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ATCC? Number:30015?
Organism: Entamoeba histolytica Schaudinn
Designations: HK-9
Isolation: colonic biopsy from adult human male with amebic dysentery, Korea, (?)
Depositors: LS Diamond
Biosafety Level:2
Shipped: frozen
Growth Conditions: ATCC medium 2154: LYI Entamoeba medium

Temperature: 35.0°C 

Duration: axenic; anaerobic Protocol: The following directions for recovery of this strain from the frozen state must be carefully followed if a culture is to be established successfully.
1. Warm a fresh tube of the appropriate medium (ATCCmedium 2154 [available from ATCCas item PRA-2154 LYI Entamoeba Medium] or, for those strains that require extra serum, ATCCmedium 2154 plus 15% heat-inactivated bovine serum) to 35C.
2. Place the frozen ampule in a water bath at 35C. Allow the ampule to remain in the water bath until completely thawed (about 2 minutes). NOTE: If the frozen ampule cannot be thawed immediately upon arrival, store it at -130C or lower.
3. Transfer the thawed material to the fresh, warmed tube of medium, cap tightly, and gently invert the tube several times.
4. Place the tube on a 15-degree slant in a 35C incubator and allow it to remain undisturbed, except to observe it daily with an inverted microscope.
5. After 2 or 3 days if only a few cells are attached, centrifuge the tube at 200 x g for 5 minutes, remove all except about 1.0 ml of the supernatant, and add fresh medium. Return the culture to a 35C incubator and monitor daily.
6. When the culture forms a monolayer on the bottom surface of the slanted tube, use the following protocol to transfer it: Ice the culture for 10 minutes, invert it 20 times, and aseptically transfer 0.1-0.3 ml to a fresh tube of the appropriate medium. Incubate as above. The transfer interval will vary with the quality of the medium, but is generally between 3 and 4 days.
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications: control strain  [92352] media testing
Comments: Entamoeba phylogeny  [21376] Zymodeme II not detected by Trichomonas vaginalis-specific PCR primers  [37722] Axenic cultivation  [5053] use of plastic ampoules for freeze preservation  [23870] Growth responses to reducing agents  [24041] Effect of cysteine and oxygen tension on survival  [24185] requirement for cysteine and ascorbic acid for attachment  [24161]
Classification: KINGDOM: Protozoa
References:  4903: Diamond LS. Axenic cultivation of Entamoeba histolytica.  Science 134: 336-337, 1961. PubMed: 13722605 

5053: Diamond LS. Techniques of axenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae.  J. Parasitol. 54: 1047-1056, 1968. PubMed: 4319346 

21376: Clark CG, Diamond LS. Intraspecific variation and phylogenetic relationships in the genus Entamoeba as revealed by riboprinting.  J. Eukaryot. Microbiol. 44: 142-154, 1997. PubMed: 9109261 

23870: Simione FP Jr., et al. The use of plastic ampoules for freeze preservation of microorganisms.  Cryobiology 14: 500-502, 1977. PubMed: 891238 

23954: Gonzalez-Ruiz A, et al. Diagnosis of amebic dysentery by detection of Entamoeba histolytica fecal antigen by an invasive strain-specific, monoclonal antibody-based enzyme-linked immunosorbent assay.  J. Clin. Microbiol. 32: 964-970, 1994. PubMed: 8027351 

23988: Charoenlarp P, et al. Carbohydrate utilization by Entamoeba histolytica.  Exp. Parasitol. 23: 205-211, 1968. PubMed: 4302387 

24015: Takeuchi T, et al. Pyruvate oxidase (CoA acetylating) in Entamoeba histolytica.  Biochem. Biophys. Res. Commun. 65: 591-596, 1975. PubMed: 167776 

24041: Gillin FD, Diamond LS. Entamoeba histolytica and Giardia lamblia: growth responses to reducing agents.  Exp. Parasitol. 51: 382-391, 1981. PubMed: 6262103 

24133: Takeuchi T, et al. Entamoeba histolytica: localization and characterization of phosphorylase and particulate glycogen.  Exp. Parasitol. 43: 107-114, 1977. PubMed: 196880 

24161: Gillin FD, Diamond LS. Attachment of Entamoeba histolytica to glass in a defined maintenance medium: Specific requirement for cysteine and ascorbic acid.  J. Protozool. 27: 474-478, 1980. PubMed: 6260930 

24185: Gillin FD, Diamond LS. Entamoeba histolytica and Giardia lamblia: effects of cysteine and oxygen tension on trophozoite attachment to glass and survival in culture media.  Exp. Parasitol. 52: 9-17, 1981. PubMed: 6263667 

24203: Gelderman AH, et al. A comparison of genome sizes and thermal-denaturation-derived base composition of DNAs from several members of Entamoeba (histolytica group).  J. Parasitol. 57: 912-916, 1971. PubMed: 4327967 

24204: Phillips BP, et al. Results of intracecal inoculation of germfree and conventional guinea pigs and germfree rats with axenically cultivated Entamoeba histolytica.  J. Protozool. 19: 498-499, 1972. PubMed: 4342299 

24207: Garcia LS, et al. Comparison of indirect fluorescent-antibody amoebic serology with counterimmunoelectrophoresis and indirect hemagglutination amoebic serologies.  J. Clin. Microbiol. 15: 603-605, 1982. PubMed: 6279694 

24208: Gelderman AH, et al. Characterization of the deoxyribonucleic acid of representative strains of Entamoeba histolytica, E. histoytica-like amebae, and E. moshkovskii.  J. Parasitol. 57: 906-911, 1971. PubMed: 4327966 

37722: Madico G, et al. Diagnosis of Trichomonas vaginalis infection by PCR using vaginal swab samples.  J. Clin. Microbiol. 36: 3205-3210, 1998. PubMed: 9774566 

92352: Procedures for the Recovery and Identification of Parasites from the Intestinal Tract: Approved Guideline - 2nd Edition. Wayne, PA:Clinical and Laboratory Standards Institute;CLSI M28-A2.