Crithidia oncopelti (Noguchi and Tilden) Hanson and McGhee (ATCC® 30264)

Depositor: KP Chang  /  Biosafety Level: 1

Permits and Restrictions

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Biosafety Level 1
Isolation
Aposymbiotic
Product Format frozen
Type Strain no
Comments
species description
Ornithine-arginine metabolism
Ultrastructure of symbiotic bacteria
Proteolytic activities
isoleucine requirement and threonine deaminase
endonuclease-generated fragments of K-DNA, esterase isoenzymes, surface proteins for species identification
Acetylornithinase and ornithine acetyltransferase
Ultrastructural differences between species with and without endosymbionts
Medium Medium 355: Crithidia medium
Growth Conditions
Temperature: 25.0°C
Duration: axenic
Protocol: ATCCNO: 11745 SPEC: See general instructions for thawing and storage of frozen material before proceeding. Add thawed contents to a single 16 x 125 mm glass screw-capped test tube of the appropriate medium. Incubate the culture vertically with the cap screwed on tightly. It is essential to establish cultures initially in small volumes. Once established, the culture can be scaled up to larger volumes. Vigorously agitate the culture and aseptically transfer 0.1 ml of culture to a fresh tube of medium weekly.
Subcultivation
Protocol: ATCCNO: 11745 SPEC: See general instructions for thawing and storage of frozen material before proceeding. Add thawed contents to a single 16 x 125 mm glass screw-capped test tube of the appropriate medium. Incubate the culture vertically with the cap screwed on tightly. It is essential to establish cultures initially in small volumes. Once established, the culture can be scaled up to larger volumes. Vigorously agitate the culture and aseptically transfer 0.1 ml of culture to a fresh tube of medium weekly.
Cryopreservation

1.   Prepare a 10% (v/v) sterile DMSO solution in fresh ATCC Medium 1034. 

2.   Transfer a culture at peak density to centrifuge tubes and centrifuge at 525 x g for 5 minutes.

3.   Remove the supernatant and resuspend the cells in ATCC medium 1034 to a concentration of 2 x 106 to 2 x 107 cells/ml.

4.   Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/ml and 5% (v/v) DMSO.

5.   Distribute the cell suspension in 0.5 ml aliquots into 1.0-2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).  The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 30 min.

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 are stored in either the vapor or liquid phase of a nitrogen freezer.

8.   To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial.

9.   Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5 ml of fresh ATCC medium 1034 in a 16 x 125 mm screw-capped test tube. Incubate upright at 25°C with caps screwed on tightly.

Name of Depositor KP Chang
Chain of Custody
ATCC <<--KP Chang<<--ATCC 12982
References

Chang KP. Ultrastructure of symbiotic bacteria in normal and antibiotic-treated Blastocrithidia culicis and Crithidia oncopelti. J. Protozool. 21: 699-707, 1974. PubMed: 4217371

Figueiredo EN, et al. Enzymes of the ornithine-arginine metabolism of trypanosomatids of the genus Crithidia. J. Protozool. 25: 546-549, 1978.

Chang KP, et al. Effects of methylglyoxal bis(ganylhydrazone) on trypanosomatid flagellates: inhibition of growth and nucleoside incorporation in Trypanosoma brucei. J. Protozool. 25: 145-149, 1978. PubMed: 660567

Faria e Silva PM, et al. Herpetomonas roitmani (Fiorini et al., 1989) n. comb.: a trypanosomatid with a bacterium-like endosymbiont in the cytoplasm. J. Protozool. 38: 489-494, 1991. PubMed: 1920148

Camargo EP, et al. Proteolytic activities in cell extracts of trypanosomatids. J. Parasitol. 64: 1120-1121, 1978. PubMed: 739304

Galinari S, Camargo EP. Trypanosomatid protozoa: survey of acetylornithinase and ornithine acetyltransferase. Exp. Parasitol. 46: 277-282, 1978. PubMed: 569594

Freymuller E, Camargo EP. Ultrastructural differences between species of trypanosomatids with and without endosymbionts. J. Protozool. 28: 175-182, 1981. PubMed: 7024533

Alfieri SC, Camargo EP. Trypanosomatidae: isoleucine requirement and threonine deaminase in species with and without endosymbionts. Exp. Parasitol. 53: 371-380, 1982. PubMed: 6806116

Camargo EP, et al. Electrophoretic analysis of endonuclease-generated fragments of k-DNA, of esterase isoenzymes, and of surface proteins as aids for species identification of insect trypanosomatids. J. Protozool. 29: 251-258, 1982. PubMed: 6284925

Cross References

Nucleotide (GenBank) : U67436 Crithidia oncopelti large subunit 24S rRNA gene, 5'end, LS1 region.

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