Naegleria fowleri Carter (ATCC® 30894)

Strain Designations: Lee (L.L.)  /  Depositor: DT John  /  Biosafety Level: 2

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Strain Designations Lee (L.L.)
Biosafety Level 2

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 Cerebrospinal fluid of 15-year-old female, Richmond, VA, 1968
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 no
Arginine-dependent cytolytic mechanicm
Resistence to complement-mediated lysis
Innate resistance of mice
Immunization of mice
Alterations in protein expression and complement resistance
Varying the serum component in axenic cultivation
DNA fingerprinting
biochemical identification
Multicomponent hemolytic system
Membrane vesiculation for resisting complement damage
Modulation of virulence by alterations in growth media
Interrepeat PCR
Transmission between mice
Subcellular distribution of hydrolases
Thermal ecology
Serum agglutination and immunoglobulin levels in infected mice
Infection acquired by mice through swimming
Comparison of two species cultivated in the same nutrient medium
Method for assessing the migratory response
Differentiation of Naegleria fowleri from Acanthamoeba using monocolonal antibody
Medium ATCC® Medium 1034: Modified PYNFH medium (Available from ATCC as ATCC cat. no. 327-X)
ATCC® Medium 710: Nelson's Culture Medium For Naegleria
ATCC® Medium 803: M7 medium
ATCC® Medium 902: Schuster's axenic Naegleria medium
Growth Conditions
Temperature: 35°C
Culture System: Axenic
Cryopreservation Harvest and Preservation
  1. Harvest cells from a culture that is at or near peak density by centrifugation at 600 x g for 5 min. Pool the cell pellets into a single tube.
  2. Adjust the concentration of cells to 2.0 x 106/mL.  If the concentration is too low, centrifuge at 600 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired concentration.
  3. Prepare a 15% (v/v) sterile DMSO solution in ATCC medium 1034 as follows:  Add the required volume of DMSO to a glass screw-capped test tube and place on ice.  Allow the DMSO to solidify.  Add the required volume of refrigerated ATCC medium 1034.  Dissolve the DMSO by inverting several times.  If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
  4. Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 106 and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 60 min.
  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 the ampules into liquid nitrogen.
  7. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen refrigerator.
  8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial enough to cover only the frozen material. Do not agitate the vial.
  9. Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate into 5.0 mL of fresh ATCC medium 1034.
  10. Incubate the tube on a 15° horizontal at 35°C with the cap screwed on tightly.
Mycoplasma Unknown
Name of Depositor DT John
Special Collection NCRR Contract
Chain of Custody
ATCC <-- DT John <-- E.C. Nelson
Year of Origin 1968

Haggerty RM, John DT. Innate resistance of mice to experimental infection with Naegleria fowleri. Infect. Immun. 20: 73-77, 1978. PubMed: 669800

John DT, et al. Immunization of mice against Naegleria fowleri infection. Infect. Immun. 16: 817-820, 1977. PubMed: 892900

Daggett PM, Nerad TA. The biochemical identification of vahlkampfiid amoebae. J. Protozool. 30: 126-128, 1983. PubMed: 6864593

Marciano-Cabral F, et al. Cytopathic action of Naegleria fowleri amoebae on rat neuroblastoma target cells. J. Protozool. 37: 138-144, 1990. PubMed: 2108243

Haight JB, John DT. Varying the serum component in axenic cultures of Naegleria fowleri. Proc. Helminthol. Soc. Wash. 49: 127-134, 1982.

Whiteman LY, Marciano-Cabral F. Resistance of highly pathogenic Naegleria fowleri amoebae to complement-mediated lysis. Infect. Immun. 57: 3869-3875, 1989. PubMed: 2807551

Duma RJ, et al. Primary amebic meningoencephalitis. N. Engl. J. Med. 281: 1315-1323, 1969. PubMed: 5355436

Toney DM, Marciano-Cabral F. Alterations in protein expression and complement resistance of pathogenic Naegleria amoebae. Infect. Immun. 60: 2784-2790, 1992. PubMed: 1319405

van Belkum A. DNA fingerprinting of medically important microorganisms by use of PCR. Clin. Microbiol. Rev. 7: 174-184, 1994. PubMed: 8055466

Fischer-Stenger K, Marciano-Cabral F. The arginine-dependent cytolytic mechanism plays a role in destruction of Naegleria fowleri amoebae by activated macrophages. Infect. Immun. 60: 5126-5131, 1992. PubMed: 1452346

John DT. Primary amebic meningoencephalitis and the biology of Naegleria fowleri. Annu. Rev. Microbiol. 36: 101-123, 1982. PubMed: 6756287

Weik RR, John DT. Quantitation and cell size of Naegleria fowleri by electronic particle counting. J. Parasitol. 63: 150-151, 1977. PubMed: 321738

Lowrey DM, McLaughlin J. A multicomponent hemolytic system in pathogenic amoebae Naegleria fowleri. Infect. Immun. 45: 731-736, 1984. PubMed: 6469359

Detterline JL, Wilhelm WE. Survey of pathogenic Naegleria fowleri and thermotolerant amebas in Federal Recreational Waters. Trans. Am. Microsc. Soc. 110: 244-261, 1991.

Toney DM, Marciano-Cabral F. Membrane vesiculation of Naegleria fowleri amoebae as a mechanism for resisting complement damage. J. Immunol. 152: 2952-2959, 1994. PubMed: 8144894

Toney DM, Marciano-Cabral F. Modulation of complement resistance and virulence of Naegleria fowleri amoebae by alterations in growth media. J. Eukaryot. Microbiol. 41: 337-343, 1994. PubMed: 8087105

van Belkum A, et al. Genotyping Naegleria spp. and Naegleria Fowleri isolates by Interepeat Polymerase Chain reaction. J. Clin. Microbiol. 30: 2595-2598, 1992. PubMed: 1400959

May RG, John DT. Transmission of Naegleria fowleri between mice. J. Parasitol. 69: 249-251, 1983. PubMed: 6827444

Weik RR, John DT. Agitated mass cultivation of Naegleria fowleri. J. Parasitol. 63: 868-871, 1977. PubMed: 21233

Weik RR, John DT. Cell and mitochondria respiration of Naegleria fowleri. J. Parasitol. 65: 700-708, 1979. PubMed: 41892

Lowrey DM, McLaughlin J. Subcellular distribution of hydrolases in Naegleria fowleri. J. Protozool. 32: 616-621, 1985. PubMed: 2999380

Adams AC, et al. Modification of resistance of mice to Naegleria fowleri infections. Infect. Immun. 13: 1387-1391, 1976. PubMed: 1270145

Weik RR, John DT. Macromolecular composition and nuclear number during growth of Naegleria fowleri. J. Parasitol. 64: 746-747, 1978. PubMed: 682075

Huizinga HW, McLaughlin GL. Thermal ecology of Naegleria fowleri from a power plant cooling reservoir. Appl. Environ. Microbiol. 56: 2200-2205, 1990. PubMed: 1975164

Haggerty RM, John DT. Serum agglutination and immunoglobulin levels of mice infected with Naegleria fowleri. J. Protozool. 29: 117-122, 1982. PubMed: 7086710

John DT, Nussbaum SL. Naegleria fowleri infection acquired by mice through swimming in amebae-contaminated water. J. Parasitol. 69: 871-874, 1983. PubMed: 6672166

Fulford DE, et al. Cytopathogenicity of Naegleria fowleri for cultured rat neuroblastoma cells. J. Protozool. 32: 176-180, 1985. PubMed: 3989747

Cline M, et al. Comparison of Naegleria fowleri and Naegleria gruberi cultivated in the same nutrient medium. J. Protozool. 30: 387-391, 1983. PubMed: 6631780

Fischer-Stenger K, et al. Separation of soluble amoebicidal and tumoricidal activity of activated macrophages. J. Protozool. 39: 235-241, 1992. PubMed: 1560419

Brinkley C, Marciano-Cabral F. A method for assessing the migratory response of Naegleria fowleri utilizing [3H]uridine-labeled amoebae. J. Protozool. 39: 297-303, 1992. PubMed: 1578403

Flores BM, et al. Differentiation of Naegleria fowleri from Acanthamoeba species by using monoclonal antibodies and flow cytometry. J. Clin. Microbiol. 28: 1999-2005, 1990. PubMed: 2229384

Alizadeh H, et al. Tear IgA and serum IgG antibodies against Acanthamoeba in patients with Acanthamoeba keratitis. Cornea 20: 622-627, 2001. PubMed: 11473164

Herbst R, et al. Pore-forming polypeptides of the pathogenic protozoon Naegleria fowleri. J. Biol. Chem. 277: 22353-22360, 2002. PubMed: 11948186

Reveiller FL, et al. Species specificity of a monoclonal antibody produced to Naegleria fowleri and partial characterization of its antigenic determinant. Parasitol. Res. 86: 634-641, 2000. PubMed: 10952262

Barbour SE, Marciano-Cabral F. Naegleria fowleri amoebae express a membrane-associated calcium-independent phospholipase A(2). Biochim. Biophys. Acta 1530: 123-133, 2001. PubMed: 11239815

Reveiller FL, et al. Isolation of a unique membrane protein from Naegleria fowleri. J. Eukaryot. Microbiol. 48: 676-682, 2001. PubMed: 11831777

Chu D-M, et al. Calcium-dependent protection from complement lysis in Naegleria fowleri amebae. Cell Calcium 31: 105-114, 2002. PubMed: 12027384

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