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ARCTIC LTER
LAKES FIELD SEASONPROTOCOLS
Summer 2002
Prepared ByM. Bahr N. Bettez A. Hershey J. Hobbie G. Kipphut G. Kling J. Laundre M. McDonald M. Miller J. O’Brien P. Rublee TABLE OF CONTENTS I. CALIBRATION OF FIELD EQUIPMENT................................................................................................................. 4 A. Hydrolab multiprobe.............................................................................................................................................. 4 B. Licor light sensor.................................................................................................................................................... 2 II. Calibration of Laboratory Equipment................................................................................................ III. FIELD SAMPLING BOTTLES................................................................................................................................. 2 IV. FIELD SAMPLING BOTTLE PREPARATION...................................................................................................... 2 V. PREPARING FOR FIELD SAMPLING..................................................................................................................... 3 A. Prior Preparation..................................................................................................................................................... 3 B. Sampling Strategies................................................................................................................................................ 3 C. Field Checklist for sampling of inner lakes......................................................................................................... 4 D. Checklist of gear required for biological sampling of outer lakes and surveys............................................ 5 E. Field Checklist for full sampling of outer lakes and surveys............................................................................ 6 F. Checklist of gear to remain in packs at all times................................................................................................. 6 G. Personal gear required for sampling.................................................................................................................... 6
VI.
FIELD SAMPLING....................................................................................................................................................
6
A. Oxygen, conductivity, temperature..................................................................................................................... 7 B. Secchi depth............................................................................................................................................................ 7 C. Integrated Phytoplankton sampling.................................................................................................................... 7 D. Light......................................................................................................................................................................... 7 E. Water sample Collection for Primary production, chlorophyll, and nutrients............................................... 7 F. Field Filtering for nutrients.................................................................................................................................... 7 G. Collection of survey Water Sample..................................................................................................................... 8 H. Collection of a Grab sample.................................................................................................................................. 8 I. Zooplankton Sampling........................................................................................................................................... .8 J. Microplankton Sampling........................................................................................................................................ 9 K. Benthos Sampling................................................................................................................................................ 10 L. Fish Sampling........................................................................................................................................................ 11
VII. BACTERIA SAMPLING AND MEASUREMENT............................................................................................. 12 A. Water Collection.................................................................................................................................................. 12 B. Bacteria Counts..................................................................................................................................................... 13 C. Bacteria Production.............................................................................................................................................. 13
VIII. PROCESSING SAMPLES IN LABORATORY.................................................................................................. 13 VIX. LABORATORY ANALYSES.............................................................................................................................. 15 A. Primary Production.............................................................................................................................................. 15 B. Alkalinity (Gran Titration)................................................................................................................................... 16 C. Winkler Oxygen Titration.................................................................................................................................... 16
X. DATA ENTRY.......................................................................................................................................................... 21 A. Surveyor download............................................................................................................................................. 15 B. Licor download..................................................................................................................................................... 16 C. Winkler Oxygen Titration.................................................................................................................................... 16
XI. Arctic LTER Procedures for on-line Data Management....................................................... 22 XII. PROTOCOL FOR ENTERING DATA FILES INTO THE LTER DATABASE............................................... 23 XIII. INSTRUCTIONS FOR COMPLETION OF DOCUMENTATION FILE......................................................... 23 XIV. GLOBAL POSITIONING SYSTEM INSTRUCTIONS..................................................................................... 25 XV. USE LTER MOTOR BOATS (Toolik Lake Map)............................................................................................... 30 XVI. END OF SEASON................................................................................................................................................. 31 XVII. ADDRESS BOOK................................................................................................................................................ 32 XVIII. PROTOCOL REFERENCES............................................................................................................................... 35 XIX. NOTES................................................................................................................................................................... 36 ARCTIC LTER LAKES PROTOCOLS I. CALIBRATION OF FIELD EQUIPMENT
A. The Hydrolab unit consists of the H2O multi-parameter water quality data transmitter and the Surveyor 3 Multi-parameter water quality logging system. The H20 is capable of measuring Temperature, specific conductance, pH, and Dissolved Oxygen. Before the field season all routine maintenance on sensors should be performed. This includes cleaning specific conductance and pH sensors, service to the reference electrode and changing the DO sensor membrane. The specific conductance calibration is stable for six months and should be calibrated at the start of the field season. The dissolved oxygen and pH are stable for 1 month but are calibrated weekly while at Toolik. All sensors are inspected weekly for wear and cleanliness. This is usually done on Friday since most require over night equilibration after maintenance. The maintenance scheduled for the H20 is: Specific conductance sensor clean and calibrate monthly pH glass electrode inspect and calibrate weekly Reference electrode replace electrolyte weekly and inspect to make sure it passes electrolyte freely. Dissolved Oxygen membrane: inspect and calibrate weekly and replace membrane and electrolyte monthly. Depth sensor inspect monthly clean if needed. The surveyor has internal lithium batteries that require replacement every two years. If the surveyor fails to keep track of Dates the batteries need to be replaced. The manuals for these pieces of equipment are thorough in explaining use and calibration. Calibration should be done each Saturday before the following week’s sampling run. The surveyor unit must be charged to between 14.4 and 16 volts. An over night charge before sampling is usually adequate. Following is a brief synopsis of the oxygen calibration.. 1. Get the barometric pressure either from the hand held barometer or data logger. To read the barometric pressure off the hand held unit look at the window in the 12 o’clock position to see what color the white arrow above the km is pointing to. This is your height above sea level and indicates the scale you should read of off (my guess is it will be red or maybe yellow). After this tap the glass lightly then read the BP off the correct scale. 2. Plug appropriate end of cable into right side (back) of surveyor 3 (Transmitter). Plug other end into H20 datasond unit making sure the plugs are aligned correctly. This involves aligning the dots on the plug end of the cable with the largest prong on the male plug end of the H2O datasond unit (Hydrolab multiprobe). Some force is involved but not too much, so be patient and make sure it is correct before applying too much force. 3. Turn the surveyor 3 on [On / Off] The response time varies for each parameter but all are usually less than one minute. 4. Remove cup containing DI water covering sensors. 5. Hit blue [calibrate] key followed by the [% / S / DO] key. The screen will show the following % O. Highlight the % using the arrow keys followed by the enter key. 6. You will then be prompted for the barometric pressure, enter it followed by the [enter] key. You will see the message Sending….. (this is the surveyor 3 sending the calibration to the H20 datasond unit). The other possible message is Out of tolerance no calibration saved. If this appears make sure the Barometric pressure is correct, other possible reasons are, a damaged membrane, or low battery. If the membrane is intact and battery charged (checked by hitting the screen key, charge should appear in the lower right and should be above 15.00 V&
B. Licor light meter: The batteries should be checked weekly on the Licor. There are no means for calibrating this meter at Toolik so all calibration should be done prior to the field season. LI-COR recommends that the probes be calibrated every 2 years. Sensor must be connected to correct connection ports since each quantum sensor has its own multiplier.
The LI 1400 has two light sensors the deck (Q25190 ) and the submersible (UWQ 4879). Each sensor has a unique calibration multiplier assigned to it by Licor. Both calibration multipliers are stored in a set up file on computers used to download files as well as on the disk containing the software. Log routines for either logging (logroutinesetup.L14) and no logging set up (licorsetup.L14) are both stored on the diskette containing the Licor manual. The calibration constants for each of the sensors are: Underwater (UWQ 4879): -242.13 Deck (Q 25190): -214.59
To Use the Licor for instantaneous measurements 1. plug each sensor into its correct current channel (I1 for the Deck and I2 for the sub. 2. Turn Logger on [on/off] 3. hit [view] scroll between {log data} and new data using the arrow key [à] 4. Select New data by hitting enter [enter] This menu is an array of 4 lines each with 7 choices (see diagram). Each of the four lines can display any of the seven choices. You will be on the first line of the 4. Move among the 7 choices using the left or right arrow keys. Your choices are: Data and time (YYYYMMDD) (HHMMSS) Memory (total avail) (amount used)
Channel 1 (light data) DEC
Channel 1 log status [on or off]
Channel 2 (light data) DEC
Channel 2 log status [on or off] Battery voltage 5. When I1 I DEC (or Q25190) appears. Hit down arrow to move to next line in menu. 6. Scroll left or right until I2 I Sub (or UWQ 4879) appears 7. If Q 25190 and UWQ appear hit the [view] key to toggle to Dec and sub ID’s. The data is the same but it is easier to keep sensors straight using Dec and sub. 8. When ready to store data simply press enter key. Remember that no ID are stored with data so you must remember for what and where you are storing data. This is why it is often easier to also write the numbers down as you collect the data in this mode.
To set the LI1400 up for logging 1. plug each sensor into its correct current channel (I1 for the Deck and I2 for the sub. 2. Turn Logger on [on/off] 3. Hit [set up] One of 7 choices will appear in the menu. Scroll though using the left of right arrow keys [à] until logging appears 4. hit enter[enter] 5. Toggle between choices on and off using arrow keys 6. LI1400 is now logging and will continue to do so until logging is turned off. If you fail to turn it off the batteries will die in about three days. 7. When logging is done repeat steps 2-6 and toggle logging to off. 8. Note. As with most limnological equipment the Licor is not waterproof and when left out to log it should be kept in a zip lock.
Downloading Licor LI1400 data logger 1. Using Beige null modem cable, plug 9 pin (female) to male 9 pin on bottom of LI1400. Plug other 9 pin female to com 1 on laptop (other 9 pin male). 2. Open Licor download software. If not installed install using enclosed disks. 3. On top drop down menu click on Remote then click on connect. You can also hit [alt] R [Return]. You are now connected to LI 1400. 4. To down load data: Click on remote menu again followed by receive data. 5. You will be prompted for a range. I usually select all [return] 6. You will then be asked to pick a directory and name the file. 7. Hit save and file will be saved to directory. 8. The file is a delimited text file, which is readable by, excel. 9. Before clearing the database open the file and make sure the download was a success. Once it is and the data is backed up clear the database for the next log. 10. Click on remote and select clear database or [Alt] R [Shift] C. 11. You then have the choice to clear all or portions of the database. You will then be warned again that you are about to clear the database. If this is ok go for it.
II. Calibration of the Turner Designs 10-AU for Chlorophyll measurements.
If you are not sure that the correct
filter set and lamp are installed
for measurement of chlorophyll you
must open the machine and check. However, once this is done you will be
required to recalibrate the
fluorometer.
The Arctic LTER has used Turner Designs optical kit (10-037R), which is
for the measurement of invivo and
extractive measurement of
chlorophyll. This kit is used for the traditional in
vivo measurement of chlorophyll
(Lorenzen) and invivo /extractive
acidification methods, including
Strickland and parsons, standard
methods for water and wastewater and
EPA 445.
The Turner Designs chlorophyll optical kit (10-037R)
contains:
1.
10-AU-600 Red
sensitive photomultiplier tube
2.
10-AU 45
daylight white lamp (F4T5D),
3.
10-050R Round
Excitation filter (340-500 nm)
4.
10-051R Round
emission filter >665 nm
5.
10-032 Square
reference filter 400-700 (1ND)
In order to access any of the
screens you must first get to the
home screen. This is accomplished by hitting enter
[ENT]
Calibration is begun from screen 2
<2> on the main menu. The basic operating parameters and level
of the 10-AU must be set prior to
your calibrating the machine.
1.
Operational
parameters
1.2. Home display
Readout
conc
Units
None
1.4. Output
Full scale
V:2V
Zero
0
2. Calibration
2.1. Blanking
Subtract blank
NO
2.2. Standard SOL
Standard conc.
Concentration of solid standard
(54.988)
2.4.
1.
Display range
Yes
2.
Set range High
3.
Set range
control
Man
Also prior to calibration the range
for the instrument must be set.
There is a trade off between range
and sensitivity.
The larger the range that an instrument is set to measure the less
sensitive it becomes.
Sensitivity can be thought of as the
degree of precision.
Because chlorophyll is only linear up to 250 ug/l we have chosen that as
our upper limit, so our range is
0-250.
There are three ranges on the 10-AU each 1 order of magnitude apart.
Low 0-2.5, Med 2.5-25, and High
25-250.
To set the range of the 10-AU place
a standard in the cuvette that is
approximately 20% of the maximum
concentration that you wish to read
(~ 50 ug).
Go to screen 2.3 and set the span to between 30 and 50 using the up and
down arrow, remember that the higher
the span the higher the sensitivity. Next adjust the FS% so that the high is
250, med is 25 and low is 2.5 using
the sensitivity adjustment knob,
which is located to the right of the
keypad.
After each adjustment of the sensitivity knob you will need to wait 1-8
seconds for the machine to stabilize
again.
Turning the knob clockwise will make the machine more sensitive and
reduce the range while turning it
counter clockwise will do the
opposite. Once the high reading has stabilized
around 250 hit enter.
Note that you can also adjust the
span up or down for fine scale
adjustment of the sensitivity
(increasing the span will decrease
the range).
When this is done go to screen 3
(Diagnostics) and with the high
standard still in the cuvette holder
record the following, when finished
repeat for the low standard.
PWR level
Chopper RPM
High volt
Fluor readout
PM (photo multiplier) signal output
Cal STD value
Blank
FS: % of
Span
The fluorometer gives readings in
total ug chlorophyll ml-1
Acetone.
This must be converted to ug /L
water (seston) or ug/cm2 of rock
surface (rock scrubs). To calculate concentration from your
fluorometer readings you will need
to know volume filtered and volume
of acetone used to extract your
sample.
If you are reading sestonic samples
the equation would be ug total
chlorophyll liter -1 = (reading) x
(liters acetone / liters filtered)
If you are reading scrub samples the
equation would be ug total
chlorophyll liter -1 = (reading) x
(liters acetone / liters of scrub
sample filtered) x (L of water in
scrub sample/cm2 of rock area
scrubbed)
III. FIELD SAMPLING BOTTLES
A. Water samples are stored in 1 liter amber bottles and brought back to the lab for processing. Chlorophyll (47 mm GF/C) Particulate N and C, Particulate P, (25mm GF/C Alkalinity, (60 ml HDPE round) Cations, (60 ml HDPE round) Anions, (30 ml HDPE round)
C. Samples to be analyzed for Phosphate and ammonium by hand using wet chemistry are collected in 60 ml HDPE amber bottles. These bottles are labeled and dedicated for each lake. D. Integrated Phytoplankton samples are collected in 125 ml bottles. Preserved in the lab concentrated and shipped to Hedy Kling in Aug. E.
Zooplankton samples are collected in
250 ml
wide mouth bottles preserved with
Buffered Formalin and shipped to
John O’Brien in
F. Microplankton are collected in 60 ml square polypropylene bottles. G. Total dissolved Nitrogen are collected in clear round 60ml trace metal clean LDPE bottles, preserved with 100 ul/ 60ml 6N Ultrex HCL, and frozen at Toolik for shipment to MBL. F. Total dissolved Phosphorus are collected in clear round 60 ml HDPE, preserved with 100 ul/ 60ml 6N Ultrex HCL, and stored at 4 C at Toolik and shipped to MBL.
IV. FIELD SAMPLING BOTTLE PREPARATION
A. The 1 liter field sampling bottles should be rinsed with DDW after sample water is processed. B. Bottles for inorganic nutrient sampling should be soaked in the acid bath (10% HCl) for at least a few hours and then rinsed 3 times with DI water and filled DI until samples are taken. C. Water for Primary productivity is collected in a 500 ml amber bottle and brought back to the lab for processing.
V. PREPARING FOR FIELD SAMPLING
There will be a calendar in the lab indicating the sampling schedule for this summer. A. Prior to sampling (the night before) make sure all bottles and Petri dishes are clean and labeled. Indicate on label the lake, depth, date sampled. USE TAPE!! Writing on the bottle only results in a lot of blank bottles when it comes time for the samples to be analyzed. Each lake is color-coded to facilitate later analyses. Fill 15 ml centrifuge tubes with 10 ml acetone (for chlorophyll a analyses). Fill out Filter and Chlorophyll books and label Nutrient and chlorophyll tubes with corresponding numbers. B. We are using 2 sampling strategies. Biological samples and measurements will be taken every time while full sampling (biological and nutrient samples and measurements) will occur only when indicated.
The sampling depths for lakes are as follows: Toolik: 0,1,3,5,8,12,16 N-1: 0,1,3,5,8,12 S6: 0,1,3,5, S7: 0,1,3 N2 R: 0,1,3,5,6 N2T: 0,1,3,5,8 I 7: 0,1,3,5,8,12 The remainder are sampled at epilimnion, metalimnion and hypolimnion
Biological
sampling includes:
1. Zooplankton and phytoplankton samples are collected on each sampling trip. 2. Microplankton samples are taken at every lake and depth sampled (except I series inlets and outlets) 3. Primary Production samples are taken at every lake and depth sampled. 4. Chlorophyll a samples are taken are taken at every lake and depth sampled. 5. pH, temperature, conductivity, oxygen, measured at every meter on every lake and light (0, .1, .2, .5, 1 and every meter to the bottom). 6. Secchi depth is measured on every lake 7. Alkalinity samples are taken at every lake and depth sampled. In Toolik samples are taken from each sampled depth for the first sample date only. For the remainder of the summer only epilimnion, metalimnion and hypolimnion samples are taken.
Full sampling includes the following in addition to Biological sampling:
1. Cation and anion samples are taken at every lake and depth sampled. In Toolik samples are taken from each sampled depth for the first sample date only. For the remainder of the summer only epilimnion, metalimnion and hypolimnion samples are taken. 2. Inorganic nutrient samples (NH-4, NO-3, PO-4) are collected from each depth sampled in each lake (or inlet) . 3. TDN and TDP samples are collected from each depth sampled in each lake (or inlet) . 4. Particulate (N, C, P) samples are collected from each depth sampled in each lake (or inlet) . 5. DOC samples are collected from each depth sampled in Toolik, N1, N2, S6, S7, and the I series. 6. Bacteria samples are collected from each depth sampled in Toolik.
C. FIELD CHECKLIST SAMPLING OF INNER LAKES 1. Zooplankton net 2. Squeeze bottle for rinsing microplankton and zooplankton nets 3. Integrated Phytoplankton Sampler (meter-marked Tygon tubing) 4. 3 l pitcher and 20 um nitex for microplankton concentration 5. Bottles for collection of primary productivity samples (500 ml amber) 7. Van Dorn or water pump/pump tubing. 8. Hydrolab (H2O) and Surveyor 3, including cable and stirrer 9. Secchi disk 10. Licor light meter, including cable 11. Field notebook (Rite-in-Rain), pencil, Sharpie 12. Nalgene filter rig, hand-held vacuum pump, and .45 micron Millipore filters, forceps 13. 1 liter amber bottle for collection of water to return to lab from which Cations, anions, alkalinity, PN,PC, PP DOC, and chlorophyll a will be taken. 14. Bag containing sample bottles for each sample depth:.
17. Sample bottles for zooplankton and integrated phytoplankton
D. CHECKLIST OF GEAR REQUIRED FOR BIOLOGICAL SAMPLING OF OUTER LAKES AND SURVEYS 1. Zooplankton net 2. Squeeze bottle for rinsing microplankton and zooplankton nets 3. Integrated Phytoplankton Sampler (meter-marked Tygon tubing) 4. 3 liter pitcher and 20 um nitex for microplankton concentration 5. Bottles for collection of primary productivity samples (500 ml amber) 7. Van Dorn or water pump/pump tubing (only if profiles are to be taken). 8. Hydrolab (H2O) and Surveyor 3, including cable and stirrer 9. Secchi disk 10. Licor light meter, including cable 11. Field notebook (Rite-in-Rain), pencil, Sharpie 13. Raft 14. Paddles 15. Anchor bag and marked anchor line 16. Two 60 ml syringes for field filtering. 17. For each depth sampled one 500 ml amber bottle for Primary production samples. 18. Filter kit for field filtering and preservation 1. 60 ml bottle of Lugols solut | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
