Acanthamoeba castellanii (Douglas) Page (ATCC® 30011)

Depositor: A Castellani  /  Biosafety Level: 1

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Biochemical and molecular characterization
characterization of Acanthamoeba polyphaga
Molecular characterization of corneal pathogen
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Isolation Yeast culture, London, 1930
Product Format frozen
Storage Conditions Frozen Cultures:
-70°C for 1 week; liquid N2 vapor for long term storage

Freeze-dried Cultures:

Live Cultures:
See Protocols section for handling information
Type Strain yes
Two strains from human tissue
Reduction of uptake of Legionella by antimicrobial agents
Two genetic markers that distinguish pathogenic and nonpathogenic strains
Restriction fragment length polymorphisms of DNA
Pathogenicity and isoenzyme profiles
Biochemical and molecular characterization
Mitochondrial DNA fingerprinting
Adherence characteristics
Characterization of Acanthamoeba polyphaga
Temperature-dependent replication of Legionella pneumophila
Molecular characterization of corneal pathogen
Medium ATCC® Medium 997: Fresh water ameba medium
ATCC® Medium 711: PYB
Growth Conditions
Temperature: 25°C
Culture System: Xenic
Cryopreservation Reagents
Cryoprotective Solution
DMSO, 1.5 mL
Dryl's solution (or similar), 8.5 mL

Harvest and Preservation
  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. 
Name of Depositor A Castellani
Year of Origin 1930

Page FC. Re-definition of the genus Acanthamoeba with descriptions of three species. J. Protozool. 14: 709-724, 1967. PubMed: 5604481

Moura H, et al. Acanthamoeba healyi n. sp. and the isoenzyme and immunoblot profiles of Acanthamoeba spp., groups 1 and 3. J. Protozool. 39: 573-583, 1992. PubMed: 1522539

Visvesvara GS, et al. Isolation of two strains of Acanthamoeba castellanii from human tissue and their pathogenicity and isoenzyme profiles. J. Clin. Microbiol. 18: 1405-1412, 1983. PubMed: 6655045

McLaughlin GL, et al. Restriction fragment length polymorphisms of the DNA of selected Naegleria and Acanthamoeba amebae. J. Clin. Microbiol. 26: 1655-1658, 1988. PubMed: 2903176

J. Trop. Med. Hyg. 33: 160, 1930.

Chung DI, et al. Biochemical and molecular characterization of a strain KA/S2 of Acanthamoeba castellanii isolated from Korean soil. Korean J. Parasitol. 34: 79-85, 1996. PubMed: 8820744

Gautom RK, et al. Mitochondrial DNA fingerprinting of Acanthamoeba spp. isolated from clinical and environmental sources. J. Clin. Microbiol. 32: 1070-1073, 1994. PubMed: 7913095

Castellani SA. An amoeba found in cultures of a yeast: third note. J. Trop. Med. Hyg. 23: 221-222, 1920.

Ledee DR, et al. Acanthamoeba griffini, molecular characterization of a new corneal pathogen. Invest. Ophthalmol. Vis. Sci. 37: 544-550, 1996. PubMed: 8595954

Castellani A. Maintenance and cultivation of the common pathogenic fungi of man in sterile distilled water; further researches. J. Trop. Med. Hyg. 70: 181-184, 1967.

Van Rooven CE. Observations on the clearing effect of amoeba (Hartmannella castellani) on bacterial cultures: a phenomenon simulating bacteriophagy. J. Trop. Med. Hyg. : 1932..

Daggett PM, et al. Distribution and possible interrelationships of pathogenic and nonpathogenic Acanthamoeba from aquatic environments. Microb. Ecol. 8: 371-386, 1982.

Griffiths AJ, Hughes DE. Starvation and encystment of a soil amoeba Hartmannella castellanii. J. Protozool. 15: 673-677, 1968. PubMed: 5719063

Morton LD, et al. Adherence characteristics of three strains of Acanthamoeba. Rev. Infect. Dis. 13 suppl.5: S424, 1991. PubMed: 2047684

Daggett PM, et al. A molecular approach to the phylogeny of Acanthamoeba. Biosystems 18: 399-405, 1985. PubMed: 4084681

Fritsche TR, et al. Occurrence of bacterial endosymbionts in Acanthamoeba spp. isolated from corneal and environmental specimens and contact lenses. J. Clin. Microbiol. 31: 1122-1126, 1993. PubMed: 8501212

Ott M, et alTemperature-dependent replication of virulent and avirulent Legionella pneumophila isolates in Acanthameoba castellaniiIn: Ott M, et alLegionella: current status and emerging perspectivesWashington, DCAmerican Society for Microbiologypp. 149-151, 1993

Howe D, et al. Identification of two genetic markers that distinguish pathogenic and nonpathogenic strains of Acanthamoeba spp.. Parasitol. Res. 83: 435-348, 1997. PubMed: 9197389

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 H, Chung D. Bacterial endosymbionts of Acanthamoeba sp. isolated from cooling tower water. Jpn. J. Parasitol. 45: 505-511, 1996.

Luck PC, et al. Subinhibitory concentrations of antimicrobial agents reduce the uptake of Legionella pneumophila into Acanthamoeba castellanii and U937 cells by altering the expression of virulence-associated antigens. Antimicrob. Agents Chemother. 42: 2870-2876, 1998. PubMed: 9797218

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

Alves JMP, et al. Random amplified polymorphic DNA probes as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba. Braz. J. Med. Biol. Res. 33: 19-26, 2000.

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

Yagita K, et al. Clustering of Acanthamoeba isolates from human eye infections by means of mitochondrial DNA digestion patterns. Parasitol. Res. 85: 284-289, 1999. PubMed: 10099009

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation