This video documents large volume (≥20 L) filtration of microbial biomass, ranging between 0.22μm and 2.7μm in diameter, from the water column.
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Method Article
Large Volume (20L+) Filtration of Coastal Seawater Samples
In This Article
Summary
Abstract
The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For this method, samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents large volume (≥20 L) filtration of microbial biomass, ranging between 0.22μm and 2.7μm in diameter, from the water column. Two 20L samples can be filtered simultaneously using a single pump unit equipped with four rotating heads. Filtration is done in the field on extended trips, or immediately upon return for day trips. It is important to record the amount of water passing through each sterivex filter unit. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use. This procedure will take approximately 5 hours plus an additional hour for clean up.
Protocol
- Set up of filtration apparatus
- To set up the filtration apparatus, use tweezers to place a grade GF/D filter into the prefilter housing unit. Loosely tighten the housing by hand. As a rule of thumb, do not over-tighten or the unit will leak once the pumping starts.
- Replace the carboy cap with the autoclaved cap and attached tubing by carefully inserting the internal tubing into the carboy without touching it. If necessary, use tweezers to guide the tubing into the carboy.
- Pass the external tubing through the peristaltic pump.
- Attach prefilter housing units to the end of both external tubes (i.e. 2 units per carboy....
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Discussion
It is imperative to wipe down benches and pumps with a damp towel after use to remove any lingering traces of salt water that would otherwise corrode equipment. Although the procedure is simple, the time requirement can be extensive depending on the concentration of biomass or debris in the sample being filtered.
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Disclosures
The authors have nothing to disclose.
Acknowledgements
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. D.A.W. was supported by fellowships from NSERC, Killam and the TULA foundation funded Centre for Microbial Diversity and Evolution.
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Materials
| Name | Company | Catalog Number | Comments | |
|---|---|---|---|---|
| Peristaltic pump | Masterflex (Cole Palmer) | 77913-20, 77200-62, 77200-04 | ||
| Tygon LFL tubing | Masterflex (Cole Palmer) | 06429-24 | For filtration apparatus | |
| Hose clamps | Cole-Parmer | 0683204 | For filtration apparatus | |
| 1/4” x 3/16” pipe adaptors | Cole-Parmer | 31702-20 | For filtration apparatus | |
| Polypropylene prefilter housing | ADVANTEC | 501200 | For filtration apparatus | |
| Metal luer fitting (Hose end to Male luer lock) | Cole-Parmer | 31507-27 | For filtration apparatus | |
| 2“ Magnets | For filtration apparatus | |||
| 3 port carboy caps | For filtration apparatus | |||
| GV 0.22 μm STERIVIX filters | EMD Millipore | SVGV01R5 | ||
| 47 mm diameter 2.7 μm GF/D filters | Whatman, GE Healthcare | 1823047 | ||
| Lysis buffer | Reagent | 0.75 M sucrose, 40 mM EDTA, 50 mM Tris (pH 8.3) | ||
| 0.1 % HCl | Reagent | For post-filtration rinse | ||
| Autoclaved water | Reagent | For post-filtration rinse |
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