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Stephanopogon apogon Borror

50096

Product category
Protists
Strain designation
BCP
Type strain
No
Isolation source
Seawater from Cliff Pool
Geographical isolation
Bemuda
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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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

Characteristics

Comments
Food source, Rhynchomonas nasuta ATCC 50105, not supplied. Mixed bacteria present.
Ultrastructural identity

Handling information

Medium
Instruction for complete medium

Media: ATCC® Medium 1525 and one of the alternate media listed below, combined in equal parts and inoculated with Klebsiella pneumoniae subsp. pneumoniae (ATCC 700831™) or Enterobacter aerogenes (ATCC 13048™)

Alternate media: ATCC® Medium 1405 HESNW medium, ATCC® Medium 1361 Marine Flagellate medium (both media augmented with two sterile rice grains added to culture flask)

Note about growth media: ATCC Media 1405 and 1361 will each support moderate growth of the prey organism when used on their own, but will support more rapid and dense growth when combined in an equal-parts mixture with previously-bacterized ATCC Medium 1525.

Temperature
25°C
Culture system
Xenic
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 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 an equal-parts mixture of ATCC medium 1525 and either ATCC medium 1405 or ATCC medium 1361, bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC 700831™) or Enterobacter aerogenes (ATCC 13048™).
  3. Aseptically transfer 1-2 ml from a thriving culture of Rhynchomonas to the T-25 flask. Incubate the culture at 25°C with the cap screwed on tightly.
Culture maintenance
Subculture every 7-14 days to a fresh T-25 flask of the prey organism in bacterized medium as follows:
  1. Maintain growing cultures of Rhynchomonas separately in T-25 tissue culture flasks containing 10 ml of an equal-parts mixture of ATCC medium 1525 and either ATCC medium 1405 or ATCC medium 1361, bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC 13048).
  2. When the prey organism is at or near peak density, rub the bottom surface of an encysted culture of Stephanopogon with a cell scraper to detach the cysts, vigorously agitate, and aseptically transfer a 0.5 ml aliquot of the cyst suspension to the prey culture.  Incubate at 25°C with the cap screwed on tightly.
Reagents for cryopreservation

Cryoprotective Solution
DMSO, 2.0 ml
Fresh growth medium w/o bacteria, 8.0 ml

Cryopreservation
  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 Stephanopogon cells or cysts from a culture that has recently passed peak density by filtration and centrifugation at 800 x g for 5 min.
  3. Adjust the concentration of cysts at least 2 x 105/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 an equal-parts mixture of ATCC medium 1525 and either ATCC medium 1405 or ATCC medium 1361, bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC 700831?) or Enterobacter aerogenes (ATCC 13048™).
  9. Aseptically transfer 1-2 ml from a thriving culture of Rhynchomonas to the T-25 flask. Incubate the culture at 25°C with the cap screwed on tightly.

Once the culture is established, follow the protocol for maintenance of culture.

History

Deposited as
Stephanopogon apogon Borror
Depositors
EB Small
Type of isolate
Environmental
Year of origin
1984

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

Patterson DJ, Brugerolle G. The ultrastructural identity of Stephanopogon apogon and the relatedness of the genus to other kinds of protists. Eur. J. Protistol. 23: 279-290, 1988.

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