Tetrahymena malaccensis Simon et al.


Product category
KINGDOM: Protozoa
Strain designation
Type strain
Isolation source
Jungle stream near campus of University Pertanian Malaysia
Geographical isolation
Malaysia; Selangor; Serdang
Product format
Test tube
Storage conditions
See handling procedure
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.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Modified PYNFH Medium [ATCC medium 1034]


Price: $74.00 ea

Detailed product information


Specific applications
Identification of Tetrahymena species using PCR/RFLP analysis of rDNA


Identification of Tetrahymena species using PCR/RFLP analysis of rDNA
Species description
Ribosomal RNA introns

Handling information

Instruction for complete medium
Media: ATCC Medium 1034 without serum is used for short-term cultivation. (ATCC Medium 1034 is available in a freeze-dried format as cat. no. 327-X).

Alternate Media: ATCC Medium 357 can also be used for short-te
Culture system
Handling procedure
Handling of Live Culture
This strain is routinely shipped as a growing culture in a glass 16 x 125 mm screw-capped test tube.  The volume of the cell suspension is approximately 5 mL.  When the culture arrives remove it promptly from the shipping container.  Do not store the culture at refrigeration temperatures before handling.  To assure viability, immediately loosen the test tube cap and incubate upright at 25°C for at least one hour before observing the culture.  There should be numerous active trophozoites in suspension.  If the numbers are low the culture may have been exposed to temperature extremes in transit.  Regardless of the state of the culture, aseptically transfer a 0.5 mL aliquot to a 16 x 125 mm screw-capped test tube containing 5 mL of sterile ATCC medium 1034 or 357.  Incubate the parent and daughter cultures upright with the caps on loosely at 25°C.
Handling notes
Serum should be omitted from ATCC medium 1034 for cultivation of Tetrahymena spp.  The serum will not harm the culture over the short term, but it is richer in nutrients than the Tetrahymena require, and they may become more dependent upon it over the long term.
Culture maintenance

Routine Short-term Maintenance:

  1. Inoculate 5.0 mL of ATCC medium 1034 or 357 with 0.1 mL from a Tetrahymena culture at or near peak density.
  2. Incubate upright at 25°C with cap loosened one half turn.
  3. For routine maintenance subculture every 10-14 d.


Routine Long-term Maintenance:

  1. Screw the cap on tightly of a Tetrahymena culture at or near peak density and invert several times.  Aseptically transfer 0.1 mL aliquot to a fresh vessel of ATCC medium 383.
  2. Loosen caps one half turn and incubate vertically at 25°C for 1 d.
  3. Place cultures at 15°C for 3-6 wk.
  4. For routine maintenance subculture every 3-6 wk.
Reagents for cryopreservation
RM-9 Media for cryopreservation of Tetrahymena
Proteose Peptone (Difco 0120) 5.0 g
Tryptone 5.0 g
K2HPO4 0.2 g
Glucose 1.0 g
Liver extract 0.1 g
Glass distilled water 1.0 L

Dissolve components in glass distilled H2O and autoclave.

Dryl’s Salt Solution
0.1 M NaH2PO4 . 3H20 10.0 mL
0.1 M Na2HPO4 . 7H20 10.0 mL
0.1 M Sodium citrate . 2H20 15.0 mL
0.1 M CaCl2 . 2H20 15.0 mL
Distilled water 950.0 mL
Add the first 3 components to the distilled H2O and mix thoroughly. Add the CaCl2 solution and mix thoroughly. (Adding the solutions in the order indicated will avoid the precipitation of Ca salts.)

Harvest and Preservation

  1. Transfer Tetrahymena from usual growth medium to RM-9 medium and allow to grow to near peak density.
  2. Harvest cells from a culture by centrifugation at 300 x g for 2 min.          
  3. Adjust concentration of cells to 2 x 106/mL in fresh medium.
  4. While cells are centrifuging, prepare a 22% (v/v) sterile solution of sterile DMSO in fresh medium.
    1. Add 2.2 mL of DMSO to an ice cold 20 x 150 mm screw-capped test tube;
    2. Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 7.8 mL of ice cold medium;
    3. Invert several times to dissolve the DMSO;
    4. Allow to warm to room temperature.
  1. Add a volume of the DMSO solution equal to the cell suspension volume but add in 3 equal aliquots at 2 min intervals.  Thus, the final concentration of the preparation will equal 11% (v/v) DMSO and 106 cells /mL.
  2. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
  3. Place the ampules in a controlled rate freezing unit.  The cooling cycle should be initiated no less than 15 min and no longer than 60 min after the addition of the DMSO to the cell preparation.  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 -50°C ampules are plunged into liquid nitrogen.
  4. Store in the vapor or liquid phase of a nitrogen refrigerator.
  5. To establish a culture from the frozen state aseptically add 0.5 mL sterile Dryl's Salt Solution to an ampule.  Immediately place the ampule in a 35°C water bath until thawed (2-3 min).  Immerse the ampule just sufficiently to cover the frozen material.  Do not agitate the ampule.
  6. Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5.0 mL of fresh medium in a 16 x 125 mm screw-capped test tube with a slightly loosened cap.  Incubate at 25°C.


Alternative Thawing Procedure

  1. Aseptically add 0.5 mL of sterile modified PYNFH medium (ATCC Medium 1034) containing 8% (w/v) sucrose to the ampule.  Immediately, place in a 35°C water bath, until thawed.  Immerse the ampule just sufficient to cover the frozen material.  Do not agitate the ampule.
  2. Immediately after thawing, aseptically remove the contents of the ampule and gently add the material to the edge of a 20 x 100 mm petri plate containing ATCC Medium 919 (non-nutrient agar) and position on a 15 degree slant.  The cell suspension will pool at the edge of the plate.
  3. Continue to double the volume of the cell suspension at 10 minute intervals by adding ATCC medium 1034 containing 4% sucrose (w/v).  When the volume reaches 16.0 mL place the plate in horizontal position and incubate at 25°C. 
  4. On the following day, gently remove the cell suspension for the plate and transfer to a T-25 tissue culture flask.  Note the volume of the suspension and add a volume of fresh medium containing 4% sucrose equal to the volume of the cell suspension.  Incubate the culture at 25°C.
  5. After the culture has been established subculture into fresh medium without sucrose.


Deposited as
Tetrahymena malaccensis Simon et al.
EM Simon, E Meyer
Type of isolate
Year of origin

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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.

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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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.



Curated Citations

Simon EM, et al. New wild Tetrahymena from Southeast Asia, China, and North America, including T. malaccensis, T. asiatica, T. nanneyi, T. caudata, and T. silvana n. spp.. J. Protozool. 32: 183-189, 1985. PubMed: 3989748

Jerome CA, Lynn DH. Identifying and distinguishing sibling species in the Tetrahymena pyriformis complex (Ciliophora, Oligohymenophora) using PCR/RFLP analysis of nuclear ribosomal DNA. J. Eukaryot. Microbiol. 43: 492-497, 1996. PubMed: 8976607

Sogin ML, et al. Phylogenetic evidence for the acquisition of ribosomal RNA introns subsequent to the divergence of some of the major Tetrahymena groups. EMBO J. 5: 3625-3630, 1986. PubMed: 3830129

Nanney DL, et al. Comparison of sequence differences in a variable 23S rRNA domain among sets of cryptic species of ciliated protozoa. J. Eukaryot. Microbiol. 45: 91-100, 1998. PubMed: 9495037

type strain

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