Trichomonas vaginalis Donne

30001 ^™

Trichomonas vaginalis strain C-1:NIH was isolated from the vaginal exudate of a female with acute vaginitis. This parasitic protozoan strain produces glucokinase and ketohexokinase fructokinase.

Product category: Protists
Product type: Parasitic protozoan
Classification: Eukaryota, Metamonada, Parabasalia, Trichomonadida, Trichomonadidae, Trichomonas
Strain designation: C-1:NIH
Type strain: No
Isolation source: Vaginal exudate from female with acute vaginitis
Applications: Infectious disease research

Sexually transmitted disease research
Product format: Frozen
Storage conditions: -80°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage

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Documentation

Product sheet

BSL 2

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

LYI Entamoeba Medium

PRA-2154

General

Specific applications: Produces glucokinase
Produces ketohexokinase fructokinase
Maltose utilization

Characteristics

Comments: Phylogeny based upon superoxide dismutase gene sequence analysis
Diagnosis of Trichomonas vaginalis by PCR methods
Inorganic pyrophosphatase
Hydrogenosomal succinate thiokinase
Glucokinase and fructokinase
Differences in strains virulence
Maltose utilization
Carbon metabolism based on carbon source
Carbohydrate metabolism in chemostats
Antioxidant defenses
Metronidazole radical anion generation
Primary structure of the pyruvate:ferredoxin oxidoreductase
Primary structure of the hydrogenosomal malic enzyme
Response of lymphocytes
Fructose-2,6-bisphosphate-insensitive pyrophosphate:fructose-6-phosphate phosphotransferase
A linear double-stranded RNA
Viability at four temperatures
Extranuclear DNA
Effect of oxygen and carbon dioxide on growth
Virus RNAs
Properties of a secondary alcohol dehydrogenase
Cytochemistry of hydrogenosomal enzymes
Ferredoxin-dependent reduction of nitroimidazole derivatives
Subcellular localization of enzymes of the arginine dihydrolase pathway
In vitro susceptibility to metronidazole

Handling information

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

Instruction for complete medium

Media: ATCC Medium 2154 adjusted to pH 6.0 with the addition 150 µL sterile 1N HCl per 13 mL of medium.

Alternate Media: ATCC Medium 361

Temperature

35°C

Atmosphere

Microaerophilic

Culture system

Axenic

Handling procedure

Storage and Culture Initiation

Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen. If liquid nitrogen storage facilities are not available, frozen ampules may be stored at or below -70°C for approximately one week. Do not under any circumstance store frozen ampules at refrigerator freezer temperatures (generally -20°C). Storage of frozen material at this temperature will result in the death of the culture.

To thaw a frozen ampule, place it in a 35°C water bath , until thawed (2-3 min). Immerse the ampule just sufficient to cover the frozen material. Do not agitate the ampule.
Immediately after thawing, aseptically transfer contents to a 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. Incubate the tube at 35°C (tube should be vertical for medium 361 or on a 15° horizontal slant for medium 2154).

Handling notes

Molecular authentication of the source material (seed stock) for distribution lots of ATCC 30001 has been performed at the species level by sequencing of the Trichomonas vaginalis SSU rRNA and AP65 adhesin genes.

Culture maintenance

When the culture is at or near peak density, place the tubes on ice for 10 minutes.
Gently invert the culture tube 10 times and aseptically transfer a 0.1-0.4 mL aliquot to a 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.
Incubate the culture at 35°C (tube should be vertical for medium 361 or on a 15º horizontal slant for medium 2154).
Transfer the culture every 3-4 days as described in steps 1-2. The transfer interval will depend on the quantity of the inoculum and the quality of the medium. This should be empirically determined by examining the culture on a daily basis until the growth cycle has stabilized. Do not allow the culture to overgrow. The culture crashes soon after reaching peak density.

Cryopreservation

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.
Adjust the concentration of cells to 2 x 10⁶- 2 x 10⁷/mL in fresh medium.
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.
Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 10⁶ - 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.
Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
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.)
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.
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.
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.
Incubate the culture at 35ºC with the cap screwed on tightly (tube should be vertical for medium 361 or on a 15º horizontal slant for medium 2154).

History

Deposited as: Trichomonas vaginalis Donne
Depositors: LS Diamond
Chain of custody: ATCC <-- LS Diamond <-- TA Burch/L.V. Reardon <-- L Jacobs
Type of isolate: Human
Year of origin: 1956
Patient age: adult
Patient gender: Female
Special collection: NCRR Contract
Cross references: GenBank Z70670 T.vaginalis sod1 gene.

GenBank Z70671 T.vaginalis sod2 gene.

GenBank Z70672 T.vaginalis sod3 gene.

GenBank Z70673 T.vaginalis sod4 gene.

GenBank Z70674 T.vaginalis sod5 gene.

GenBank M33717 T.vaginalis ferredoxin gene, complete cds.

GenBank U57000 chaperonin 60 (cpn60) mRNA, partial coding sequence

GenBank AF067404 DNA polymerase epsilon gene, partial coding sequence

GenBank X98016 histone H4-2 and histone H3-2, partial coding sequence

GenBank U27577 Trichomonas vaginalis polyubiquitin (UbA) mRNA, partial cds.

GenBank U28008 Trichomonas vaginalis ubiquitin 1A (Ub1A) gene, partial cds.

GenBank U28009 Trichomonas vaginalis ubiquitin 1C (Ub1C) gene, partial cds.

GenBank U28010 Trichomonas vaginalis ubiquitin 1D (Ub1D) gene, partial cds.

GenBank U28011 Trichomonas vaginalis ubiquitin 1E (Ub1E) gene, partial cds.

GenBank U28012 Trichomonas vaginalis ubiquitin dimer 2B (Ub2B) gene, partial cds.

GenBank U70308 Trichomonas vaginalis mitochondrial-type HSP70 mRNA, complete cds.

GenBank U38692 Trichomonas vaginalis cytosolic malate dehydrogenase gene, complete cds.

GenBank U28013 Trichomonas vaginalis polyubiquitin junction JC (UbJC) gene, partial cds.

GenBank AF060233 Trichomonas vaginalis L-lactate dehydrogenase (LDH1) gene, complete cds.

GenBank AF058282 Trichomonas vaginalis elongation factor 1 alpha (tef1) mRNA, partial cds.

GenBank L11394 Trichomonas vaginalis glyceraldehyde 3-phosphate dehydrogenase mRNA, 3' end.

GenBank U07784 Trichomonas vaginalis ATCC 30001 ferredoxin (FD) gene, Inr promoter element.

GenBank M97553 Trichomonas vaginalis succinyl-CoA synthetase beta-subunit gene, complete cds.

GenBank U07785 Trichomonas vaginalis ATCC 30001 P-glycoprotein (Pgp1) gene, promoter element.

GenBank AF022421 glyceraldehyde-3-phosphate dehydrogenase (gap3) gene, partial coding sequence

GenBank U07782 Trichomonas vaginalis ATCC 30001 beta-tubulin (beta-Tub) gene, promoter element.

GenBank U07780 Trichomonas vaginalis ATCC 30001 alpha-tubulin (alpha-Tub) gene, promoter element.

GenBank U07203 Trichomonas vaginalis hydrogenosomal adenylate kinase proprotein gene, complete cds.

GenBank U16836 Trichomonas vaginalis hydrogenosomal malic enzyme subunit A (maeA) gene, complete cds.

GenBank AF022414 Trichomonas vaginalis glyceraldehyde-3-phosphate dehydrogenase (gap2) gene, partial cds.

GenBank AF005075 translation initiation factor 2 gamma subunit (eIF-2 gamma) gene, partial coding sequence

GenBank U38786 calmodulin (CAM) and E2 ubiquitin-conjugating enzyme (TvUBC) genes, partial coding sequence

GenBank AF053370 PPi-dependent fructose 6-phosphate 1-phosphotransferase (pfk3) gene, partial coding sequence

GenBank U16822 Trichomonas vaginalis pyruvate:ferredoxin oxidoreductase proprotein (pfoA) gene, complete cds.

GenBank U16823 Trichomonas vaginalis pyruvate:ferredoxin oxidoreductase proprotein (pfoB) gene, complete cds.

GenBank U16838 Trichomonas vaginalis hydrogenosomal malic enzyme subunit C proprotein (maeC) gene, partial cds.

GenBank U16839 Trichomonas vaginalis hydrogenosomal malic enzyme subunit D proprotein (maeD) gene, partial cds.

GenBank U16837 Trichomonas vaginalis hydrogenosomal malic enzyme subunit B proprotein (maeB) gene, complete cds.

GenBank U07783 Trichomonas vaginalis ATCC 30001 70kDa cystolic heat shock protein (cHSP70 gene), promoter element.

GenBank AF044973 pyrophosphate-dependent fructose 6-phosphate 1-phosphotransferase (Pfk1) gene, complete coding sequence

GenBank AF053371 Trichomonas vaginalis PPi-dependent fructose 6-phosphate 1-phosphotransferase (pfk2) gene, partial cds.

GenBank U07781 Trichomonas vaginalis ATCC 30001 succinyl CoA synthetase beta subunit (beta-SCS) gene, Inr promoter element.

GenBank U07779 Trichomonas vaginalis ATCC 30001 succinyl CoA synthetase alpha subunit (alpha-SCSB) gene, Inr promoter element.

Legal disclaimers

Intended use

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

Warranty

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Searle SM, Muller M. Inorganic pyrophosphatase of Trichomonas vaginalis. Mol. Biochem. Parasitol. 44: 91-96, 1991. PubMed: 1849232

Lloyd D, Pedersen JZ. Metronidazole radical anion generation in vivo in Trichomonas vaginalis: oxygen quenching is enhanced in a drug-resistant strain. J. Gen. Microbiol. 131: 87-92, 1985. PubMed: 2985740

Jenkins TM, et al. Hydrogenosomal succinate thiokinase in Tritrichomonas foetus and Trichomonas vaginalis. Biochem. Biophys. Res. Commun. 179: 892-896, 1991. PubMed: 1898409

Mertens E, Muller M. Glucokinase and fructokinase of Trichomonas vaginalis and Tritrichomonas foetus. J. Protozool. 37: 384-388, 1990. PubMed: 2213652

Reardon LV, et al. Differences in strains of Trichomonas vaginalis as revealed by intraperitoneal injections into mice. J. Parasitol. 47: 527-532, 1961. PubMed: 13740097

View All Curated Citations for this Product

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