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Acanthamoeba stevensoni Sawyer et al.


Product category
Amoebozoa, Acanthamoebidae
Strain designation
Type strain
Isolation source
Hard clam beds, Mercenaria mercenaria, contaminated by sewage; Raritan Bay, near the Staten Island shoreline
Geographical isolation
United States; New York
Product format
Storage conditions
2°C to 8°C
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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


species description

Handling information

Instruction for complete medium
ATCC Medium 711
Culture system
Handling procedure

Establishing Cultures from Dried State

This strain comes dried on shredded filter paper.  Dried samples can remain at room temperature for up to one week.  If the cultures will not be rehydrated within that period, store at 5°C until processed.

  1. To rehydrate an ampule, aseptically add 1 ml of sterile distilled water to the inner shell vial.  Aseptically remove the filter paper pellet with a pair of forceps, and place it in the center of a plate of ATCC medium 711.
  2. Add the liquid remaining in the vial to the plate, tease apart the filter paper pellet, and distribute evenly over the surface of the plate.
  3. Incubate the plate upright at 25°C.  Trophozoites should be seen within 2-3 d.

Culture maintenance
  1. Streak an ATCC medium 711 plate with Enterobacter aerogenes (ATCC® 13048) and incubate at 35°C overnight.
  2. Remove an agar block (~5 mm2), with trophozoites or cysts, from the edge of an agar plate culture and invert the block at the edge of the freshly bacterized plate.
  3. Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.
  4. Repeat steps 1-3 at 10-14 d intervals.

Note: a monoxenic amoeba culture can be established in this manner using any suitable bacterial food source.

  1. Allow the cells to encyst.  To detach cysts from the plate flush the surface with 5 ml fresh ATCC medium 1323 (Page's Balanced Salt Solution).  Rub the surface of the plate with a spread bar to detach adhering cysts.
  2. Transfer the liquid medium to a sterile centrifuge tube.
  3. If the cyst concentration does not exceed 2 x 106 cysts/ml adjust the suspension to that concentration.  To adjust the concentration, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield 2 x 106.
  4. While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO as follows:  Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath.  Allow the DMSO to solidify.  Add the required volume of refrigerated medium.  Dissolve the DMSO by inverting the tube several times. *NOTE: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
  5. Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be at least 106 cysts/ml and 7.5% (v/v) DMSO.  The equilibration time (the time between addition of DMSO and the start of the cooling cycle) should be no less than 15 min and no longer than 30 min.
  6. Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
  7. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  If the freezing unit can compensate for the heat of fusion, maintain rate at        -1°C/min through the heat of fusion.  At -40°C plunge 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.)  
  8. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.
  9. To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min). Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.
  10. Immediately after thawing, aseptically remove the contents of the ampule and distribute to the center of a fresh plate of ATCC medium 711.  Distribute the material evenly over the plate using a spread bar.  Incubate at 25°C.


Deposited as
Acanthamoeba stevensoni Sawyer et al.
TK Sawyer
Type of isolate
Animal; Wastewater
Special collection
NSF - Protistology
Cross references
GenBank AF019069 Acanthamoeba stevensoni RB:F:1 18S ribosomal RNA gene, partial sequence.

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.

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.


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

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.


Frequently Asked Questions


Curated Citations

Sawyer TK, et al. Acanthamoeba stevensoni n. sp. (protozoa: amoebida) from sewage-contaminated shellfish beds in Raritan Bay, New York. J. Eukaryot. Microbiol. 40: 742-746, 1993.

Stothard DR, et al. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J. Eukaryot. Microbiol. 45: 45-54, 1998. PubMed: 9495032

Kong HH, et al. Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small-subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. J. Clin. Microbiol. 40: 1199-1206, 2002. PubMed: 11923331

type strain

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