Colpoda inflata (Stokes) Kahl (ATCC® 30917)

Organism: Colpoda inflata (Stokes) Kahl  /  Depositor: DH Lynn

Permits and Restrictions

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Application
testing
Biosafety Level 1
Isolation
dried hay, Toronto, Ontario, Canada, 1970
Product Format frozen
Type Strain no
Comments
Phylogenetic relationships
Medium Medium 802: Sonneborn's Paramecium medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with bacteria
Protocol: ATCCNO: 30916 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in a T-25 tissue culture flask containing 10 ml ATCC medium 802 inoculated 24 hours previously with Enterobacter aerogenes ATCC 13048. Transfer any residual liquid from the vial to the flask. Place at 25C. Excystment should occur within a few days.
Subcultivation
Protocol: ATCCNO: 30916 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in a T-25 tissue culture flask containing 10 ml ATCC medium 802 inoculated 24 hours previously with Enterobacter aerogenes ATCC 13048. Transfer any residual liquid from the vial to the flask. Place at 25C. Excystment should occur within a few days.
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 DH Lynn
Year of Origin 1970
References

Greenwood SJ, et al. Phylogenetic relationships of Blepharisma americanum and Colpoda inflata within the phylum ciliophora inferred from complete small subunit rRNA gene sequences. J. Protozool. 38: 1-6, 1991. PubMed: 1900085

Wickham SA, Lynn DH. Relations between growth rate, cell size, and DNA content in Colpodean Ciliates (Ciliophora: Colpodea). Eur. J. Protistol. 25: 345-352, 1990.

Bowers NJ, Pratt JR. Estimation of genetic variation among soil isolates of colpoda inflata (Stokes) (Protozoa: Ciliophora) using the polymerase chain restriction fragment length polymorphism analysis. Arch. Protistenkd. 145: 29-36, 1995.

8310 D: Growth inhibition test with the soil ciliate Colpoda inflata. Washington, DC:American Public Health Association;Standard Methods for the Examination of Water and Wastewater, 2005 APHA8310 - Ciliated Protozoa;.

Cross References

Nucleotide (GenBank) : M97908 Colpoda inflata small subunit ribosomal RNA gene sequence.

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