Tokophrya lemnarum (Stein) Entz
50033 ™
- Product category
-
Protists
- Classification
- KINGDOM: Protozoa
- Strain designation
- Oneonta
- Type strain
- No
- Isolation source
- Wastewater treatment plant
- Geographical isolation
- United States; New York; Oneonta
- Product format
- Test tube
Documentation
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
Characteristics
- Comments
-
II
Handling information
- Medium
- Instruction for complete medium
- ATCC Medium 1323
- Temperature
-
25°C
- Incubation
- 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
- Cryopreservation
- 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.
History
- Deposited as
- Tokophrya lemnarum (Stein) Entz
- Depositors
- LA Colgin-Bukovsan
- Year of origin
- 1982
Legal disclaimers
- Intended use
- This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
- Warranty
-
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.
- Disclaimers
-
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
Permits & Restrictions
Import Permit for the State of Hawaii
If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.
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