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Dileptus americanus Kahl

50259

Product category
Protists
Classification
KINGDOM: Protozoa
Strain designation
RV
Type strain
No
Isolation source
Temporary pool of water in hickory tree stump
Geographical isolation
United States; Maryland; Rockville
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.

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.

Raoultella planticola (Bagley et al.) Drancourt et al.

700831

Price: $376.00 ea

Klebsiella aerogenes Tindall et al.

13048

Price: $72.00 ea

Detailed product information

Handling information

Medium
Instruction for complete medium
ATCC® Medium 802 inoculated with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™). ATCC Medium 802 will support growth of b
Temperature
20-25°C
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 ampoules 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 such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after it is thawed.  Do not allow ampule to overheat.
  2. Add the thawed contents to a T-25 flask containing 10 mL of ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).
  3. Aseptically transfer 1-2 mL of washed Tetrahymena to the T-25 flask (see section on CULTURE MAINTENANCE below).  Incubate the culture at 20-25°C.
Handling notes
This strain must be fed with live Tetrahymena (i.e., ATCC® 30005™ or similar, not provided).  The Tetrahymena should be maintained separately and fed to Dileptus at regular intervals.  Dileptus will eventually form cysts once the supply of prey organism has been depleted.
Culture maintenance

Periodically add prey organisms as follows:

  1. Maintain growing cultures of Tetrahymena separately at 20-25°C in T-25 tissue culture flasks containing 10 mL ATCC medium 1034 without serum.
  2. Prepare washed Tetrahymena as follows:  Remove 5-10 mL from a culture at or near peak density, centrifuge at 300 x g for 3 min, quickly remove most of the supernatant (leaving approx. 1 mL), then resuspend cells in 10 mL ATCC medium 1323 or similar.  Centrifuge and resuspend cells again as above.  Repeat this washing step at least twice.
  3. When the Dileptus have consumed all prey Tetrahymena, add 0.5-1.0 mL of washed Tetrahymena prepared in step 2.  The feeding interval will depend on the number of predators present and the culture density of the washed prey.
  4. The Dileptus may be passaged to a T-25 flask by gently rubbing the flask surface with a spread bar to dislodge any attached cells or cysts, then transferring 0.5 to 2 mL to a fresh T-25 flask containing 10 mL ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).  Incubate the culture at 20-25°C, feeding periodically with washed Tetrahymena as described above.
Reagents for cryopreservation
Cryoprotective Solution
DMSO, 1.5 mL
Fresh growth medium w/o bacteria, 8.5 mL
Cryopreservation

Harvest and Preservation
  1. Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.
  2. Harvest Dileptus cells or cysts from a culture that has recently passed peak density by filtration and centrifugation at 200-300 x g for 5 min.
  3. Adjust the concentration of cells or cysts at least 2 x 104/mL in fresh medium.
  4. Mix the cell preparation and the cryoprotective solution in equal portions.
  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 vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through heat of fusion. At -40°C plunge ampules 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. Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
  8. To establish a culture from the frozen state place the vial in a 35°C water bath. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial. Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and inoculate into a T-25 tissue culture flask containing 10 mL of ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).
  9. Aseptically transfer 1-2 mL of washed Tetrahymena to the T-25 flask (see section on CULTURE MAINTENANCE).  Incubate the culture at 20-25°C.
    Once the culture is established, follow the protocol for maintenance of culture.

History

Deposited as
Dileptus americanus Kahl
Depositors
EB Small
Chain of custody
ATCC <-- EB Small <-- T.A. Nerad
Type of isolate
Environmental
Year of origin
1986

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.

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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.

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References

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