http://lod.bco-dmo.org/id/dataset/684323
eng; USA
utf8
dataset
Highest level of data collection, from a common set of sensors or instrumentation, usually within the same research project
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
2017-03-14
ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data
ISO 19115-2:2009(E)
Direct counts (flow cytometry) on microbes obtained by Niskin bottle and pressure-retaining sampler from the Leggo Lander on R/V Falkor cruise FK141215 in the Challenger Deep, Mariana Trench in December 2014
2017-03-13
publication
2017-03-13
revision
Marine Biological Laboratory/Woods Hole Oceanographic Institution Library (MBLWHOI DLA)
2020-01-21
publication
https://doi.org/10.1575/1912/bco-dmo.684323.1
Douglas Bartlett
University of California-San Diego
principalInvestigator
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
publisher
Cite this dataset as: Bartlett, D. (2017) Direct counts (flow cytometry) on microbes obtained by Niskin bottle and pressure-retaining sampler from the Leggo Lander on R/V Falkor cruise FK141215 in the Challenger Deep, Mariana Trench in December 2014. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2017-03-13 [if applicable, indicate subset used]. doi:10.1575/1912/bco-dmo.684323.1 [access date]
Direct counts (flow cytometry) on microbes obtained by Niskin bottle and pressure-retaining sampler Dataset Description: <p>Direct counts (flow cytometry) on the microbes obtained in the Leggo drop 1 Niskin bottle and the Leg drop 1 pressure-retaining sampler.</p> Methods and Sampling: <p>This data set is associated with PI Douglas Bartlett (NSF OCE-1536776) and Schmidt Ocean Institute R/V Falkor cruise FK141215. The cruise occurred December 15-21, 2014 in the Challenger Deep within the territorial waters of the Federated States of Micronesia. During this cruise the Leggo lander was deployed multiple times and drops 1 and 3 recovered seawater samples that were analyzed. Additional details can be found at: <a href="https://schmidtocean.org/cruise/expanding-mariana-trench-perspectives/" target="_blank">https://schmidtocean.org/cruise/expanding-mariana-trench-perspectives/</a> and <a href="https://scripps.ucsd.edu/labs/dbartlett/contact/challenger-deep-cruise-2014/" target="_blank">https://scripps.ucsd.edu/labs/dbartlett/contact/challenger-deep-cruise-2014/</a></p>
<p><strong>Leggo Lander Drop 1:</strong><br />
Time (in Guam) deployed/recovered: December 16, 9:00/19:26.<br />
Position at deployment: 11° 21.9836 N 142° 25.9533 E, middle section of the Challenger Deep.<br />
Greatest depth of dive: approximately ~10,900 m.<br />
<em>In situ</em> temperature on seafloor: 2.6°C.<br />
Notes: This drop recovered seawater samples from about a meter off the seafloor.&nbsp;This included a 3 L&nbsp;Niskin bottle of seawater and ~ 150 mls of seawater collected in a pressure-retaining seawater sampler.&nbsp;The PRS sampler held more than 81% of the in situ pressure.&nbsp;</p>
<p><strong>Flow Cytometry:</strong><br />
Direct counts (flow cytometry) on the microbes obtained in the Leggo drop 1 Niskin bottle and the Leg drop 1 pressure-retaining sampler.&nbsp;Samples were fixed with ~1% PFA and frozen. Later samples were removed from the -80 freezer and thawed in the dark.</p>
<p>The Attune was started and a Performance Test was run with the "Performance Tracking Beads"&nbsp;to check that all lasers and filters were working, and that voltages were correct. Cleaning and decontamination was also done using the Attune Wash solution and MilliQ water. Random samples selected from Logan's samples were run using Instrument Settings in the Attune software. This allows for real-time adjustments of voltages and thresholds in the different channels to get the best resolution for that day’s run as well as allows for quantification of instrument noise for a given day.</p>
<p>Once samples were thawed, 300 uL of each sample was loaded into a 96-well U-bottom plate. 3 uL of Invitrogen Sybr-green stain (diluted to 100x in MilliQ water) was then added to each well. The plate then incubated in the dark for 30 minutes before being run. The Sybr-green stain stains any DNA within a cell which then fluoresces when passing the BL1 channel laser, allowing for counts of cells.</p>
<p>Once instrument settings were determined for the day, the plate was loaded into the NxT autosampler and the run was started. Samples were run at "Standard" sensitivity at 100 uL/min for a total volume of 250 uL. Counts were delayed for 15 seconds to avoid any noise or dilution that can occur when sample starts being sipped. Once entire plate was run, I used the Attune software to correct for noise and gate various populations within the samples.</p>
<p>Note: Leggo1 is the first drop of the Leggo Lander and the values reflect the flow cytometric cell counts from its 2 liter Niskin bottle. Leggo1_PRS_day_2 is the flow cytometric cell count obtained from the pressure retaining sampler used on the first Leggo drop, following incubation in ice water for about 36 hours after recovery.</p>
<p><strong>Colony Identification:</strong><br />
Data from the identification of bacteria cultured from the Leggo drop 1 and 3 Niskin bottles are available as a <a href="https://datadocs.bco-dmo.org/docs/bartlett/mariana_perspectives/data_docs/colony_identification.txt" target="_blank">supplemental file</a> (.txt).&nbsp;These identifications were performed using standard methods associated with PCR amplification of the 16S rRNA gene followed by dideoxy sequencing at Retrogen Inc.&nbsp;&nbsp;</p>
Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1536776 Award URL: http://www.nsf.gov/awardsearch/showAward.do?AwardNumber=1536776
completed
Douglas Bartlett
University of California-San Diego
858-534-5233
9500 Gilman Dr., MC 0210
La Jolla
CA
92093
USA
dbartlett@ucsd.edu
pointOfContact
asNeeded
Dataset Version: 1
Unknown
drop_name
cells_per_mL
ISO_DateTime_deploy
ISO_DateTime_recover
lat
lon
Niskin bottle
theme
None, User defined
sample identification
abundance
ISO_DateTime_Local
latitude
longitude
featureType
BCO-DMO Standard Parameters
Niskin bottle
Flow Cytometer
Leggo Lander
instrument
BCO-DMO Standard Instruments
FK141215
service
Deployment Activity
Challenger Deep, Mariana Trench
place
Locations
otherRestrictions
otherRestrictions
Access Constraints: none. Use Constraints: Please follow guidelines at: http://www.bco-dmo.org/terms-use Distribution liability: Under no circumstances shall BCO-DMO be liable for any direct, incidental, special, consequential, indirect, or punitive damages that result from the use of, or the inability to use, the materials in this data submission. If you are dissatisfied with any materials in this data submission your sole and exclusive remedy is to discontinue use.
Patterns of Microbial Community Structure Within and Between Hadal Environments
https://www.bco-dmo.org/project/675560
Patterns of Microbial Community Structure Within and Between Hadal Environments
<p><em>Award Abstract from NSF:</em><br />
The deepest portion of the ocean is present in ocean trenches, whose steep walls descend from approximately 4 miles down to depths that in some cases are close to 7 miles below the seawater surface. At these locations Earth's crust is recycled. Perhaps not surprisingly given their remoteness, deep ocean trenches are the least understood habitats in the ocean. The researchers participating in this project are working to characterize the microbes present in two of the deepest trenches present on Earth, both in the Pacific Ocean, the Kermadec Trench located north of New Zealand, and the Mariana Trench, located east and south of the island of Guam. Most of the Mariana Trench is located within the United States Mariana Trench Marine National Monument. Relatively little is known about the diversity and adaptations of the microorganisms in deep ocean trenches. An unknown fraction of the microbes present have descended from shallow waters above and are unlikely to participate in any nutrient cycles in the deep sea. Others are adapted to near freezing temperatures and up to pressures greater than 10e7 kilograms per square meter (16,000 pounds per square inch). These latter microbes perform important roles recycling organic matter. But who are they? This project is contributing to the training of diverse undergraduate and graduate students participating in research, additional undergraduate students learning about microbes inhabiting extreme environments in a web-based class, and additional graduate students and postdoctoral scientists participating in an advanced training course being offered in Antarctica.</p>
<p>Experiments being performed include direct counts of prokaryotes and viruses in seawater and sediments, analyses of the abundance and phylogenetic breadth of culturable heterotrophic bacteria at a range of pressures, measurements of bacterial community species diversity and richness both within and across seawater and sediment samples, as well as within and across the two trench systems, measurements of microbial activity as a function of pressure and the identification of high pressure-active cells. The data generated from these analyses are being integrated into the results of additional chemical, geological and biological measurements performed by others as a part of the National Science Foundation funded Hadal Ecosystems Studies Project. Two of the working hypotheses are that prokaryote numbers and diversity are generally positively correlated with surface productivity and proximity to the trench axis and that bacterial taxa exist which are endemic to specific trenches, present in multiple trenches and more widely distributed in deep-sea environments.</p>
Mariana Perspectives
largerWorkCitation
project
eng; USA
oceans
Challenger Deep, Mariana Trench
142.432555
142.432555
11.36639
11.36639
2014-12-16
2014-12-16
Challenger Deep, Mariana Trench
0
BCO-DMO catalogue of parameters from Direct counts (flow cytometry) on microbes obtained by Niskin bottle and pressure-retaining sampler from the Leggo Lander on R/V Falkor cruise FK141215 in the Challenger Deep, Mariana Trench in December 2014
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
http://lod.bco-dmo.org/id/dataset-parameter/684333.rdf
Name: drop_name
Units: unitless
Description: Name of the lander drop
http://lod.bco-dmo.org/id/dataset-parameter/684334.rdf
Name: cells_per_mL
Units: cells/mL
Description: Number of cells per milliliter (mL)
http://lod.bco-dmo.org/id/dataset-parameter/684335.rdf
Name: ISO_DateTime_deploy
Units: unitless
Description: Date and time (local Guam time zone) of lander deployment; formatted to ISO 8601 standard.
http://lod.bco-dmo.org/id/dataset-parameter/684336.rdf
Name: ISO_DateTime_recover
Units: unitless
Description: Date and time (local Guam time zone) of lander recovery; formatted to ISO 8601 standard.
http://lod.bco-dmo.org/id/dataset-parameter/684337.rdf
Name: lat
Units: decimal degrees
Description: Latitude of lander deployment
http://lod.bco-dmo.org/id/dataset-parameter/684338.rdf
Name: lon
Units: decimal degrees
Description: Longitude of lander deployment
GB/NERC/BODC > British Oceanographic Data Centre, Natural Environment Research Council, United Kingdom
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
240
https://darchive.mblwhoilibrary.org/bitstream/1912/25213/1/dataset-684323_flow-cytometry__v1.tsv
download
https://doi.org/10.1575/1912/bco-dmo.684323.1
download
onLine
dataset
<p>This data set is associated with PI Douglas Bartlett (NSF OCE-1536776) and Schmidt Ocean Institute R/V Falkor cruise FK141215. The cruise occurred December 15-21, 2014 in the Challenger Deep within the territorial waters of the Federated States of Micronesia. During this cruise the Leggo lander was deployed multiple times and drops 1 and 3 recovered seawater samples that were analyzed. Additional details can be found at: <a href="https://schmidtocean.org/cruise/expanding-mariana-trench-perspectives/" target="_blank">https://schmidtocean.org/cruise/expanding-mariana-trench-perspectives/</a> and <a href="https://scripps.ucsd.edu/labs/dbartlett/contact/challenger-deep-cruise-2014/" target="_blank">https://scripps.ucsd.edu/labs/dbartlett/contact/challenger-deep-cruise-2014/</a></p>
<p><strong>Leggo Lander Drop 1:</strong><br />
Time (in Guam) deployed/recovered: December 16, 9:00/19:26.<br />
Position at deployment: 11° 21.9836 N 142° 25.9533 E, middle section of the Challenger Deep.<br />
Greatest depth of dive: approximately ~10,900 m.<br />
<em>In situ</em> temperature on seafloor: 2.6°C.<br />
Notes: This drop recovered seawater samples from about a meter off the seafloor.&nbsp;This included a 3 L&nbsp;Niskin bottle of seawater and ~ 150 mls of seawater collected in a pressure-retaining seawater sampler.&nbsp;The PRS sampler held more than 81% of the in situ pressure.&nbsp;</p>
<p><strong>Flow Cytometry:</strong><br />
Direct counts (flow cytometry) on the microbes obtained in the Leggo drop 1 Niskin bottle and the Leg drop 1 pressure-retaining sampler.&nbsp;Samples were fixed with ~1% PFA and frozen. Later samples were removed from the -80 freezer and thawed in the dark.</p>
<p>The Attune was started and a Performance Test was run with the "Performance Tracking Beads"&nbsp;to check that all lasers and filters were working, and that voltages were correct. Cleaning and decontamination was also done using the Attune Wash solution and MilliQ water. Random samples selected from Logan's samples were run using Instrument Settings in the Attune software. This allows for real-time adjustments of voltages and thresholds in the different channels to get the best resolution for that day’s run as well as allows for quantification of instrument noise for a given day.</p>
<p>Once samples were thawed, 300 uL of each sample was loaded into a 96-well U-bottom plate. 3 uL of Invitrogen Sybr-green stain (diluted to 100x in MilliQ water) was then added to each well. The plate then incubated in the dark for 30 minutes before being run. The Sybr-green stain stains any DNA within a cell which then fluoresces when passing the BL1 channel laser, allowing for counts of cells.</p>
<p>Once instrument settings were determined for the day, the plate was loaded into the NxT autosampler and the run was started. Samples were run at "Standard" sensitivity at 100 uL/min for a total volume of 250 uL. Counts were delayed for 15 seconds to avoid any noise or dilution that can occur when sample starts being sipped. Once entire plate was run, I used the Attune software to correct for noise and gate various populations within the samples.</p>
<p>Note: Leggo1 is the first drop of the Leggo Lander and the values reflect the flow cytometric cell counts from its 2 liter Niskin bottle. Leggo1_PRS_day_2 is the flow cytometric cell count obtained from the pressure retaining sampler used on the first Leggo drop, following incubation in ice water for about 36 hours after recovery.</p>
<p><strong>Colony Identification:</strong><br />
Data from the identification of bacteria cultured from the Leggo drop 1 and 3 Niskin bottles are available as a <a href="https://datadocs.bco-dmo.org/docs/bartlett/mariana_perspectives/data_docs/colony_identification.txt" target="_blank">supplemental file</a> (.txt).&nbsp;These identifications were performed using standard methods associated with PCR amplification of the 16S rRNA gene followed by dideoxy sequencing at Retrogen Inc.&nbsp;&nbsp;</p>
Specified by the Principal Investigator(s)
<p>BCO-DMO Processing:<br />
-modified parameter names to conform with BCO-DMO naming conventions;<br />
-replaced spaces with underscores;<br />
-replaced '-' with 'nd' (no data);<br />
-added dates, times, locations from metadata form.</p>
Specified by the Principal Investigator(s)
asNeeded
7.x-1.1
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
Niskin bottle
Niskin bottle
PI Supplied Instrument Name: Niskin bottle Instrument Name: Niskin bottle Instrument Short Name:Niskin bottle Instrument Description: A Niskin bottle (a next generation water sampler based on the Nansen bottle) is a cylindrical, non-metallic water collection device with stoppers at both ends. The bottles can be attached individually on a hydrowire or deployed in 12, 24, or 36 bottle Rosette systems mounted on a frame and combined with a CTD. Niskin bottles are used to collect discrete water samples for a range of measurements including pigments, nutrients, plankton, etc. Community Standard Description: http://vocab.nerc.ac.uk/collection/L22/current/TOOL0412/
PI Supplied Instrument Name: Instrument Name: Flow Cytometer Instrument Short Name:Flow Cytometer Instrument Description: Flow cytometers (FC or FCM) are automated instruments that quantitate properties of single cells, one cell at a time. They can measure cell size, cell granularity, the amounts of cell components such as total DNA, newly synthesized DNA, gene expression as the amount messenger RNA for a particular gene, amounts of specific surface receptors, amounts of intracellular proteins, or transient signalling events in living cells.
(from: http://www.bio.umass.edu/micro/immunology/facs542/facswhat.htm) Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB37/
PI Supplied Instrument Name: Instrument Name: Leggo Lander Instrument Short Name: Instrument Description: The "Leggo Lander" is a lander system that primarily relies on syntactic foam for buoyancy and uses iridium GPS, radio signal, strobe light and flag for surface recovery, and acoustics for underwater monitoring and instrument control. The lander has a timer with 5 control settings for various operations. It routinely measures pressure (depth) throughout its dive and temperature on the seafloor. The lander payloads include a pressure-retaining seawater sampler plus 2 liter Niskin bottle, and a camera/battery/light system that also includes a 30 liter Niskin bottle and a sea cucumber trap. With the camera payload it travels down or up the water column at about 39 meters per minute (~ 4.5 hours for a descent to the Challenger Deep at ~10,920 m).
(Description obtained from the R/V Falkor FK141215 post-cruise report (PDF))
Cruise: FK141215
FK141215
R/V Falkor
Community Standard Description
International Council for the Exploration of the Sea
R/V Falkor
vessel
FK141215
Douglas Bartlett
University of California-San Diego
http://dmoserv3.whoi.edu/data_docs/Mariana_Perspectives/Bartlett-final-FK141215-cruise-report.pdf
Report describing FK141215
R/V Falkor
Community Standard Description
International Council for the Exploration of the Sea
R/V Falkor
vessel