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

* With respect to the withdrawal of Ames from RMR-1029 on August 27, 2001 by Dr. George V. Ludwig of the Diagnostic Services Division, GAO should obtain …

Posted by DXer on December 28, 2011


With respect to the withdrawal of Ames from RMR-1029 on August 27, 2001 by Dr. George V. Ludwig of the Diagnostic Services Division,  GAO should obtain …

… and review the protocol titled “Production of Reference and Reagent Antibodies in Goats.”

That research was not sponsored by Bacteriology but instead was sponsored by DSD.

The FBI experts in DSD were the ones known to have made a dried powder from RMR-1029, not Dr. Ivins.

The FBI experts in DSD did not submit a sample of Ames from RMR-1029 that they had used to make a dried powder.




4 Responses to “* With respect to the withdrawal of Ames from RMR-1029 on August 27, 2001 by Dr. George V. Ludwig of the Diagnostic Services Division, GAO should obtain …”

  1. DXer said

    The Army CID investigated missing VEE before 9/11 and recovered vials from a Middle Eastern scientist in New York City who had taken them as mementos of his visit to USAMRIID.

    Was the 2009 investigation looking for VEE that went missing between 1987 and 2008 related? Was it part of the same investigation? What was the result of the 2009 investigation?

    What were the circumstances of the missing VEE? Who was the New York City scientist (reportedly from a MIddle Eastern country) working with at USAMRIID?

    Did the VEE expert at USAMRIID also have virulent Ames from RMR 1029?

    The 302 interview reads:

    “Pre-September 11, 2001, ______ recalls an incident where a foreign scientist, from New York, took a vial of Venezuelan Equine Encephalits,. This scientist took the vial to New York, after being administered the vaccine. ______believes that this scientist was from a Middle Eastern country.
    CID tracked the scientist and recovered the vial. This scientist no longer has access to USAMRIID. ________ advised that the FBI investigated this matter as well.

    Cold comfort
    Originally published April 22, 2009

    The Criminal Investigation Division at Fort Meade has been investigating USAMRIID at Fort Detrick since at least early February. Meade’s CID pursues investigations of serious crimes and sensitive subjects of concern to the Army at regional bases like Detrick, which has no internal investigative arm.
    A News-Post story in February reported that USAMRIID was shutting down most of its bioresearch while it tried to match its inventory to its records, citing an “overage” of BSAT, biological select agents and toxins.

    Meade’s CID, however, isn’t concerned with overstock. Instead, agents are looking for what may have gone missing between 1987 and 2008.

    “It’s possible there are some viral samples missing,” at USAMRIID, Fort Meade public affairs officer Chad Jones confirmed in a phone interview Monday.

    “I don’t know anything else. The investigation is ongoing,” he said.

    The investigation into possible missing pathogens began about the same time Col. John P. Skvorak issued a “stand down” memo halting research operations until an updated inventory is complete. The memo made no mention of missing samples.

    A retired support staff employee who worked in the BSL-4 labs received a visit from Fort Mead’s CID agents in February. Agents wanted to know if he’d taken anything out of the lab between 1987 and 2008, and how easy it was for others to remove samples.

    “I said it was easy enough. It was a lock and key access to the suite of freezers,” the retiree said in an interview.

    In that time period, thousands could’ve accessed the freezers of deadly and/or infectious viral samples, he told investigators. Specifically, the man reported, CID asked about samples of VEE, Venezuelan Equine Encephalitis. According to the Centers for Disease Control, VEE is spread to humans by mosquitoes; symptoms range from mild flu-like illness to brain inflammation, coma and death. Mortality rate is one-third, “making it one of the most deadly mosquito-borne diseases in the United States.”

    Another support staff member in the BSL-3 labs left the job a few years ago. In February, he received a message on his answering machine instructing him to call one of two numbers about missing VEE. The phone numbers connected him to Meade’s CID.

    Perhaps we could find solace in the fact that the Army is trying to impose order on the process after more than 20 years, tracking missing viruses and adding others to its database.

    Aside from the obvious — the possibility that deadly viruses may be floating around out there unsecured — two events, however, preclude us from taking one iota of comfort in this scenario: 1) the construction of a greatly expanded biolabs, which means more germs, more people, more risk; and 2) the government’s own admission that the 2001 anthrax murders were an inside job.

    Asked whether he supports an expansion of biolabs at USAMRIID, the former BSL-3 worker said “No.”

    “Not knowing what I know now. With that many people there, things get sloppy.”

    A belief that inventory controls, stricter protocols and psychological screenings will protect the public from USAMRIID’s dangerous pathogens is na?ve. Even Detrick scientists were reportedly upset at the new controls, according to a Feb. 10 AP story, because they don’t suit USAMRIID’s operations.

    Why? Because germ samples can be easily multiplied in the lab and it’s difficult to track them.

    Now that’s comforting.

  2. DXer said

    The former US Attorney in charge of Amerithrax, in an unrelated case, is in the news today.

    Ex-U.S. Atty. Accuses FBI and Prosecutors of Illegally Obtaining Documents and Lying About It


    Howard contends in his letter to the judge, that after Serrano’s initial meeting with FBI agent Gavin Shea in May 2010, the agent directed him to collect more documents from the company. Howard contends that the since Serrano was then acting as an agent for the FBI, the FBI needed to obtain a search warrant after the initial meeting. No search warrant was obtained.

    The FBI has contended that Serrano came with all the documents and did not collect anything after that first meeting.

    Howard claims in the letter that the government is lying.

    In fact, he writes that Serrano testified during cross examination that he collected documents after his first meeting with the FBI , an admission which “contradicts the government’s repeated representations to the Court and defense counsel that Serrano had obrained all the documents before he approached the FBI.”

    “Further cross-examination (of Serrano) revealed that not only had Serrano taken documents from GDC after meeting with Agent Shea, he actually created false financial records, which the government has introduced at trial, in an effort to curry favor with the government,” Howard wrote.

  3. DXer said

    Judging by his emails and publications, Bruce was not involved in the goat study. Here are his publications from 1998 – 2002. The goat study was done under BSL-3 level. What location? When? Late August 2001? Did it involve a challenge of the animals? If so, did it involve aerosol challenge? Or did it involve a subcutaneous challenge? There is no factual basis to suggest that the 5 ml withdrawn on August 27, 2001 for the DSD study was in any way involved in the anthrax mailings. The GAO nonetheless should obtain and review the goat protocol in understanding the August 27, 2001 withdrawal from RMR 1029 of Ames for use in an experiment by the division that had made a dried powder out RMR-1029 at the request of DARPA.

    1. Ivins BE, Pitt ML, Fellows PF, Farchaus JW, Benner GE, Waag DM, Little SF, Anderson GW Jr, Gibbs PH, Friedlander AM. Comparative efficacy of experimental anthrax vaccine candidates against inhalation anthrax in rhesus macaques. Vaccine 16(11-12):1141-8, 1998.

    Abstract: The authors examined the efficacy of Bacillus anthracis protective antigen (PA) combined with adjuvants as vaccines against an aerosol challenge of virulent anthrax spores in rhesus macaques. Adjuvants tested included i) aluminum hydroxide (Alhydrogel), ii) saponin QS-21 and iii) monophosphoryl lipid A (MPL) in squalene/lecithin/Tween 80 emulsion (SLT). Animals were immunized once with either 50 micrograms of recombinant PA plus adjuvant, or with Anthrax Vaccine Adsorbed (AVA), the licensed human anthrax vaccine. The serological response to PA was measured by enzyme linked immunosorbent assay. Lymphocyte proliferation and serum neutralization of in vitro lethal toxin cytotoxicity were also assayed. In all vaccine groups, anti-PA IgM and IgG titers peaked at 2 weeks and 4-5 weeks postimmunization, respectively. Five weeks postimmunization, animals in all vaccine groups demonstrated PA-specific lymphocyte proliferation and sera that neutralized in vitro cytotoxicity. Six weeks after immunization, the animals were challenged by aerosol with approximately 93 LD50 of virulent anthrax spores. Animals were bled daily for 1 week to monitor bacteremia, and deaths were recorded. Anti-PA ELISA titers in all groups of immunized animals were
    substantially increased 2 weeks after challenge. One dose of each vaccine provided significant protection (> 90%) against inhalation anthrax in the rhesus macaques.

    2. Singh Y, Ivins BE, Leppla SH. Study of immunization against anthrax with the purified recombinant protective antigen of Bacillus anthracis. Infect Immun. 66(7): 3447-8, 1998.

    Abstract: Protective antigen (PA) of anthrax toxin is the major component of human anthrax vaccine. Currently available human vaccines in the United States and Europe consist of alum-precipitated supernatant material from cultures of toxigenic, nonencapsulated strains of Bacillus anthracis. Immunization with these vaccines requires several boosters and occasionally causes local pain and edema. We previously described the biological activity of a nontoxic mutant of PA expressed in Bacillus subtilis. In the present study, we evaluated the efficacy of the purified mutant PA protein alone or in combination with the lethal factor and edema factor components of anthrax toxin to protect against anthrax. Both mutant and native PA preparations elicited high anti-PA titers in Hartley guinea pigs. Mutant PA alone and in combination with lethal factor and edema factor completely protected the guinea pigs from B. anthracis spore challenge. The results suggest that the mutant PA protein may be used to develop an effective recombinant vaccine against anthrax.

    3. 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.

    4. Little SF, Ivins BE. Molecular pathogenesis of Bacillus anthracis infection. Microbes Infect. 1(2):131-9, 1999.

    This review summarizes the current knowledge pertaining to the pathogenesis of infection with Bacillus anthracis relative to the two exotoxins and the capsule. Emphasis is given to the structure and activities of the individual components of
    the exotoxins, their interaction with cells, and the response of macrophages to lethal toxin. Finally, results from vaccination studies are reviewed.

    5. Pitt ML, Little S, Ivins BE, Fellows P, Boles J, Barth J, Hewetson J, Friedlander
    AM. In vitro correlate of immunity in an animal model of inhalational anthrax. J Appl Microbiol. 87(2):304, 1999.

    Abstract: The incidence of anthrax in humans is extremely low. Human vaccine efficacy studies for inhalational anthrax cannot be conducted. The identification of a correlate of protection that predicts vaccine efficacy is crucial for determining the immune status of immunized humans. This surrogate marker of immunity can only be established by using an appropriate animal model. Numerous studies showed that protective antigen (PA) is the principle protective antigen in naturally- or vaccine-induced immunity. However, attempts to correlate the quantity of anti-PA antibodies with protective immunity in the guinea pig model for anthrax and various vaccine formulations have failed. In these studies, we used the licensed anthrax vaccine adsorbed (AVA) in rabbits. Groups of New Zealand white rabbits, 10 or 20 per group, were immunized intramuscularly (two doses, 4 weeks apart) with varying doses of AVA, ranging from a human dose to 1:256 dilution in sterile phosphate-buffered saline (PBS). Control rabbits received PBS/Alhydrogel according to the same schedule. Each rabbit was bled 2 weeks after the second dose, and antibody levels to PA measured by both the quantitative anti-PA IgG ELISA and the toxin-neutralizing antibody (TNA) assay. Rabbits were aerosol-challenged 10 weeks from day 0 with a lethal dose of Ames spores. All the rabbits that received the undiluted and 1:4 dilution of vaccine survived, whereas those receiving the higher dilutions of vaccine (1:16, 1:64 and 1:256) had deaths in their groups. All the controls died. Rabbit survival was compared with the antibody response. Statistical models were used to test for significance of the peak antibody responses to predict survival. Results showed that both the amount of anti-PA IgG and TNA titres present in the sera at the time of the peak antibody response were significant (P < 0.0001) predictors of survival. These results demonstrate that the humoral immune response to AVA can predict protection in the rabbit model of inhalational anthrax.

    6. Hail AS, Rossi CA, Ludwig GV, Ivins BE, Tammariello RF, Henchal EA. Comparison of noninvasive sampling sites for early detection of Bacillus anthracis spores from rhesus monkeys after aerosol exposure. Mil Med. 164(12):833-7, 1999.

    Abstract: Bacillus anthracis, a spore-forming bacterium, is the etiologic agent of anthrax. B. anthracis spores can be aerosolized, are relatively easy to produce, and are capable of producing high mortality when inhaled. The prompt use of postexposure antibiotics combined with vaccination greatly increases the survival rate. Rapid detection of exposure is critical to effective case management. Using common collection swabs, culture medium, and culturing equipment, we compared six different noninvasive sampling sites to determine which might best be used to rapidly detect the presence of B. anthracis spores on rhesus monkeys after aerosolization. The results indicate that the greatest number of spores were deposited in the nares, on the face, and on the haired portions of the head, suggesting that these locations are the most effective sampling sites when attempting to detect B. anthracis aerosol exposure.

    7. Fellows PF, Linscott MK, Ivins BE, Pitt ML, Rossi CA, Gibbs PH, Friedlander AM. Efficacy of a human anthrax vaccine in guinea pigs, rabbits, and rhesus macaques against challenge by Bacillus anthracis isolates of diverse geographical origin. Vaccine 19(23-24):3241-7, 2001. Erratum in: Vaccine 20(3-4):635, 2001.

    Abstract: The efficacy of a licensed human anthrax vaccine (Anthrax Vaccine Adsorbed (AVA)) was tested in guinea pigs, rabbits, and rhesus macaques against spore challenge by Bacillus anthracis isolates of diverse geographical origin. Initially, groups of Hartley guinea pigs were vaccinated at 0 and 4 weeks with AVA, then challenged intramuscularly at 10 weeks with spores from 33 isolates of B. anthracis. Survival among the vaccinated groups varied from 6 to 100%, although there were no differences in mean time to death among the groups. There was no correlation between isolate virulence and variable number tandem repeat category or protective antigen genotype identified. New Zealand white rabbits were then vaccinated with AVA at 0 and 4 weeks, and challenged at 10 weeks by aerosol with spores from six of the isolates that were highly virulent in vaccinated guinea pigs. AVA completely protected the rabbits from four of the isolates, and protected 90% of the animals from the other two isolates. Subsequently, two of these six isolates were then used to challenge rhesus macaques, previously vaccinated with AVA at 0 and 4 weeks, and challenged at 10 weeks by aerosol. AVA protected 80 and 100% of the animals from these two isolates. These studies demonstrated that, although AVA confers variable protection against different B. anthracis isolates in guinea pigs, it is highly protective against these same isolates in both rabbits and rhesus macaques.

    8. Pitt ML, Little SF, Ivins BE, Fellows P, Barth J, Hewetson J, Gibbs P, Dertzbaugh M, Friedlander AM. In vitro correlate of immunity in a rabbit model of inhalational anthrax. Vaccine 19(32):4768-73, 2001.

    Abstract: A serological correlate of vaccine-induced immunity was identified in the rabbit model of inhalational anthrax. Animals were inoculated intramuscularly at 0 and 4 weeks with varying doses of Anthrax Vaccine Adsorbed (AVA) ranging from a human dose to a 1:256 dilution in phosphate-buffered saline (PBS). At 6 and 10 weeks, both the quantitative anti-protective antigen (PA) IgG ELISA and the toxin-neutralizing antibody (TNA) assays were used to measure antibody levels to PA. Rabbits were aerosol-challenged at 10 weeks with a lethal dose (84-133 LD(50)) of Bacillus anthracis spores. All the rabbits that received the undiluted and 1:4 dilution of vaccine survived, whereas those receiving the higher dilutions of vaccine (1:16, 1:64 and 1:256) had deaths in their groups. Results showed that antibody levels to PA at both 6 and 10 weeks were significant (P<0.0001) predictors of survival.

    9. Friedlander AM, Welkos SL, Ivins BE. Anthrax vaccines. Curr Top Microbiol Immunol. 271:33-60, 2002.

    The only impetus for the development of new anthrax vaccines is to protect humans against the intentional use of Bacillus anthracis as a bioterrorist or warfare agent. Live attenuated vaccines against anthrax in domesticated animals were among the very first vaccines developed. This was followed by the development of nonliving component vaccines leading to the eventual licensure of protein-based vaccines for human use in the 1970s. This chapter will review the recent advances in developing protein, live attenuated, and genetic vaccines against anthrax.

    10. Fellows PF, Linscott MK, Little SF, Gibbs P, Ivins BE. Anthrax vaccine efficacy in golden Syrian hamsters. Vaccine 20(9-10):1421-4, 2002.

    Abstract: The efficacy of a licensed human anthrax vaccine (anthrax vaccine adsorbed, AVA) was tested in golden Syrian hamsters against a virulent Bacillus anthracis spore challenge. Groups of golden Syrian hamsters were vaccinated at either 0 and 4 weeks or 0, 4 and 8 weeks, then challenged subcutaneously (s.c.) at 10 weeks with spores of various B. anthracis isolates. Although ELISA and toxin neutralization assays demonstrated high titers, none of the AVA-vaccinated hamsters were protected from challenge or demonstrated a significantly extended time to death compared to that of control animals. The results of the study demonstrate that the golden Syrian hamster is not an appropriate model for investigating human anthrax vaccine efficacy.

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