* Under the animal protocols, there would be 3X’s checks (including a night check) and daily check for bacteremia and determination of serum antibody levels; for a subcutaneous challenge done in Building 1425, Dr. Ivins would do the bacteremia study in Rm. 313.
Posted by DXer on November 8, 2011
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CASE CLOSED ... what really happened in the 2001 anthrax attacks? 
DXer said
In this email, Dr. Ivins is asking how many times the animals would be checked.
But even when animals are checked only twice, what do protocols or experts say about the preferred timing of the checks? Is it specified?
More humane care in experiments involving terminal endpoints would be 3Xs rather than 2Xs — and where it is 2Xs, it would be scheduled in such a way as to have the least amount of time between checks — so as to avoid prolonged suffering. Thus, for example, a check at 8 a.m. and 4 p.m. would leave 16 hours between the two checks. Very bad IMO. A check at 8 a.m. and 8 p.m. would leave a window of 12 hours. Better but still not good. What do the experts say about the optimum scheduling of animal checks in animal experiments in which the animals are killed by anthrax over a period of days.
Of course, the protocol necessarily would require that animal checks be made daily — which would include weekends.
For Agent Lawrence Alexander and Dr. Vahid Majidi and Rachel Lieber to claim that Ivins had no reason to come in the weeks is totally false and, without more, dispositive of their cotton candy “Ivins Theory.”
He necessarily was OBLIGATED to be coming in on the weekend after the challenge. It was mandated by the animal protocol. Their claim was contradicted by the documents that were available to them upon any sort of diligent review of the contemporaneous documents.
Dr. Ivins had tried to get back the relevant documents from USAMRIID (emails) and FBI (Lab Notebook 4241) but was refused them. (I’ve uploaded his correspondence and other documents on his inability to get the documents).
The prosecutors and investigators were operating in what Hillary Clinton would call an “evidence free” zone. They were substituting their suppositions and conclusory assertions for the contemporaneous documents — contenting themselves with the fact that Dr. Ivins could not remember or reconstruct his time in detail without the documents.
In citing his interrogation statement recalling problems at home — and the office being right near his home — she made a silk purse out of a sow’s ear.
What were YOU doing the first week of October seven years ago? Huh? What did you say? You can’t remember? You must be guilty of murder.
Problems at home you say? We are going to call your wife and children before the grand jury and ask them about the stained panties we found in your basement.
Amerithrax represents the greatest failure in counterintelligence analysis in the history of United States.
http://www.amerithrax.wordpress.com
DXer said
Note that in the experiment above involving an aerosol challenge to monkeys, that would have to be conducted in Building 1412 / aerobiology rather than Building 1425 — and thus night checks would be logged there.
By providing statistics related to Building 1425, and not understanding that the two-person rule was implemented in January 2002, the FBI’s data, intentionally or not, was totally misleading.
To be meaningful, a powerpoint presentation by Agent Alexander to AUSA Lieber would have required an analysis of Dr. Ivins hours in both buildings and the substantive reasons for changes.
DXer said
As Old Atlantic, I believe, noted, this January 2000 email does not provide authority for the headline I had written. It merely had a question mark after “3”. Moreover, in any event, the number of daily animal checks would depend on the actual approved protocol for the specific experiment.
DXer said
The greatest number of rabbits would have been dying the day before the start of the second window of mailing. The investigator and prosecutor evidences no awareness whatsoever of the October 2, 2001 challenge of rabbits or the October 5, 2001 email about the number of rabbits that had died in the first 3 days after challenge.
Zaucha GM, Pitt LM, Estep J, Ivins BE, Friedlander AM. The pathology of experimental anthrax in rabbits exposed by inhalation and subcutaneous inoculation. Arch Pathol Lab Med.122(11):982-92, 1998.
Abstract: OBJECTIVE: Although rhesus monkeys are considered to be an appropriate model for inhalational anthrax in humans, an alternative for vaccine and therapeutic efficacy studies is desirable. This study characterized the pathology of lethal anthrax in rabbits challenged by subcutaneous inoculation and aerosol exposure. MATERIALS AND METHODS: New Zealand white rabbits were exposed by subcutaneous inoculation or aerosol to lethal doses of Bacillus anthracis spores. RESULTS: The pathology of anthrax in rabbits exposed by either route was similar, with principal findings occurring in the spleen, lymph nodes, lungs, gastrointestinal tract, and adrenal glands. The cardinal changes were hemorrhage, edema, and necrosis, with bacilli and limited leukocytic infiltration. Features that depended on the route of exposure included mediastinitis in aerosol-exposed rabbits, a primary dermal lesion after subcutaneous inoculation, and differences in the pattern of lymph node involvement. Lesions observed in rabbits were comparable to those of inhalational anthrax in humans and rhesus monkeys. Noteworthy differences included the lack of leukocytic infiltration in brain and meningeal lesions, the relatively mild mediastinal lesions, and a lower incidence of anthrax-related pneumonia in rabbits compared with humans. These differences may be attributed to the greater susceptibility of rabbits to anthrax. Increased susceptibility is associated with both reduced leukocytic response to the bacilli and a more rapid progression to death, which further limits development of leukocytic infiltrates in response to the basic lesions of hemorrhage and necrosis. Primary pneumonic foci of inhalational anthrax, which may be influenced by preexisting pulmonary lesions in humans, were not observed in our rabbits, which were free of preexisting pulmonary disease. CONCLUSION: Anthrax in rabbits may provide a useful model for evaluating prophylaxis and therapy against inhalational anthrax in humans.
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For subcutaneous inoculation, spores were suspended in sterile water for injection and then heat shocked at 60 deg C for 45 minutes. Appropriate dilutions were prepared, to achieve the desired dose of spores in a final volume of 0.5 mL/ dose. Spore dilutions were held on ice until administered. Actual spore counts in the inoculum were verified by quantitative bacterial culture. Rabbits were inoculated with 0.5 mL of the material (dose range, 43 to 1.56 x 10^sup 5^ colony-forming units (CFU); subcutaneous LD^sub 50^ = 1.56 x 10^sup 3^ CFU; subcutaneous lethal dose^sub 99^ (LD^sub 99^) = 2.83 x 10^sup 6^ CFU) in the dorsal interscapular region.
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Rabbits included in the pathology study died 2 to 4 days after exposure to B. anthracis spores, with mean survival times of 2.9 days and 2.4 days for subcutaneously inoculated and aerosol-exposed rabbits, respectively. Although there was a trend for decreased survival time with increasing dose, the effect was minimal (Table 1). Fulminant disease appeared to be an all-or-none response, and no protracted illness was observed, regardless of the dose. Clinical signs were not generally apparent until within 24 hours of death, at which time rabbits became progressively lethargic and weak. Several rabbits, later found to have brain or meningeal lesions, exhibited brief periods of excitation and hyperactivity within hours or minutes before death.
DXer said
In previous years, as illustrated by the study above, checks commonly were only done 2Xs. Protocols came to be modified to include checks 3Xs which led to increased night hours in the B3 in Building 1425 in the case of subcutaneous challenges. By April 2002, in addition to the new regulations imposed, Dr. Ivins work in the suite was curtailed after the brouhaha about the contamination. see his emal to this effect. Thus, not only is the argument about hours not valid when apples and apples are compared (other 2001 subcutaneous challenges) but it is specious for these bigger picture reasons over an extended period.
Rabbits would continue to be monitored for up to two weeks. Bacteremia in all of these rabbit studies was always closely studied.
See also
Efficacy of a human anthrax vaccine in guinea pigs, rabbits, and rhesus macaques against challenge by Bacillus anthracis isolates of diverse geographical origin
P. F. Fellows a,*, M. K. Linscott a, B. E. Ivins a, M. L. M. Pitt b, C. A. Rossi c, P. H. Gibbs d, A. M. Friedlander a
a Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA b Toxinology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA
c Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA,
d Biometrics and Information Management Division4, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, 21702-5011, USA
Received 5 September 2000; received in revised form 11 December 2000; accepted 18 December 2000
Vaccination and challenge schedule
Hartley guinea pigs, New Zealand white rabbits and rhesus macaques were vaccinated at 0 and 4 weeks i.m. with 0.5 ml of AVA. All control animals received 0.5 ml of saline. Challenge occurred 10 weeks after the first vaccination and animals were monitored for survival for 14 days. Death by anthrax was confirmed in all animals by plating blood on tryptic soy agar (TSA) and incubating overnight at 37oC.
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In contrast to guinea pigs, rabbits were protected by vaccination with AVA. This is significant in that the immunized rabbits survived aerosol challenge doses of spores that were equivalent to 300-2700 aerosol LD50s of the Ames isolate. Although the animals were protected from death at these high challenge doses, several demonstrated a transient bacteremia. This is noteworthy in that previous studies in which rabbits were vaccinated with AVA and challenged with the Ames isolate, no bacteremia was found in any of the rabbits [29]. However, because LD50 determinations were not performed for the isolates in these studies, it is unknown whether the reported bacteremias were the result of high challenge doses or of other unidentified factors.
oldatlantic said
Where is the support for these statements? They are anticipations of the documents not yet released?
DXer said
Below is where I uploaded an excerpt from a representative rabbit protocol. Over the past days, I’ve “cc’ed” to your inbox various FOIA requests directed to the wonderful FOIA person who handles requests relating to USAMRIID.
I believe the computer document we are looking for is titled something like ADDENDUMrabbitprotocol
oldatlantic said
I appreciate your hard work.
DXer said
Dr. Ivins appears to have written the document that he titled ADDENDUMrabbitprotocol by May 24, 2001. He transmitted the “B0-03 addendum” by email dated May 24, 2001.
There thus are three documents titled ADDENDUMrabbitprotocol, ADDENDUMrabbitprotocol02, and ADDENDUMrabbitprotocol03 that are of interest and likely differ in minor respects.
DXer said
Dr. Ivins sent a corrected electronic copy of his animal protocol by email dated July 6, 2001. USAMRIID is willing to provide a copy of attachments to emails but it helps if specific attachments of interest are identified — given that there are nearly a decade’s worth of emails.
DXer said
The committee that reviews the use of animals provided their response transmitted by email July 6, 2001. I will request a copy from USAMRC under FOIA.
From:
Sent: Friday, July 06, 2001 9:12 AM
To: Ivins, Bruce E Dr USAMRIID
Subject: LACUC Review of Proposal 01-28
Bruce, See attached minutes.
File: 21Jun01.bi.doc >>
DXer said
An email dated August 9, 2001 from someone attaches modifications to his rabbit protocol to address protection by the pure isoforms. The file is named File: Isoform Rabbit protocol.doc. (The isoform study was separate from the study about whether they could stop using formaldehyde; formaldehyde was discouraged by the FDA.)