CASE CLOSED … what really happened in the 2001 anthrax attacks?

* Under The Mouse Protocol (As Under The Rabbit Protocol), Dr. Ivins Was Tasked With Taking Part In Immunization, Bleeding, Challenge And Observation Of The Animals

Posted by Lew Weinstein on January 1, 2012

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2 Responses to “* Under The Mouse Protocol (As Under The Rabbit Protocol), Dr. Ivins Was Tasked With Taking Part In Immunization, Bleeding, Challenge And Observation Of The Animals”

  1. DXer said

    The protocol, like the passive mouse protocol uploaded at the USAMRIID Electronic Reading Room, details the retro-orbital bleeding to be done. I believe it was the day of challenge and then 30 hours later.

    USAMRIID Research Protocol B01-08 “Passive protection of CBA/J mice….
    http://mrmc.amedd.army.mil/content/foia_reading_room/Protocols/animal%20protocol%20B01-08001%20redacted.pdf

    Here is an example of Dr. Ivins’ early planning of passive studies. I am not trained or experienced in such procedures and so I refer you to the civil deposition testimony for the discussion of animal experiments which I expect to ask Lew to upload in due course.

    http://cryptome.org/ivins/012.pdf

    1) Mice = CBA/J females, 10 per group, about 20 g. Inject intraperitoneally on days -1, 0, 1,1 2 and 3 with one of the following: rabbit anti-rPA antiserum; rabbit anti-AVA antiserum; human anti- AVA IgG; Normal rabbit serum; normal human IgG. On day 0, challenge subcutaneously with 10LD50s of V1B spores. Check mice 3X daily for deaths and note differences in survival as well as time to death differences. If protection is seen, repeat experiment with 10 LD50s of Ames spores. If no protection is seen, drop the challenge dose down to 3-5 LD50s of Vollum 1B spores. Enough animals should be ordered to repeat experiments. Total number of animals = 400

    b) Passive studies in guinea pigs. We can do these studies in the guinea pig animal room in B3. 1) Guinea pigs = Hartley strain, 8 males and 8 females per group, about 350 g at the time
    of the experiment. Inject intraperitoneally on days -1, 0, 1, 2 and 3 with one of the following: rabbit anti-rPA antiserum; rabbit anti-AVA antiserum; human anti-AVA IgG; Normal rabbit serum; normal human IgG. On day 0, challenge intramuscularly with 50 LD50s (5000) of Ames spores. Check guinea pigs 3X daily for deaths and note differences in survival as well as time to death differences. If protection is seen, repeat experiment with 100 LD50s of Ames spores. If no protection is seen, drop the challenge dose down to 10 LD50s of Vollum 1B spores. Enough animals should be ordered to repeat experiments. Total number of animals = 500

    c) Passive studies in rabbits and monkeys (aerosol challenge). and I would be willing to work on this with f it is decided not to contract this out.

    d) For ELISAs and TNAs – mice and guinea pigs – establish serology kinetics with < 10 animals that receive antiserum or IgG. (Contract this out??) For rabbits and monkeys, bleed animals daily. (Contract the tests out??)

    e) Surrogate marker for non-human primate: We can either do this, or contract out the immunizations (and bleeds) …

    c) Assays for immunological correlates – see for comments. d) Characterization of protective antibodies – see for comments.

    Dr. Ivins experience was that bleeding dates needed to be strictly followed.

  2. DXer said

    Is it that Rachel and Ken do not understand what is involved in bleeding an animal to death?

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