ATCC? Number:50261?
Organism: Entamoeba sp.
Designations: EC
Isolation: sewage, Guayaquil, Ecuador, 1965
Depositors: LS Diamond
History: ATCC<<--LS Diamond<<--R.A. Neal <<-- R. Marcias-Constante
Biosafety Level:1
Shipped: test tube
Growth Conditions: ATCCmedium 1171: TYGM-9 mediumTemperature: 25.0°C Duration: anaerobic; grown with bacteria Protocol: ATCCNO: 30131 SPEC: This culture is xenic; i.e., it contains mixed unidentified bacteria, some or all of which serve as food for the amoeba. 1) A growing culture is shipped in a 16 X 125 mm screw-capped test tube filled to within approximately two centimeters of the top with medium, a configuration which enhances survival in transit. 2) Immediately upon receipt of the culture, place it on a 15-degree slant at 35C. Allow the culture to remain undisturbed for at least three hours. 3) Observe the culture with an inverted microscope. Attached trophozoites should be evident. Reduce the volume of the culture to approximately 9 ml. 4) Centrifuge the removed culture fluid at 500 x g for five minutes. Under these conditions any trophozoites in suspension will be pelleted to the bottom of the tube. 5) Inoculate two fresh tubes of ATCCmedium 1171 (available from ATCCas item IV-1171) with 0.25 ml of the supernatant derived in step 4 and incubate the tubes at 35C. These tubes will serve as preinoculated bacterized culture tubes. Preinoculation of medium with bacteria prior to subcultivation of Dientamoeba and bacterized Entamoeba strains allows for better growth. 6) Divide the remainder of the supernatant from step 4 into two equal aliquots in 16 X 125 mm screw-capped test tubes. Increase the volume of each tube to approximately 9 ml with fresh ATCCmedium 1171. 7) Ice the parent shipped culture for five minutes, invert the tube 20 times and transfer 0.5- and 1.0-ml aliquots to the tubes just set up in step 6. 8) Incubate all cultures at 35C. Transfer cultures when they reach early stationary phase. The transfer interval will depend on the quality of the culture medium used. Inoculate bacterized culture tubes at least one day prior to subcultivation of Dientamoeba and bacterized Entamoeba strains. 9) In general, addition of penicillin G at 75 U/ml and streptomycin at 75 mcg/ml to ATCCmedium 1171 may be necessary if the bacterial density is too high.
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Type Strain: yes(type strain)
Comments: Entamoeba histolytica-like amoebae are Entamoeba moshkovskii [4628]
Classification: KINGDOM: Protozoa
References: 4628: Clark CG, Diamond LS. The Laredo strain and other 'Entamoeba histolytica-like' amoebae are Entamoeba moshkovskii. Mol. Biochem. Parasitol. 46: 11-18, 1991. PubMed: 1677159 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