ATCC? Number:30459?
Organism: Entamoeba histolytica Schaudinn
Designations: HM-1:IMSS [ABRM]
Isolation: colonic biopsy of rectal ulcer from adult human male with amebic dysentery, Mexico City, Mexico, 1967
Depositors: LS Diamond
History: ATCC<<--LS Diamond<<--B. Sepulveda/M. de la Torre
Biosafety Level:2
Shipped: frozen
Growth Conditions: ATCCmedium 2463: LYI-S-2 medium, modified
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.
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Comments: Zymodeme II. Mycoplasma-negative by PCR. Viruses [4648] Entamoeba phylogeny [21376] Characterization of glycogen and amino acid pool [23871] Small subunit ribosomal RNA [23923] Phospholipase A enzymes [24140] Effect of growth conditions on isoenzyme patterns [24147] Effect of laminin on amebic pathogenesis [24152] Effect of antagonist of calcium and phospholipase A on the cytopathogenecity [24159] Interactions with polarized human intestinal Caco-2 epithelial cells [24176] High-affinity Gal/GalNAc-specific binding in vitro [42666] Genome sequencing strain (TIGR, The Sanger Institute)
Cross References: Nucleotide (GenBank) : U67158 Entamoeba histolytica 30 kDa type I collagen binding protein mRNA, partial cds.Nucleotide (GenBank) : AAFB02000000Entamoeba histolytica HM-1:IMSS, whole genome shotgun sequencing project.Nucleotide (GenBank) : X98567 E. histolytica ubi1 gene.Nucleotide (GenBank) : M77240 Entamoeba histolytica EHZc3 protein gene, complete cds.Nucleotide (GenBank) : M80910 Entamoeba histolytica serine-rich protein (SREHP) gene, complete cds.Nucleotide (GenBank) : U67157 Entamoeba histolytica 30 kDa type I collagen binding protein mRNA, partial cds.
Classification: KINGDOM: Protozoa
References: 4648: Diamond LS, et al. Viruses of Entamoeba histolytica. I. Identification of transmissible virus-like agents. J. Virol. 9: 326-341, 1972. PubMed: 4335522 5077: Landa L, et al. Advances in methods of Entamoeba histolytica culture. Arch. Invest. Med. 1 suppl.: 9-14, 1970. 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 23871: Bakker-Grunwald T, et al. Characterization of glycogen and amino acid pool of Entamoeba histolytica by C-NMR spectroscopy. J. Eukaryot. Microbiol. 42: 346-349, 1995. PubMed: 7620458 23923: Que X, Reed SL. Nucleotide sequence of a small subunit ribosomal RNA (16S-like rRNA) gene from Entamoeba histolytica: differentiation of pathogenic from nonpathogenic isolates. Nucleic Acids Res. 19: 5438, 1991. PubMed: 1923831 24044: Reed SL, et al. Thiol proteinase expression and pathogenicity of Entamoeba histolytica. J. Clin. Microbiol. 27: 2772-2777, 1989. PubMed: 2556432 24140: Long-Krug SA, et al. Phospholipase A enzymes of Entamoeba histolytica: description and subcellular localization. J. Infect. Dis. 152: 536-541, 1985. PubMed: 2863316 24147: Mirelman D, et al. Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence. Exp. Parasitol. 62: 142-148, 1986. PubMed: 2873049 24152: Li E, et al. Interaction of laminin with Entamoeba histolytica cysteine proteinases and its effect on amebic pathogenesis. Infect. Immun. 63: 4150-4153, 1995. PubMed: 7558332 24159: Ravdin JI, et al. Effect of antagonists of calcium and phospholipase A on the cytopathogenicity of Entamoeba histolytica. J. Infect. Dis. 152: 542-549, 1985. PubMed: 2863317 24176: Li E, et al. Entamoeba histolytica interactions with polarized human intestinal Caco-2 epithelial cells. Infect. Immun. 62: 5112-5119, 1994. PubMed: 7927794 24217: Torian BE, et al. The 96-kilodalton antigen as an integral membrane protein in pathogenic Entamoeba histolytica: potential differences in pathogenic and nonpathogenic isolates. Infect. Immun. 58: 753-760, 1990. PubMed: 2307518 24221: Reed SL, et al. Molecular and cellular characterization of the 29-kilodalton peripheral membrane protein of Entamoeba histolytica: differentiation between pathogenic and nonpathogenic isolates. Infect. Immun. 60: 542-549, 1992. PubMed: 1730488 24236: Mattern CF, et al. Entamoeba histolytica 'toxin' fetuin neutralizable and lectin-like. Am. J. Trop. Med. Hyg. 29: 26-30, 1980. PubMed: 6153255 32946: Flores BM, et al. Surface localization, regulation, and biologic properties of the 96-kDa alcohol/aldehyde dehydrogenase (EhADH2) of pathogenic Entamoeba histolytica. J. Infect. Dis. 173: 226-231, 1996. PubMed: 8537663 33579: Sanchez LB. Cloning and heterologous expression of Entamoeba histolytica adenylate kinase and uridylate/cytidylate kinase. Gene 209: 219-228, 1998. PubMed: 9524270 42666: Pillai DR, et al. The cysteine-rich region of the Entamoeba histolytica adherence lectin (170-kilodalton subunit) is sufficient for high-affinity Gal/GalNAc-specific binding in vitro. Infect. Immun. 67: 3836-3841, 1999. PubMed: 10417146 43275: del Aguila C, et al. Identification of Enterocytozoon bieneusi spores in respiratory samples from an AIDS patient with a 2-year history of intestinal microsporidiosis. J. Clin. Microbiol. 35: 1862-1866, 1997. PubMed: 9196210 70372: Adagu IS, et al. In vitro activity of nitazoxanide and related compounds against isolates of Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. J. Antimicrob. Chemother. 49: 103-111, 2002. PubMed: 11751773 70542: Sanchez LB, et al. Fructose-1,6-bisphosphate aldolases in amitochondriate protists constitute a single protein subfamily with eubacterial relationships. Gene 295: 51-59, 2002. PubMed: 12242011
