Name: Seawater Sampling and Collection (Video) | JoVE Journal
Uploaded: 2009-06-17T19:00:00.000+00:00
Duration: 8 min 23 s
Description: Learn seawater sampling methods for marine biology and environmental science using go flow and kin water sampling devices. This procedure enables researchers to collect samples for biomass, nucleic acid, and nutrient analyses.

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5 minutes

We would like to thank the Canadian Foundation for Innovation, the British Columbia Knowledge Development Fund and the National Sciences and Engineering Research Council (NSERC) of Canada for supporting ongoing studies on low oxygen regions of coastal and open ocean waters. In addition, D.A.W. was supported by fellowships from NSERC, Killam and the TULA foundation funded Centre for Microbial Diversity and Evolution.

Part 1: Water Collection
<ol>
<li>Depths are assigned to each cast.</li>
<li>Depths to be sampled and the total volume to be collected for the current cast are determined.</li>
<li>It is ensured that the Go-Flo and Niskin bottles are open, and that their valves and spigots are closed.</li>
<li>The CTD is loaded onto bottom of the line and lower into water.</li>
<li>A bottle for the deepest depth sampled in cast first is loaded onto the line after the CTD; it is ensured that the line is properly fitted into bracket and the screws are tightened.</li>
<li>A messenger is attached to each bottle (except the deepest one) before lowering.</li>
<li>The line is lowered the distance between the first depth and the next; another bottle is then loaded. It should be noted that for depths where 20L are collected for filtration, a total of 2 12L bottles are loaded consecutively about .75m apart.</li>
<li>Once all the bottles are on the line, the line is lowered to the desired depth and 2 minutes are allowed to pass.</li>
<li>A messenger is sent down and a few minutes are allowed to pass to ensure that all bottles close.</li>
<li>The line is reeled in and the bottles are unloaded as they come up onto the racks.</li>
<li>For depths that require 20L samples for DNA and RNA, two Niskin bottles are collected. One bottle is used to sample the chemical parameters (see below), while the other is drained into the carboy. RNA is collected immediately from the second Niskin. The remainder of the first is also drained into the carboy to give ~20L final volume. For low-resolution samples only 1L of water is set aside for filtration in sterile Nalgene bottles.</li>
</ol>
Part 2: Gas sample collection
<ol>
<li>If oxygen samples are needed, they are collected first.</li>
<li>A piece of thin tubing is attached to the Niskin/Go-Flo spigot and the other end of the tubing is placed place into the glass serum bottles.</li>
<li>The serum bottle is rinsed 3 times with sample water, and filled to overflowing such that no air bubbles are present.</li>
<li>50&mu;l mercury chloride (HgCl2) are added to the serum bottle in a way that doesn't introduce air bubbles into the sample.</li>
<li>The bottle is sealed with a rubber stopper and crimped with an aluminum cap, and is then stored on ice in a cooler.</li>
</ol>
Part 3: Nutrient sample collection
<ol>
<li>A 60 cc syringe is rinsed with sample water, an (0.2 &mu;m) is attached in-line and some water is pushed through the acrodisc.</li>
<li>A polyethylene bottle is rinsed with ~10 ml filtered water 3 times (shake bottle when rinsing).</li>
<li>Filtered sample water is added to the bottles, about &frac34; full.</li>
<li>The sample is stored on ice in the cooler.</li>
</ol>
Part 4: Ammonia sample collection
<ol>
<li>Sample tubes are rinsed with sample water.</li>
<li>Sample tubes are filled completely with sample water.</li>
<li>5ml of sample water are taken out using a 10ml pipet; the rest of the water in the tube is discarded and the 5ml in the pipet tip are replaced into the sample tube.</li>
<li>7.5ml of ammonium working reagent is added to each tube once samples from all depths have been collected.</li>
</ol>
Part 5: Sample collection for cell enumeration
<ol><li>A pre-labelled 50 ml Falcon tube containing 5.4 ml 37%formaldehyde is filled to the top for a final concentration of 4%. Care is taken not to spill.</li></ol>
Part 6: Sample collection for hydrogen sulphide measurements
<ol>
<li>A pre-labelled 50 ml Falcon tube containing 20% zinc acetate (Zn(O2CCH3)2) is filled to the top. Care is taken not to spill.</li>
</ol>
Part 7: Small volume collection for biomass concentration
<ol>
<li>1-litre bottles are filled to the top.</li>
</ol>
Part 8: Large volume collection for biomass concentration
<ol>
<li>20-litre carboys are filled.</li>
</ol>
Part 9: Representative Results
The final outcome of seawater sampling results in approximately 12 dissolved oxygen samples, 32-48 gas samples, 16 nutrient samples, 32 ammonium samples, 16 hydrogen sulfide samples, 16 samples for cell counts, 6 20-litre carboys, 16 1-litre bottles and 4 Sterivex 0.22&mu;m filters containing microbial biomass preserved with RNA later.

<ol><li>Cline, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spectrophotometric+determination+of+hydrogen+sulfide+in+natural+waters%5BTitle%5D)+AND+%22Limnology+%26+Oceanography%22%5BJournal%5D)">Spectrophotometric determination of hydrogen sulfide in natural waters.</a> Limnology & Oceanography. 14, 454-458 (1969).</li>
<li>Holmes, R. M., Aminot, A., Kerouel, R., Hooker, B. A., Peterson, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+simple+and+precise+method+for+measuring+ammonium+in+marine+and+freshwater+ecosystems%5BTitle%5D)+AND+%22Canadian+Journal+of+fisheries+and+Aquatic+Science%22%5BJournal%5D)">A simple and precise method for measuring ammonium in marine and freshwater ecosystems.</a> Canadian Journal of fisheries and Aquatic Science. 56, 1801-1808 (1999).</li>
</ol>

The authors have nothing to disclose.

Overall, there is a great deal of preparation and planning required for this type of sampling, so budget 2 days before the trip to get things in order and one day afterwards for cleaning everything up. In general, do not let salt water come into contact with the peristaltic pump or into the aluminum Zarges box used to transport the field equipment. Treat the equipment with respect and it will perform well for a long time to come.

<table><tbody><tr><td>20L Carboys</td><td></td><td>Fisher Scientific</td><td>02-960-15</td><td></td></tr><tr><td>3/8&quot; Tygon</td><td></td><td>Cole-Parmer</td><td>06429-36</td><td></td></tr><tr><td>Sterivex filters</td><td></td><td>Fisher Scientific</td><td>SVG010RS</td><td></td></tr><tr><td>Prefilter Housing Unit</td><td></td><td>AMD</td><td>501200</td><td></td></tr><tr><td>Luer Fitting</td><td></td><td>Cole-Parmer</td><td>31507-27</td><td></td></tr><tr><td>1/4&quot; Tygon tubing</td><td></td><td>Cole-Parmer</td><td>06429-24</td><td></td></tr><tr><td>1/4&quot; Tygon tubing</td><td></td><td>Cole-Parmer</td><td>06429-24</td><td></td></tr><tr><td>1/4&quot; Tygon tubing</td><td></td><td>Cole-Parmer</td><td>06429-24</td><td></td></tr><tr><td>50ml Plastic tubes</td><td></td><td></td><td></td><td></td></tr><tr><td>Glass Serum Bottles</td><td></td><td></td><td></td><td>Glass Sample Collection</td></tr><tr><td>Septa and aluminum caps for glass serum bottles</td><td></td><td></td><td></td><td></td></tr><tr><td>Hand Crimper</td><td></td><td></td><td></td><td></td></tr><tr><td>20-200ul Pipette and Tips</td><td></td><td></td><td></td><td></td></tr><tr><td>Glass Tubes</td><td></td><td></td><td></td><td>Ammonium Sample Collection</td></tr><tr><td>30ml Plastic Bottles</td><td></td><td></td><td></td><td>Nutrient Collection</td></tr><tr><td>Acrodisc Filters</td><td></td><td>EMD Millipore</td><td></td><td></td></tr><tr><td>5 ml Tubes</td><td></td><td></td><td></td><td>Nitrite Sample Collection</td></tr><tr><td>37% Formaldehyde</td><td>Reagent</td><td></td><td></td><td>For cell count samples</td></tr><tr><td>20% Zinc Acetate</td><td>Reagent</td><td></td><td></td><td>For sulfide samples</td></tr><tr><td>Mercuric Chloride</td><td>Reagent</td><td></td><td></td><td>For gas samples</td></tr><tr><td>milliQ water</td><td>Reagent</td><td></td><td></td><td></td></tr><tr><td>Ammonium Working Reagent</td><td>Reagent</td><td></td><td></td><td>Borate buffer, sodium sulfite, orthophaladialdehyde</td></tr><tr><td>Sulfanilamide</td><td>Reagent</td><td></td><td></td><td>5g into 450ml water and 50ml HCl</td></tr><tr><td>N-(1-naphthy)-ethylenediamine dihydrochloride solution</td><td>Reagent</td><td></td><td></td><td>0.5g into 500ml water</td></tr></tbody></table>

seawater sampling and collection

Seawater Sampling and Collection

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass concentration, nucleic acid purification, cell abundance, nutrient and trace gas analyses. For today's demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. An A-frame derrick, with a multi-purpose winch and cable system, is used in combination with Niskin or Go-Flo water sampling bottles. Conductivity, Temperature, and Depth (CTD) sensors are also used to sample the underlying water mass. To minimize outgassing, trace gas samples are collected first.  Then, nutrients, water chemistry, and cell counts are determined. Finally, waters are collected for biomass filtration. The set-up and collection time for a single cast is ~1.5 hours at a maximum depth of 215 meters. Therefore, a total of 6 hours is generally needed to complete the collection series described here.

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass concentration, nucleic acid purification, cell abundance, nutrient and trace gas analyses.

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass ...

Hi, I'm Elena Kova from the Laboratory of Stephen Hallam in the Department of Microbiology and Immunology at the University of British Columbia. Today we'll show you a procedure for collecting sea water for physical, chemical and biological analysis. We use this procedure in our laboratory to study ocean graphic parameters of marine waters.So let's get started. First, the number of casts and depths in each cast are assigned, then depths to be sampled, and the total volume to be collected for the current cast are determined. Ensure that the go flow and kin water sampling devices are in the open position and that all valves and spigots are closed.Attach the CTD to the base of the line and lower it into the water for the deepest depth in the profile. Bring a sampling device onto the line immediately following the CTD. Make sure that the line is properly fitted into the device bracket and the screws are tightened.Attach a weighted messenger to each device except for the deepest sampling depth before lowering it into the water. Lower the line and the distance between the first specified depth and the next, then rig another sampling device. Note that for depth intervals where large volume water samples are collected, a total of two x 12 liter bottles are loaded consecutively approximately 0.75 meters apart.After all the devices are on the line, lower the line to the deepest depth and allow two minutes to pass. Send a messenger down, tripping each device into the close position. Allow a few minutes to pass before recovering the line.To ensure that all devices are closed, reel the line in and unload each device onto the deck as it emerges from the water. For depths that require large sample volumes for protein DNA or RNA purification, employ two sampling devices in tandem. Collect RNA immediately from one bottle and filter it on ship.After sampling chemicals and nutrients in the other bottle, drain the remaining water into the car to yield a 20 liter final volume. For low volume samples, set aside one liter of water for filtration in sterile Nalgene bottles. Typically low volume samples from 16 depth intervals are collected as 16 x one liter samples and six x 20 liter samples are collected as high volume samples.When collecting gas samples, always take oxygen and trace gas samples. First, attach a thin piece of tubing to the spigot of the water sampling device. Place the other end of the tubing into the glos serum bottles.Rinse the serum bottle three times, but sample water and fill it to overflowing such that no air bubbles are present. Add 50 microliter mercuric chloride to the serum bottle. Be careful not to introduce air bubbles into the sample.Seal the serum bottle with a rubber stopper C crimp with an aluminum cap and then store it on ice to collect samples for nutrient measurements including nitrate, phosphate and silicate. First rinse a 60 cc syringe with sample water. Then attach a 0.2 micrometer acro disc filter in line.Push approximately two ml water through the syringe. To purge the filter. Rinse a polyethylene bottle with approximately 10 ml of filtered water three times.Shake the bottle when rinsing, then add filtered sample water to the bottles about three quarters full. Store the sample on ice. Empty collection tubes of distal water to collect samples for ammonia measurements.First rinse collection tubes, but sample water completely. Fill a 10 ml graduated cylinder with sample water pipe at five ml of sample water out of the graduated cylinder and empty the remainder from the cylinder. Then add the five ml back to the empty cylinder.Once samples from all depths have been collected at 7.5 ml of ammonium working reagent to each tube. To collect samples for cell counts, fill a pre-labeled 50 ML Falcon tube containing 5.4 ml, 37%formality height to the 50 ml mark for a final concentration of 4%Take care not to overflow the tube. Store the sample on ice to collect samples.For hydrogen sulfide measurements. Fill a pre-labeled 50 ML Falcon tube containing three ml, 20%zinc acetate to the top. Take care not to overflow the tube.Store the sample on ice to collect small volumes. For biomass concentration. Fill one liter Nalgene bottles to the top to collect large volumes for biomass concentration.Fill 20 liter carboys to the top. The final outcome of seawater sampling results in approximately 32 to 48. Gas samples, 32 ammonium samples.16 hydrogen sulfide samples. 16 samples for cell counts. Six 20 liter car, 16 one liter bottles, and four sterex.Point two two micrometer filters containing microbial biomass are preserved for later. RNA isolation. We've just demonstrated how to collect different kinds of water samples for biochemical analysis.This requires a great deal of preparation and planning. So budget two days before the trip to get things in order and one day afterwards for cleanup, do not let salt water come into contact with a peralt pump or into the aluminum zago box used to transport the field equipment. Treat the equipment with respect and it will serve well for a long time to come.So that's it. Thanks for watching and good luck with your experiments.

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Learn seawater sampling methods for marine biology and environmental science using go flow and kin water sampling devices. This procedure enables researchers to collect samples for biomass, nucleic acid, and nutrient analyses.

Seawater Sampling and Collection

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