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Spironucleus vortens Poynton et al.

50386

Product category
Protists
Product type
Parasitic protozoan
Type strain
Yes
Isolation source
lip lesion, angelfish, Pterophyllum scalare, FL (29 degrees 32' N, 82 degrees 23' W)
Geographical isolation
United States; Florida; Archer
Product format
Frozen
Storage conditions
-80°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage
Mission Collection Item
This is a Mission Collection Item

Documentation

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Biosafety Icon BSL 1

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.

Detailed product information

Handling information

Medium
Instruction for complete medium

Medium: ATCC Medium 2154
(ATCC medium 2154 is available in a freeze-dried format as cat. no. PRA-2154; contact ATCC sales for details).

Temperature
35°C
Atmosphere
Anaerobic
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.

  1. 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.
  2. Immediately after thawing, aseptically transfer contents to a screw-capped test tube containing 13 ml ATCC Medium 2154.  Incubate the tube on a 15° horizontal slant at 35°C.
Culture maintenance
  1. When the culture is at or near peak density, place the tubes on ice for 10 minutes. 
  2. Gently invert the culture tube 10 times and aseptically transfer a 0.1-0.4 ml aliquot to a screw-capped test tube containing 13 ml ATCC Medium 2154. 
  3. Incubate the culture on a 15° horizontal slant at 35°C. 
  4. Transfer the culture weekly 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
  1. Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.
  2. Adjust the concentration of cells to 2 x 106 - 2 x 107/ml in fresh medium.
  3. While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium.
    a) Add 1.0 ml of DMSO to an ice cold 20 x 150 mm screw-capped test tube;
    b) Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 9.0 ml of ice cold medium;
    c) Invert several times to dissolve the DMSO;
    d) Allow to warm to room temperature.
  4. 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.
  5. Dispense in 0.5 ml aliquots into 1.0 - 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 13 ml ATCC Medium 2154.
  10. Incubate the culture on a 15° horizontal slant at 35°C with the cap screwed on tightly.

History

Deposited as
Spironucleus vortens Poynton et al.
Depositors
SL Poynton
Type of isolate
Animal
Year of origin
1991
Cross references
GenBank U94407 Spironucleus vortens alpha-tubulin gene, partial cds.
GenBank U94406 Spironucleus vortens elongation factor-1 alpha gene, partial cds.
GenBank U93086 Spironucleus vortens small subunit rRNA gene, copy 2, partial sequence.
GenBank U93085 Spironucleus vortens small subunit rRNA gene, copy 1 , partial sequence.
GenBank U94408 Spironucleus vortens translation initiation factor-2 gamma subunit gene, partial cds.

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

Poynton S, et al. Spironucleus vortens n. sp. from the freshwater angelfish Pterophyllum scalare: Morphology and culture. J. Eukaryot. Microbiol. 42: 731-742, 1995.

Dawson SC, et al. Stable transformation of an episomal protein-tagging shuttle vector in the piscine diplomonad Spironucleus vortens. BMC Microbiol. 8:71, 2008. PubMed: 18445284.

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