LaMotte BOD5 User Manual

ColiQuant MF
L
Mott
Code 3-0035
Quantity Contents
2 Coliscan® MF, 20 mL bottle
20 Droppers, 3 mL, sterile
20 Dishes, with pads and lids, sterile
20 Membrane filter pads, with grid, sterile
1 Filter Apparatus with syringe, sterile
1 ColiQuant MF Manual
5 ColiQuant MF Colony Color Guide
Coliscan MF is a registered trademark of Micrology Laboratories LLC
Needed but not in cluded:
• Sterile collection container
• Sterile dilution water (if sample size is less than 10 mL)
• Sterile forceps
• Gloves
Stor age Conditions
Coliscan MF should be placed in the freezer (2 – 6°C) as soon as it is received. Coliscan MF can be refrozen if it has been thawed and not been used. Coliscan MF should be thawed at room temperature. Bottles may also be thawed for same day use by standing the bottles in warm water until the contents become liquid. Coliscan MF will keep for 6 weeks in the refrigerator or 12 months in the freezer.
Safety
• Coliscan MF is non-toxic. Additional information for all LaMotte reagents is available in the United States, Canada, Puerto Rico, and the US Virgin Islands from Chem-Tel by calling 1-800-255-3924. For other areas, call 813-248-0585 collect to contact Chem-Tel’s International access number.
• Read the entire manual before performing any tests.
• Wear safety glasses and protective gloves when appropriate.
• Wash hands thoroughly after performing tests. Avoid placing hands in contact with eyes or mouth. Do not eat, drink, or apply cosmetics during testing or clean up.
• Follow the general safety guidelines for your organization.
• Dispose of used dishes in the proper manner.
• Wipe tabletops and work areas after each use with disinfectant, such as a 1:10 dilution of household bleach.
• Testing by students: Ensure a safe field-testing experience by using the following: safety glasses for each student, clean pail or bucket for washing hands, jug of clean water for washing hands, soap (biodegradable if possible), towels, protective gloves, eye wash bottle, first aid kit.
• Keep used Petri dishes taped shut and out of the reach of small children and animals.
• Be sure that students understand the danger of treating reagents casually or endangering others through “horseplay”.
ColiQuant MF Method
Coliscan MF (membrane filter) medium is a nutrient liquid formulation which uses two color producing chemicals, one for the detection of the enzyme glucuronidase (produced by E. coli strains but not by general coliforms) and one for the detection of galactosidase (produced by all coliforms, including E. coli). A liquid sample containing E. coli and other coliforms is passed through a membrane filter to leave individual bacteria cells on the membrane filter. The filter is then placed on a pad soaked with the Coliscan MF medium. When incubated at a suitable temperature, the cells will grow into colony forming units (CFU) on the surface of the filter. If coliform (including E. coli) colonies are present, they will produce the enzyme galactosidase, which will react with its specific color producing substrate in the medium and a water insoluble pink pigment will color the colony. If E. coli colonies are present, they will also produce the enzyme glucuronidase, which will react with its specific color producing substrate in the medium and a water insoluble teal-green pigment will color the colony. However, since E. coli produces both galactosidase and glucuronidase, those colonies will be a combination of teal-green and pink pigments and will appear as some shade of blue-purple.
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Col lecting a Wa ter Sam ple
1. Remove the cap of the collection bottle. Do not touch the neck of the bottle or the
inside of the cap.
2. If you are testing stream water, face upstream while holding the bottle 5 to 6
inches below the surface.
3. When the bottle has filled, replace the cap.
4. The test can be done at the site or water samples may be collected in sterile
collection containers and transported back to the testing site. Water samples kept for more than one hour before testing should be kept on ice or refrigerated.
5. Do not reuse the sample collection bottle without sterilizing.
2
150mL
100
150mL
100
50
A - Apparatus Lid
B - Funnel
D - Filter Holder with O ring
F - Collection Container
I - Barrel
H - Plunger
J - Tubing
E - (Membrane Filter) C - (White Pad)
G - Adapter
Pro ce dure
Smaller diameter
Contains Clear
Media Film
Lid
Bottom
1. Thaw a bottle of Coliscan MF overnight at room temperature. (2 mL of Coliscan
MF is required for each sample to be tested). Bottles may also be thawed for same day use by standing the bottles in warm water until the contents become liquid.
Sample Collection and Dilution
2. Collect the water sample according to the chart below.
Environmental river, lake, pond, stream, ditch 1.0 to 5.0 mL
Drinking water well, municipal, bottled 100 mL
Generally, use 100 mL for samples suspected of having very low coliform levels such as drinking water. Use 1.0 to 5.0 mL for samples suspected of having moderate amounts of coliform, such as environmental samples (rivers, ponds, streams, lakes or ditches). Samples that are less than 10 mL will need to be diluted with 10 – 20 mL of a sterile diluent before filtering.
For optimum results, the sample should be collected within two hours before filtering. If this is not possible, the sample may be refrigerated or kept on ice for up to 24 hours.
3. Use a permanent marker or wax pencil to
label a dish with pad with the sample information.
4. Use the sterile 3 mL dropper to add 2mL of
Coliscan MF to the pad in the dish. Cover the dish with a lid.
Set up filter apparatus
5. Remove the sterile filter apparatus from the packaging.
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6. Remove the funnel and lid (A & B) from the apparatus. Use a clean forceps to
place a white, sterile pad (C) on the filter holder section (D) within the circle guides.
7. Open a sterile membrane filter envelope. Use clean forceps to remove the
membrane filter (E) from the pack. Carefully separate the membrane filter from the top and bottom protective backings. Do not tear or damage filter.
8. Place the membrane filter, grid side up, on top of the white pad.
9. Replace the funnel and lid on the filter holder section. Push the funnel down onto
the filter holder section until it is fully and evenly seated. This will seal the connection and hold the membrane filter and white pad firmly in place.
10. Push the plunger (H) all the way into the barrel (I) of the syringe. Then attach the
tubing (J) from the syringe to the adapter (G) on the side of the collection container (F).
Filter the sample
11. Swirl the water sample container to mix the contents.Remove the apparatus lid
from the funnel. Pour the water sample into the funnel.
If the sample size is less than 10 mL, add 10-20 mL of sterile diluent to the funnel before attaching the syringe. Then add the sample and swirl the apparatus gently to mix the sample and the diluent. Attach the syringe.
NOTE: The diluent is used to distribute the sample more evenly over the surface of the membrane filter during filtration. The amount of diluent should not be included in the final calculation. Use only the amount of the original sample size in the calculation.
12. Pull the syringe plunger out, nearly to the end of the barrel, to create a vacuum.
The water sample will be pulled through the membrane filter. Any microorganisms that are present will be deposited on the surface of the membrane filter.
NOTE: Larger water samples may not be completely filtered with a single pull of the plunger. To complete the filtration, carefully remove the tubing and syringe from the adapter on the side of the filter apparatus. Push the plunger all the way into the barrel of the syringe. Reattach the tube to the apparatus and pull out the plunger.
13. When the entire water sample has passed through the membrane filter, remove
the tubing and syringe. Carefully remove the funnel from the apparatus.
14. Use clean forceps to lift the membrane filter from the filter holder section of the
apparatus. Place the membrane filter, grid side up, on top of the pad in a
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