Trichomonas vaginalis Donne (ATCC® PRA-98)

Organism: Trichomonas vaginalis Donne  /  Depositor: PJ Johnson

Permits and Restrictions

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Strain Designations G3
Application
Sexually Transmitted Disease Research
Biosafety Level 2
Isolation
Clinical specimen - human, Beckenham United Kingdom, 1973
Product Format frozen
Storage Conditions Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Genome Sequenced Strain

Yes

Comments
Genome sequencing strain
In vitro susceptibility to metronidazole
Medium ATCC® Medium 2154: LYI Entamoeba medium
ATCC® Medium 361: Modified TYM basal medium (ATCC medium 358) with pH adjusted to 6.0 and 0.2-0.5 ml of heat-inactivated horse serum added per tube before use
Growth Conditions Temperature: 35°C
Atmosphere:  Anaerobic
Culture System: Axenic; pH 6
Cryopreservation Harvesting and Preservation
  1. Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min. The cells grown in a medium containing agar are concentrated by centrifugation, a solid pellet does not form. The soft pellet is resuspended to desired cell concentration with agar-free supernatant.
  2. Adjust the concentration of cells to 2 x 10to  2 x 107/mL in fresh medium.
  3. While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium.
    1. Add 1.0 mL of DMSO to an ice cold 20 x 150 mm screw-capped test tube.
    2. Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 9.0 mL of ice cold medium.
    3. Invert several times to dissolve the DMSO.
    4. Allow to warm to room temperature.
  4. Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 106 to  107 cells/mL and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should no less than 15 min and no longer than 30 min.
  5. Dispense in 0.5 mL aliquots into 1.0 mL to 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
  6. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion.  At -40°C plunge 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. The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials should not be stored above -55°C.
  8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial just to a level just above the surface of 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 a 16 x 125 mm screw-capped test tube containing either 9 mL of ATCC medium 361 (completed with serum) or 13 mL ATCC Medium 2154 adjusted to pH 6.0.
  10. Incubate the culture at 35°C with the cap screwed on tightly (tube should be vertical for medium 361 or on a 15° horizontal slant for medium 2154).
Name of Depositor PJ Johnson
Special Collection NSF - Protistology
Chain of Custody
P J Johnson
G H Coombs
Year of Origin 1973
References

Bremner AF, et al. Antitrichomonal activity of alpha-difluoromethylornithine. J. Antimicrob. Chemother. 20: 405-411, 1987. PubMed: 3119554

Clackson TE, Coombs GH. The antagonistic effects of acetate and lactate upon the trichomonacidal activity of metronidazole. J. Antimicrob. Chemother. 11: 401-406, 1983. PubMed: 6603456

Coombs GH, North MJ. An analysis of the proteinases of Trichomonas vaginalis by polyacrylamide gel electrophoresis. Parasitology 86: 1-6, 1983. PubMed: 6340036

Coombs GH, Clackson TE. Antitrichomonal activity of compounds that affect DNA and its repair. J. Antimicrob. Chemother. 11: 191-194, 1983. PubMed: 6601103

Coombs GH, Mottram JC. Trifluoromethionine, a prodrug designed against methionine gamma-lyase-containing pathogens, has efficacy in vitro and in vivo against Trichomonas vaginalis. Antimicrob. Agents Chemother. 25: 1743-1745, 2001. PubMed: 11353620

Cornelius DC, et al. Short report: genetic relatedness of Trichomonas vaginalis reference and clinical isolates. Am. J. Trop. Med. Hyg. 83: 1283-1286, 2010.

Hassan HF, Coombs GH. Phosphomonoesterases of Leishmania mexicana mexicana and other flagellates. Mol. Biochem. Parasitol. 23: 285-296, 1987. PubMed: 3037369

Hirt RP, et al. Biochemical and genetic evidence for a family of heterotrimeric G-proteins in Trichomonas vaginalis. Mol. Biochem. Parasitol. 129: 179-189, 2003. PubMed: 12850262

Irvine JW, et al. Purification of cysteine proteinases from trichomonads using bacitracin-Sepharose. FEMS Microbiol. Lett. 110: 113-119, 1993. PubMed: 8319888

Irvine JW, et al. Use of inhibitors to identify essential cysteine proteinases of Trichomonas vaginalis. FEMS Microbiol. Lett. 149: 45-50, 1997. PubMed: 9103977

Lau AO, et al. Trichomonas vaginalis initiator binding protein, IBP39, contains a novel DNA binding motif. Mol. Biochem. Parasitol. 130: 167-171, 2003. PubMed: 12946857

Lockwood BC, Coombs GH. Purification and characterization of methionine gamma-lyase from Trichomonas vaginalis. Biochem. J. 279: 675-682, 1991. PubMed: 1953661

Lockwood BC, et al. Trichomonas vaginalis, Tritrichomonas foetus, and Trichomitus batrachorum: comparative proteolytic activity. Exp. Parasitol. 58: 245-253, 1984. PubMed: 6389167

Lockwood BC, et al. The release of hydrolases from Trichomonas vaginalis and Tritrichomonas foetus. Mol. Biochem. Parasitol. 30: 135-142, 1988. PubMed: 3140008

Lockwood BC, et al. The use of a highly sensitive electrophoretic method to compare the proteinases of trichomonads. Mol. Biochem. Parasitol. 24: 89-95, 1987. PubMed: 3302703

Mallinson DJ, et al. Identification and molecular cloning of four cysteine proteinase genes from the pathogenic protozoon Trichomonas vaginalis. Microbiology 140: 2725-2735, 1994. PubMed: 8000542

McKie AE, et al. The primitive protozoon Trichomonas vaginalis contains two methionine gamma-lyase genes that encode members of the gamma-family of pyridoxal 5'-phosphate-dependent enzymes.. J. Biol. Chem. 273: 5549-5556, 1998. PubMed: 9488680

North MJ, et al. Polyamine biosynthesis in trichomonads. Mol. Biochem. Parasitol. 19: 241-249, 1986. PubMed: 3090433

North MJ, et al. The specificity of trichomonad cysteine proteinases analysed using fluorogenic substrates and specific inhibitors. Mol. Biochem. Parasitol. 39: 183-193, 1990. PubMed: 2320056

Thong KW, Coombs GH. Comparative study of ferredoxin-linked and oxygen-metabolizing enzymes of trichomonads. Comp. Biochem. Physiol., B 87: 637-641, 1987. PubMed: 3497772

Thong KW, Coombs GH. The effects of inhibitors of sulphur-containing amino acid metabolism on the growth of Trichomonas vaginalis in vitro. J. Antimicrob. Chemother. 19: 429-437, 1987. PubMed: 3495529

Thong KW, et al. L-methionine catabolism in trichomonads. Mol. Biochem. Parasitol. 23: 223-231, 1987. PubMed: 3496535

Thong KW, et al. S-Adenosylhomocysteine hydrolase activity in Trichomonas vaginalis and other trichomonads. Mol. Biochem. Parasitol. 17: 35-44, 1985. PubMed: 3932851

Thong KW, et al. S-adenosylmethionine and transmethylation reactions in trichomonads. Parasitol. Res. 73: 193-198, 1987. PubMed: 3108875

Thong KW, Coombs GH. Trichomonas species: homocysteine desulphurase and serine sulphydrase activities. Exp. Parasitol. 63: 143-151, 1987. PubMed: 3494628

Scott DA, et al. Trichomonas vaginalis: amoeboid and flagellated forms synthesize similar proteinases. Exp. Parasitol. 80: 345-348, 1995. PubMed: 7895847

Scott DA, et al. The pathway of secretion of proteinases in Trichomonas vaginalis. Int. J. Parasitol. 25: 657-666, 1995. PubMed: 7657450

Zuo X, et al. Uptake of amino acids by the parasitic, flagellated protist Trichomonas vaginalis. Microbiology 141: 2637-2642, 1995. PubMed: 7582024

Cross References

Nucleotide (GenBank) : AAHC01000000 Trichomonas vaginalis G3, whole genome shotgun sequencing project.

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.
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