| Data Tables, Images, and Other Entities: |
| Data Table: | Units_And_Column_Descriptions
View Table Metadata |
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Short Name: | limno_bacterial_enumeration |
| Online Distribution Info: |
| Download File: |
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| Data Set Owner(s): |
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Individual: | John Priscu |
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Address: |
| Department of Biology, |
| 309 Lewis Hall, |
| Montana State University , |
| Bozeman, MT 59717 USA |
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Email Address:
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| Metadata Provider(s): |
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Organization: | McMurdo Dry Valleys LTER |
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Address: |
| Byrd Polar Research Center , |
| 108 Scott Hall, |
| 1090 Carmack Rd, |
| Columbus, OH 43210-1002 USA |
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Email Address:
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| Abstract: |
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An important part of the McMurdo Long Term Ecological Research
(LTER) project is monitoring spatial and temporal patterns, and processes that
control bacterial production in perennial ice covered lakes. This data set
quantifies bacteria concentrations at specific depths in Dry Valley lakes.
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| Keywords: |
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- lake
(theme)
- limnology
(theme)
- enumeration
(theme)
- bacteria
(theme)
- cells
(theme)
- Antarctica
(theme)
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| Additional Information: |
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| Comments |
If cells ml-1 equals zero because of the number of cells in the blank, the value is reported as zero.
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| Citations |
Hobbie, J.E., J. Daley, and S. Jasper. 1977. Use of nucleopore filters for counting bacteria by fluorescence microscopy. Applied Environmental Microbiology 33:1225-1228.
Lisle J.T. and J.C. Priscu. 2004. The occurrence of lysogenic bacteria and microbial aggregates in the lakes of the McMurdo Dry Valleys, Antarctica. Microbial Ecology. 47(4):427-439
Spigel, R.H. and J.C. Priscu. 1996. Evolution of temperature and salt structure of Lake Bonney, a chemically stratified Antarctic lake. Hydrobiologia 321:177-190.
Spigel, R.H. and J.C. Priscu. 1998. Physical limnology of the McMurdo Dry Valley lakes. In Ecosystem dynamics in a polar desert: The McMurdo Dry Valleys, Antarctica, J.C. Priscu (Ed), pp. 153-187. American Geophysical Union.
Takacs C.D. and J.C. Priscu. 1998. Bacterioplankton dynamics in the McMurdo Dry Valley lakes, Antarctica: Production and Biomass loss over four seasons. Microbial Ecology 36:239-250.
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| Notes |
Data contained in these files has been subjected to quality
control standards imposed by the investigator. The user of this data should
be aware that, while efforts have been taken to ensure that these data are
of the highest quality, there is no guarantee of perfection for the data
contained herein and the possibility of errors exists. If you encounter
questionable data, please contact the MCM LTER data manager corrected or
qualified. Thus, these data may be modified and future data will be
appended.
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| License and Usage Rights: |
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| MCM LTER data may be used freely with the following restrictions: |
The Principal Investigator be sent a notice stating
reasons for acquiring any data and a description of the
publication intentions.
The Principal Investigator of the data set be sent a
copy of the report or manuscript prior to submission and be
adequately cited in any resultant publications.
A copy of any resultant publications should be sent
to the McMurdo data manager and principal
investigator.
The end-user follow the guidelines set forth in the
LTER Network Data Access Policy, Data Access Requirements, and
General Data Use Agreement found at
http://www.mcmlter.org/data_guidelines.htm
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| Geographic Coverage: |
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Geographic Description: | Samples were collected from the East Lake Bonney, West Lake
Bonney, Lake Hoare, Lake Fryxell, and Lake Vanda limnological sampling stations, located in
the McMurdo Dry Valleys of Antarctica. |
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Bounding Coordinates:
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| West: | 162 degrees
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| East: | 163.6 degrees
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| North: | -77.2 degrees
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| South: | -77.8 degrees
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Mimimum Altitude: | 0 meter |
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Maximum Altitude: | 1000 meter |
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| Temporal Coverage: |
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Begin:
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End:
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| Maintenance: |
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Description:
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This data table was originally created by the data manager (Chris Gardner) in October of 2007.
The table was originally populated with data from the 04-05, 05-06, and 06-07 seasons that was obtained
by John Priscu's technician (Amy Chiuchiolo).
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Frequency:
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| Contact: |
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Organization: | McMurdo Dry Valleys LTER |
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Position: | Data Manager |
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Address: |
| Byrd Polar Research Center , |
| 108 Scott Hall, |
| 1090 Carmack Rd, |
| Columbus, OH 43210-1002 USA |
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Phone:
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Email Address:
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| Publisher: |
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Organization: | McMurdo Dry Valleys LTER |
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Address: |
| Byrd Polar Research Center , |
| 108 Scott Hall, |
| 1090 Carmack Rd, |
| Columbus, |
| Columbus, OH 43210-1002 USA |
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Phone:
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| Methods Info: |
| Step 1: |
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Description:
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Lake water samples are collected at specific depths with a
ive-liter Niskin bottle during normal LTER limnological sampling.
Sub-samples for bacteria enumeration are decanted into a 1 L amber
Nalgene bottle. 18 mL bacteria samples are pipetted from the amber
Nalgene bottle into new 20 ml glass scintillation vials for each depth.
Bacteria samples are preserved by adding 0.5 ml of buffered to saturation
(with sodium borate) formalin (0.2 m filtered) to each sample. Samples
are stored at 4 degrees C until preparation for counting.
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Bacteria samples are counted upon return to Montana State University.
A 0.45 um 25 mm diameter membrane filter is placed on a glass fritted filter
apparatus base and covered with a 0.2 um 25 mm diameter black polycarbonate
filter. An appropriate volume of sample (1.5-6 ml depending on the concentration
of cells in each sample) is added to a cleaned filter tower (scrubbed with Alconox,
soaked in 10% HCL, rinsed in DIW, rinsed with 95% ETOH). An appropriate
volume (0.5 ml SYBR/3 ml sample) of 0.2 µm filtered 25X SYBR Gold solution
(10,000X concentrate SYBR Gold Nucleic Acid Gel Stain in 1X TBE buffer) is added
to the sample and allowed to incubate in the dark for 15 minutes. The sample is
filtered under low vacuum (0.3 atm; 5 in Hg), and the tower rinsed with 1 ml of 0.2 um
filtered DI water when a thin layer of sample remains. The filter is placed on a glass
microscope slide containing 1 drop of Antifade solution (0.1% p-phenylenediamine
in a 1:1 (weight:volume) solution of phosphate-buffered saline and glycerol). 1 drop
of Antifade solution is placed on top of the filter before placing a cover slip on the filter.
A sample blank is prepared by following the above procedure with .2 um filtered DI water.
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Cells are counted on a Nikon Optiphot epifluorescent microscope equipped
with a DM510 filter cube (Nikon) at a final magnification of 1,000X. The total number of
cocci, rods and filaments (defined as bacteria greater than 5 um in length) are counted
in at least 10 different fields of view until at least 300 cells are counted yielding a less than
15% counting error. At least 1 digital image is taken at each depth using the Optronics
Microfire digital camera system for archiving purposes.
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cells/mL = ( (S-B) / volume ) * (filter area/field area)
where S is the average number of cells per field in the sample, B is the average number of
cells per field in the blank, filter area is the area (um2) of the filter containing bacterial cells, field area
is the area for each field counted (um2), and vol is ml of sample filtered.
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