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Acanthamoeba pearcei Nerad et al.

50435

Product category
Protists
Classification
Amoebozoa, Acanthamoebidae
Strain designation
205-1
Type strain
Yes
Isolation source
Seabottom sample from depth of 50 m at site used for disposal of barge-delivered wastes from 1973-1980 in Atlantic Ocean approximately 65 km seaward from the coast of Delaware and Maryland (38 degrees 19.6'N, 74 degrees 18.0'W)
Geographical isolation
United States
Product format
Dried
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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

Medium
Instruction for complete medium

Media: ATCC medium 997 optionally inoculated with Klebsiella pneumoniae subsp. pneumoniae (ATCC 700831™) or Enterobacter aerogenes (ATCC 13048ä).

Alternate media: ATCC medium 711 optionally inoculated with bacteria as above.  (Note that ATCC medium 711 is a richer formula than ATCC medium 997 and will produce denser and faster bacterial growth.  Excess bacterial growth can inhibit growth of amoebae.)

Temperature
25°C
Culture system
Xenic
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 culture will not be rehydrated within that period, store at 5°C until processed.

  1. To rehydrate an ampule, aseptically add 1.0 ml of sterile distilled water to the inner shell vial.  Aseptically remove the filter paper pellet with a pair of sterilized forceps and place it in the center of a plate of ATCC medium 997.
  2. Using a Pasteur pipette, aseptically transfer the remainder of the liquid from the vial to the plate.  Tease apart the filter paper pellet, and distribute evenly over the surface of the plate using a spread bar.
  3. Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.  Trophozoites should be seen within 5-7 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.

Reagents for cryopreservation

Cryoprotective Solution
DMSO, 1.5 ml
Dryl's solution (or similar), 8.5 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 cells from a culture which is at or near peak density by adding 5 ml ATCC medium 5080 (Dryl's solution) and washing cells into suspension.  Rub the surface of the plate with a spread bar to detach adhering trophozoites.
  3. Adjust the concentration of cells to at least 2 x 106/ml in fresh medium.
  4. Mix the cell preparation and the cryoprotective solution in equal portions.
  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, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.  Distribute the material evenly over the plate using a spread bar.
  9. Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.

Follow the protocol for maintenance of culture.

History

Deposited as
Acanthamoeba pearcei Nerad et al.
Depositors
TK Sawyer
Type of isolate
Environmental
Year of origin
1973
Cross references
GenBank AF019053 Acanthamoeba pearcei 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.
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.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Nerad TA, et al. Acanthamoeba pearcei n. sp. (Protozoa: Amoebida) from sewage contaminated sediments. J. Eukaryot. Microbiol. 42: 702-705, 1995. PubMed: 8520585

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

type strain

Gast RJ. Development of an Acanthamoeba-specific reverse dot-blot and the discovery of a new ribotype. J. Eukaryot. Microbiol. 48: 609-615, 2001. PubMed: 11831768

Marciano-Cabral F, Cabral G. Acanthamoeba spp. as agents of disease in humans. Clin. Microbiol. Rev. 16: 273-307, 2003. PubMed: 12692099

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