Cercomonas vibrans Sandon (ATCC® 50530)

Organism: Cercomonas vibrans Sandon  /  Depositor: J Martinez-Cruz

Permits and Restrictions

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Deposited As Cercobodo sp.
Biosafety Level 1
Isolation
dry soil, Mexico City, Mexico
Product Format test tube
Type Strain no
Comments
This is the only species of Cercomonas known to feed on other protists.
Medium ATCC® Medium 1967: Soil freshwater cereal grass medium
Growth Conditions
Temperature: 25.0°C
Protocol: This strain is shipped as a growing culture and includes Adriamonas sp. (food source for Cercomonas vibrans) and Enterobacter aerogenes ATCC 13048 (food source for Adriamonas sp.). The food will not be completely eliminated by the predator, C. vibrans. The Adriamonas sp. will eventually encyst. C. vibrans also produces cysts. Because those cysts are not very stable, the culture must be passaged every 7-10 days to ensure the culture's continued viability. Upon arrival of the shipment, agitate the culture, aseptically transfer the entire contents to a T-25 tissue culture flask, and incubate it at 25C. The bacterial flora present in the shipped culture will support growth of the Adriamonas sp., but growth can be improved by using medium that has been bacterized with E. aerogenes ATCC 13048 approximately 24 hours before inoculation with the Cercomonas/Adriamonas culture. (Other species of bacteria may work equally well, but this one has been empirically assessed.) After bacterization of the medium, agitate the culture of ATCC 50530 and aseptically transfer 0.25 ml to the new medium. Screw the cap on tightly and incubate at 25C.
Subcultivation
Protocol: This strain is shipped as a growing culture and includes Adriamonas sp. (food source for Cercomonas vibrans) and Enterobacter aerogenes ATCC 13048 (food source for Adriamonas sp.). The food will not be completely eliminated by the predator, C. vibrans. The Adriamonas sp. will eventually encyst. C. vibrans also produces cysts. Because those cysts are not very stable, the culture must be passaged every 7-10 days to ensure the culture's continued viability. Upon arrival of the shipment, agitate the culture, aseptically transfer the entire contents to a T-25 tissue culture flask, and incubate it at 25C. The bacterial flora present in the shipped culture will support growth of the Adriamonas sp., but growth can be improved by using medium that has been bacterized with E. aerogenes ATCC 13048 approximately 24 hours before inoculation with the Cercomonas/Adriamonas culture. (Other species of bacteria may work equally well, but this one has been empirically assessed.) After bacterization of the medium, agitate the culture of ATCC 50530 and aseptically transfer 0.25 ml to the new medium. Screw the cap on tightly and incubate at 25C.
Cryopreservation
Cryoprotective Solution

DMSO                                                                                    2.0 ml

Fresh growth medium w/o bacteria                                 8.0 ml

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 cells from a culture that is at or near peak density by filtration and centrifugation at 800 x g for 5 min.

3.  Adjust the concentration of cells at least 2 x 106/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 ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC® 13048).

9.     Incubate at 25°C with the cap screwed on tightly.

10.   Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 ml to 10.0 ml of bacterized ATCC medium 802.

11.   Follow the protocol for maintenance of culture.

Name of Depositor J Martinez-Cruz
Chain of Custody
ATCC <<--J Martinez-Cruz<<--T.A. Nerad
References

Jovita Martinez-Cruz, personal communication

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.
Basic Documentation