Plasmodium vivax (Grassi and Feletti) Labbe (ATCC® 30073)

Organism: Plasmodium vivax (Grassi and Feletti) Labbe  /  Depositor: National Institutes of Health

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Strain Designations NICA
Application
Emerging infectious disease research
Vector borne research
Biosafety Level 2
Isolation
owl monkey, Nicaragua
Product Format frozen
Type Strain no
Comments
infection in anopheline mosquitoes
transmission from monkey to man
Growth Conditions
Duration: in vivo, Aotus trivirgatus
Protocol: ATCCNO: 30037 SPEC: This strain is distributed as a frozen stabilate. See general instructions for thawing and storage of frozen material before proceeding. As soon as the strain arrives, remove the frozen ampule from the dry ice and transfer it directly to a 35C water bath. When it is completely thawed, aseptically remove the material with a syringe and inject the entire contents of the vial into an Aotus trivirgatus. This parasite can be maintained by serial passage of parasitized blood (by intravenous inoculation) into monkeys (Aotus trivirgatus).
Subcultivation
Protocol: ATCCNO: 30037 SPEC: This strain is distributed as a frozen stabilate. See general instructions for thawing and storage of frozen material before proceeding. As soon as the strain arrives, remove the frozen ampule from the dry ice and transfer it directly to a 35C water bath. When it is completely thawed, aseptically remove the material with a syringe and inject the entire contents of the vial into an Aotus trivirgatus. This parasite can be maintained by serial passage of parasitized blood (by intravenous inoculation) into monkeys (Aotus trivirgatus).
Cryopreservation

CRYOPRESERVATION: 

Only young cells (rings) can be frozen in glycerolyte medium* because their membranes are more robust.

1.     To harvest parasites, inject host with ketamine (0.5–1.0ml depending on host size / weight).

2.     Exsanguinate via the femoral vein using a sterile syringe and Yaeger's anticoagulant**, 1 volume anticoagulant to 4 volumes blood.  Note: Unless there is an exchange transfusion, no more than 10% of estimated total blood volume should be collected over a 2 wk. period.

3.     Centrifuge blood for 5 mins. at 1800 rpm in 50 ml centrifuge tube.

4.     Aspirate supernatant using sterile Pasteur pipet.

5.     Resuspend pellet gently in remaining supernatant.

6.     Slowly add 5 volumes of glycerolyte medium to 3 volumes pellet dropwise via a syringe as follows:

        A.    Add the first volume of glycerolyte and allow the tube to stand for 5 mins. at room temperature.

B.    Add the remaining 4 volumes of glycerolyte and gently agitate.

7.   Aliquot mixture into Nunc screw-capped freezing vials and place in a Nalgene 1C cooling apparatus. Place the apparatus at -80°C overnight and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.).

8.     Plunge vials into liquid nitrogen (-196oC) the next day and store in liquid nitrogen or liquid nitrogen vapor.

       

Name of Depositor National Institutes of Health
Chain of Custody
ATCC <<--National Institutes of Health<<--CDC
References

Coatney GR. The primate malarias. Washington, DC: Government Printing Office; 1971.

Collins WE, et al. Transmission of four Central American strains of Plasmodium vivax from monkey to man. J. Parasitol. 58: 332-335, 1972. PubMed: 4623380

Collins WE, et al. Studies of comparative infectivity of fifteen strains of Plasmodium vivax to laboratory-reared anopheline mosquitoes, with special reference to Anopheles culicifacies. J. Parasitol. 72: 521-524, 1986. PubMed: 3537255

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