The Hawaiian Archipelago is a "biodiversity hotspot" where significant endemism among eukaryotes has evolved through geographic isolation and local topography. To address the absence of corresponding region-wide data on Hawaii's microbiota, we compiled the first 16S SSU rDNA clone libraries and cultivated bacteria from five Hawaiian lakes, an anchialine pool, and the Lō'ihi submarine volcano. These sites offer diverse niches over approximately 5000 m elevation and approximately 1150 nautical miles. Each site hosted a distinct prokaryotic community dominated by Bacteria. Cloned sequences fell into 158 groups from 18 Bacteria phyla, while seven were unassigned and two belonged in the Euryarchaeota. Only seven operational taxonomic units (each OTU comprised sequences that shared > or =97% sequence identity) occurred in more than one site. Pure bacterial cultures from all sites fell into 155 groups (each group comprised pure cultures that shared > or =97% 16S SSU rDNA sequence identity) from 10 Bacteria phyla; 15 Proteobacteria and Firmicutes were cultivated from more than one site. One hundred OTUs (60%) and 52 (33.3%) cultures shared <97% 16S SSU rDNA sequence identity with published sequences. Community structure reflected habitat chemistry; most delta-Proteobacteria occurred in anoxic and sulfidic waters of one lake, while beta-Proteobacteria were cultivated exclusively from fresh or brackish waters. Novel sequences that affiliate with an Antarctic-specific clade of Deinococci, and Candidate Divisions TM7 and BRC1, extend the geographic ranges of these phyla. Globally and locally remote, as well as physically and chemically diverse, Hawaiian aquatic habitats provide unique niches for the evolution of novel communities and microorganisms.

Absfmct -Hydrocarbon seep monitoring is a area applicable for new advances in mass spectrometry. Recent developments are allowing unprecedented levels of trace contaminant measurement in the deep-ocean. With funding from the U.S. National Science Foundation (NSF), our engineering design team constructed a new mass spectrometer-based in situ analysis system for work in the deep ocean environment over prolonged deployment periods.Our design goals were a depth capability of up to 4,000 m water depth (400 bars hydrostatic pressure) and autonomous operation for periods of up to sir months to a year, depending upon type of exteFa1 battery system used or other deployment circumstances, e.g., availability of a power cable or fuel cell power source. We chose a membrane introduction mass spectrometry (MIMS) sampling approach, which allows for dissolved gases and volatile organics introduction into the MS vacuum system. The MIMS approach and the hydrophobic, silicon-coated membrane chosen both draw upon our previous experience with this technology in the deep ocean. The membrane has been tested to 400 bars in a series of long-term hydrostatic pressure tests, which extends the 200-bar working depth rating of this membrane by a factor of 2.

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New developments in instrumentation for ocean environmental engineering are allowing unprecedented levels of trace contaminant measurement in the deep ocean. With funding from the U.S. National Science Foundation (NSF), our engineering design team constructed a new mass spectrometerbased in situ analysis system for work in the deep ocean environment over prolonged deployment periods. Our design goals were a depth capability of up to 4,000 m water depth (400 bars hydrostatic pressure) and autonomous operation for periods of up to six months to a year, depending upon the type of external battery system used or other deployment circumstances, e.g., availability of a power cable or fuel cell power source. We chose a membrane introduction mass spectrometry (MIMS) sampling approach, which allows for dissolved gases and volatile organics introduction into the mass spectrometer vacuum system. The MIMS approach and the hydrophobic, silicon-coated membrane chosen both draw upon our previous experience with this technology in the deep ocean. The membrane has been tested to 400 bars in a series of longterm hydrostatic pressure tests, which extend the 200-bar working depth rating of this membrane by a factor of 2. Longterm deployment capability of the moderately powered, approximately 100 W system, was accomplished by power management of the embedded computer system and custom electronics with Windows-based and custom software now fully-developed and bench tested.The entire system fits within a 6.5-inch outside diameter pressure housing that is approximately five feet long. It consists of a 1 to 200 amu range quadrupole mass spectrometer equipped with Faraday and electron multiplier detectors, compact turbo-molecular and backing diaphragm vacuum pumps, internal rechargeable batteries, and internal waste vacuum chamber. Sample routing past the MIMS is accomplished by computer-controlled solenoid valves. We designed the pressure housings of both 6AL4V and type 2 titanium alloys that are rated to working depths of >4,000 m and are essentially corrosion proof over long-term deployments. We designed and integrated a fail-safe valving system for both rapid response to high-pressure MIMS failure and a pressure-switch circuit and high-pressure solenoid valve to detect and protect against slow leaks of the MIMS. To route sample waters to the MIMS-based instrument, we also designed and built a rugged plastic plenum that couples to the face of the sampler head, the latter of which consists of the MIMS inlet and a full-ocean rated thermister temperature probe with an operational range from -5 to 50°C. These instrumentation innovations will be described in the paper.