Nutrient Solubilizing Microbes

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how to culture protozoa tea

  A generalized diagram on some equipment needed to cultivate protozoa

A generalized diagram on some equipment needed to cultivate protozoa

Solution A:

The "hay infusion" is perhaps the most well known culturing technique. Boil one liter of pond, spring or rain water. As the water comes to a boil, add a small handful of hay (ideally, timothy hay) and boil for ten additional minutes. The boiling will break down the hay and set up an ideal medium for the growth of bacteria. Allow this mixture to stand for two to three days. Add 25-50 milliliters (2 to 4 T.) of your sample (this is called "inoculation"). In a few days, small protozoans such as Chilomonaswill populate your culture. If Paramecium are present, they will feed on the Chilomonas and eventually increase in number (in 10 to 14 days). The organism at the top of the food chain will become the most common but will quickly die off as the food supply is exhausted. You may be successful in maintaining one organism for long periods of time by sub-culturing into newly prepared media.

Solution B:

Boil 100 milliliters of pond or spring water for ten minutes. Add five grains of wheat to the cooled water. Let this mixture stand in open dishes for one to two days, then inoculate this culture media with one or two tablespoons of your sample.

Solution C:

Follow the instructions for Solution B, but use five grains of uncooked rice instead of the wheat.

Solution D:

Hard boil an egg and grind a pinch (1/4 gram) of the yolk in a bowl with a small amount of water to form a paste. Add the paste to 1 liter of boiled pond or spring water and let stand for two days before inoculation.

Solution E:

Boil 250 ml of pond or spring water for ten minutes. Cool and add a pinch of powdered skim milk. Mix thoroughly and immediately inoculate your sample.

Solution F:

Boil 250 ml of pond or spring water for ten minutes. Cool and add 1/4 package (2 grams) of dehydrated yeast. Mix well and let stand in open containers for a few hours before inoculation. A rich culture should develop within one week.

 Different microorganisms thrive better in different culture media. If you are transferring Amoeba, use Solution C. For ciliates, try any of the above solutions. Paramecium are best cultivated in Solutions A, D and E. For Paramecium bursaria, use Solution C. For Blepharisma, try Solutions B or C.

Vorticella are cultured best in Solution D. After the solution has set for two days, pour off some of the clear top liquid and add the Vorticella. Sub-culture every two weeks by transferring Vorticella from the parent culture to a new, clear, egg yolk solution.

Stentor are best cultured with Solution C, but will also thrive in Solutions B and D.

Flagellates can be successfully cultivated in Solutions B and D. A modified Solution B can be prepared that will save time. Boil four wheat grains in 80 milliliters of pond water. Cool and add a few milliliters of sample containing the organisms that are to be cultured.

References

http://www.microscope-microscope.org/applications/pond-critters/culturing-protozoa.htm