAY™
killed Serratia and destroyed the DNA of both Serratia and the vent microbe (GB-D) of the
genus Pyrococcus in 1 hour.
Description:
This work was supported by a DOC/NOAA Small Business Innovative Research Award,
Contract No. 50-DKNA-9-90116 awarded to McLane Research Laboratories, Inc. and (via
subcontract) to the Woods Hole Oceanographic Institution. Some of the microbial testing work
was also supported by the National Science Foundation, Grant No. IBN-0131557 and the Woods
Hole Oceanographic Inst. Deep Ocean Exploration Institute Grant No. 25051131.
Keywords:
Microbiology
;
Samplers
;
DNA
;
Hydrothermal springs
;
Heat exchange
;
Aseptic
;
Pacific Ocean
Repository Name:
Woods Hole Open Access Server
Type:
Preprint
Format:
2548037 bytes
Format:
application/pdf
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