Tokophrya lemnarum (Stein) Entz


Product category
KINGDOM: Protozoa
Strain designation
Type strain
Isolation source
Wastewater treatment plant
Geographical isolation
United States; New York; Oneonta
Product format
Test tube
Mission Collection Item
This is a Mission Collection Item


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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed product information



Handling information

Instruction for complete medium
ATCC Medium 1323
ATCCNO: 50032 SPEC: Food source, Paramecium tetraurelia ATCC 30567, not supplied.
Handling notes
This strain must be fed with live Paramecium (i.e., ATCCÒ 30567™ or similar, not provided).  The Paramecium should be maintained separately and fed to Tokophrya at regular intervals.  Overfeeding of Tokophrya may result in monster formation.  Attempt to maintain a ratio of 2-3 prey organisms per each suctorian.  If the number of abnormal suctorians is high, reduce the feeding interval or passage the culture.

This strain of Tokophrya lemnarum is mating type I.  The culture is polyxenic and contains mixed bacterial flora.

Culture maintenance
Periodically add prey organisms as follows:

1.   Maintain growing cultures of Paramecium separately at 25°C in T-25 tissue culture flasks containing 10 ml ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC 13048).

2.   Prepare washed Paramecium as follows:  Remove 5-10 ml from a culture at or near peak density, centrifuge at 300 x g for 5 min, quickly remove most of the supernatant (leaving approx. 1 ml), then resuspend cells in 10 ml ATCC medium 1323.  Centrifuge and resuspend cells again as above.  Repeat this washing step at least twice.

3.   When the Tokophrya have consumed all prey Paramecium, add 0.5-2 ml of washed Paramecium prepared in step 2.  The feeding interval will depend on the number of suctorians present and the culture density of the washed prey.

4.   The Tokophrya may be passaged to a new petri plate or T-25 flask by gently rubbing the agar surface with a spread bar to dislodge attached suctorians, then transferring 0.5 to 2 ml to a fresh petri plate or T-25 flask containing a bed of non-nutrient agar (ATCC medium 919) and 10 ml ATCC medium 1323.  Incubate the culture at 20-25°C, feeding periodically with washed Paramecium.

Reagents for cryopreservation
Cryoprotective Solution

DMSO                                                                                    2.0 ml

Fresh growth medium w/o bacteria                                 8.0 ml

1.     Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.

2.    Harvest Tokophrya cells from a culture that has recently passed peak density by centrifugation at 250-300 x g for 5 min.

3.     Adjust the concentration of cells to at least 2 x 104/ml in fresh medium.

4.     Mix the cell preparation and the cryoprotective solution in equal portions by adding the cryoprotective solution to the cell suspension in 3 equal aliquots at 2 min. intervals.

5.     Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

6.     Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1 C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  

7.     Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8.     To establish a culture from the frozen state place the vial in a 35°C water bath. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial. Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and transfer to a petri plate or T-25 tissue culture flask containing a bed of non-nutrient agar (ATCC medium 919) and 10 ml ATCC medium 1323.

9.     Aseptically transfer 0.5-2.0 ml of washed Paramecium to the petri plate or T-25 flask (see section on MAINTENANCE OF CULTURE).  Incubate the culture at 20-25°C.

Once the culture is established, follow the protocol for maintenance of culture.


Deposited as
Tokophrya lemnarum (Stein) Entz
LA Colgin-Bukovsan
Year of origin

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The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.


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