Leptomonas barvae (ATCC® PRA-303)

Organism: Leptomonas barvae  /  Depositor: D Maslov

Permits and Restrictions

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Strain Designations 21EC
Biosafety Level 1
Isolation
Intestine of Collaria oleosa (Hemiptera, Miridae), Heredia, Costa Rica, 2003.
Product Format frozen
Type Strain no
Medium ATCC® Medium 1034: Modified PYNFH medium (Available from ATCC as ATCC cat. no. 327-X)
Growth Conditions
Temperature: 25.0°C
Growth condition: grown in the presence of unidentified yeast with 5% heat inactivated fetal bovine serum
Cryopreservation
1.   Harvest cells from a culture that is at or near peak density by centrifugation at ~800 x g for 5 min. Pool the cell pellets into a single tube.

2.   Adjust the concentration of cells to 2.0 x 107/ml.  If the concentration is too low, centrifuge at ~800 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired concentration.

3.   Prepare a 10% (v/v) sterile DMSO solution in fresh medium as follows:  Add the required volume of DMSO to a glass screw-capped test tube and place on ice.  Allow the DMSO to solidify.  Add the required volume of refrigerated medium.  Dissolve the DMSO by inverting several times.  If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.

4.   Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 107 and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution to the start of the freezing process should be no less than 15 min and no longer than 60 min.

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.

7.   The frozen preparations are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8.   To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial enough to cover only the frozen material. Do not agitate the vial.

9.   Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate into 10.0 ml of fresh ATCC medium 1034.

10.          Incubate the tube at 20-25°C with the cap screwed on tightly.

Name of Depositor D Maslov
Year of Origin 2003
References

Maslov DA, et al. Two new species of trypanosomatid parasites isolated from Heteroptera in Costa Rica. J. Eukaryot. Microbiol. 57: 177-188, 2010. PubMed: 20113381

Yurchenko VY, et al. Morphological discordance of the new trypanosomatid species phylogenetically associated with the genus Crithidia. Protist 159: 99-114, 2008. PubMed: 17931968

Yurchenko, VY, et al. Leptomonas costaricensis sp. n. (Kinetoplastea: Trypanosomatidae), a member of the novel phylogenetic group of insect trypanosomatids closely related to the genus Leishmania. Parasitology 133:537-546. Pubmed: 16834819.

Yurchenko, V, et al. An integrated morphological and molecular approach to a new species description in the Trypanosomatidae: the case of leptomonas podlipaevi n. sp., a parasite of Boisea rubrolineata (Hemiptera: Rhopalidae). J Eukaryot Microbiol. 53:103-111.

Cross References

Nucleotide (GenBank) : FJ968532 SSU rRNA gene

Nucleotide (GenBank) : FJ968529 gGAPDH gene

Nucleotide (GenBank) : DQ864310 SL RNA gene

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