Leishmania donovani (Laveran and Mesnil) Ross

1. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes. Do not agitate the ampule. Do not leave ampule in water bath after it is thawed.
2. Immediately after thawing, aseptically remove the entire contents of the ampule with a syringe and inject intraperitoneally into an uninfected Golden hamster at least 8 weeks old. Follow the protocol for maintenance in vivo. The course of infection may be longer or shorter than usual depending on percent recovery of the parasite from the frozen state.

1. Inoculate entire infected blood suspension intraperitoneally into the hamster using a 1.0 mL syringe equipped with a 27 gauge 1/2 inch needle. Serial passage after 30-45 days.
2. To infect additional hamsters, harvest the parasites from spleen tissue homogenized in a balanced salt solution (i.e., Tyrodes' Salt solution or similar), approximately 5.0 mL salt solution per spleen. Inoculate 0.3-0.5 mL of spleen homogenate per hamster. Dosage should be a minimum of 2 x 106 amastigotes per hamster. The course of infection will vary with the size of the inoculum.
3. Parasites for inoculation should be collected within 30-45 days of infection. Host hamsters should be at least 8 weeks old.

Add ingredients in the sequence listed. Filter-sterilize.

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