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

* “More bad irradiation news” – USAMRIID’s Bruce Ivins found that sometimes samples that had tested negative then upon retesting came up “hot”

Posted by Lew Weinstein on June 19, 2015

“More bad irradiation news” – USAMRIID’s Bruce Ivins found that sometimes samples that had tested negative then upon retesting came up “hot”

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16 Responses to “* “More bad irradiation news” – USAMRIID’s Bruce Ivins found that sometimes samples that had tested negative then upon retesting came up “hot””

  1. DXer said

    While USAMRIID delays further in producing the requested documents relating to shipments involving Dugway and USAMRIID, let’s touch some highlights.

    Dugway, for example, shipped 340 ml Ames spores for irradiation on Jun 27, 01 and 175 ml Ames spores for irradiation on August 30, 2000.

    https://vault.fbi.gov/Amerithrax/Amerithrax%20Part%2036%20of%2059

    A 2004 FBI report indicates that after irradiation of spores sent from Dugway, spores were kept in warehouse cold room near loading dock – even though Ivins group once found irradiation had not been successful in killing the spores
    .
    https://vault.fbi.gov/Amerithrax/Amerithrax%20Part%2036%20of%2059

    It is odd that the FBI officials leading the DOD Review did not detect any problems even though they were on notice of the problems.

    The FBI has not disclosed the Microsoft Excel spreadsheets summarizing pertinent dates from Notebook regarding RMR 1029 (containing Dugway Ames) and a log detailing dates that B. anthraces Ames was irradiated.

    FBI memo, dated May 21, 2004
    https://vault.fbi.gov/Amerithrax/Amerithrax%20Part%2036%20of%2059

  2. DXer said

    Congressman Collins of New York had very interesting substantive remarks about best practices in private industry relating to anthrax.

    He says a spore can pop up live 5 months later — that 48 hours on culture is wholly insufficient. I would have to defer to an expert on that.

    I only point out that Bruce Ivins provided 40 ml of irradiated Ames to be rendered into a powder to the FBI’s anthrax expert, John Ezzell, who then provided the irradiated (then powderized) Ames to DARPA researchers.

    Bruce Ivins would wait 48 hours, I believe — and I think the Ames anthrax in 2000 at USAMRIID was irradiated at 4 million rads.

    Then those samples provided to the FBI’s expert (Dr. Ezzell) were destroyed and no documents were kept. See FOIA response.

    The Ames went to places like Johns-Hopkins and University of Maryland.

    I would further point that Dr. Ivins found that sometimes irradiated Ames would first test negative and then test positive — prove to be live. See email above.

    With this as background, although I found Dr. Christian Hassell to be well-spoken and to inspire confidence, he is avoiding the key lesson to be learned. He was the head of the FBI Laboratory at the time of Amerithrax.

    Thus, who is responsible for setting things right? He is. Who is accountable? The guy who finesses the issue and avoids learning the critical lesson that needs to be taken to heart.

    Both Vahid Majidi and Chris Hassell will be the fellows to fire if and when Dr. Ayman Zawahiri attack the US in a mass anthrax attack.

    Anthrax, Al Qaeda and Ayman Zawhiri: The Infiltration Of US Biodefense
    http://www.amerithrax.wordpress.com

    • DXer said

      I also found the CDC representative articulate but for him not to acknowledge that the strain was the Ames strain — the same strain as in the 2001 attacks — and instead spin the response to focus on whether it was in liquid form or powder — was really wrong.

      He even used the phrase “natural strain”. Ames, of course, was a natural strain — originating from a cow in Texas many years ago.

      Once an individual has access to virulent Ames, he can then grow more, concentrate it, and then dry it.

  3. DXer said

    “DPG [Dugway] personnel noted in the retesting experiments currently underway, multiple Bacillus species including B. anthracis strains that were inactivated and archived have come up positive (i.e., viable spores are present in inactivated samples).”

    “Review Committee Report: Inadvertent Shipment of Live Bacillus anthraces spores by DoD,” July 13, 2015, p. 34.

  4. DXer said

    Compare

    “Why post inactivation viability testing did not detect the presence of live BA:

    There is no single root cause to explain why the BA samples were incompletely inactivated, or why viability testing did not detect live BA spores. Contributing factors that may have resulted in undetected live BA spores during viability testing in DPG’s samples include deficiencies in sample sizes and inadequate incubation periods after irradiation. ”

    “Review Committee Report: Inadvertent Shipment of Live Bacillus anthraces spores by DoD, July 13, 2015

  5. DXer said

    http://www.koreatimes.co.kr/www/news/nation/2015/06/116_181360.html

    Complaint filed against two USFK commanders over anthrax delivery

    By Lee Kyung-min

    A group of activists filed a complaint with the prosecution Monday against two U.S. Forces Korea (USFK) commanders for their alleged involvement in the delivery of live anthrax samples to a military lab here from the United States.

    During a press conference near the Seoul Central District Prosecutors’ Office, the activists said the two ― USFK Commander Gen. Curtis Scaparrotti and 7th Air Force Commander Lt. Gen. Terrence O’Shaughnessy ― should be held accountable for the unlawful delivery of the hazardous material.

    Earlier, the USFK admitted that live anthrax samples were delivered to its Osan Air Base, south of Seoul, for tests, triggering uproar here. The Pentagon apologized for the delivery, but said it was an “accident.”

    “The USFK failed to comply with the law here, thereby posing substantial public health risk. We demand a thorough investigation into the matter and due punishment for the two,” the activists said in a statement.

    Comment:

    There was not a substantial public health risk IMO.

    Moreover, the commanders are in no way responsible, IMO.

    Now if there were inadequate radiation of other materials that are infectious and transmitted persom-to-person that would be more noteworthy and of greater concern. Was there? Are other agents being tested? If not, why not?

  6. DXer said

    Bruce Ivins sent out irradiated Ames. The fact that it sometimes would test positive and grow — after testing negative — further undermines the already demolished reasoning relied upon by the FBI relating to the distribution of virulent Ames.

    But a public confession would help keep things simpl in a matter that the FBI made far more complex than it needed to be. When does the trial of my former Facebook Friend Yazid Sufaat resume? In written correspondence, he does not deny responsibility for the Fall 2001 anthrax mailings.

    In written correspondence with the Case Closed blog, the Sacramento State graduate and former Malaysian Army Captain Yazid Sufaat has invoked the “Fifth Amendment” and declined to disclose the b. anthracis strain he was using.
    Posted by Lew Weinstein on April 28, 2015
    https://caseclosedbylewweinstein.wordpress.com/2015/04/28/in-written-correspondence-with-the-case-closed-blog-the-sacramento-state-graduate-and-former-malaysian-army-captain-yazid-sufaat-has-invoked-the-fifth-amendment-and-declined-to-disclose-the-b/

    If Yazid Sufaat and his assistants had not been working with virulent anthrax at the Kandahar lab, there would have been no reason for them to have been vaccinated
    Posted by Lew Weinstein on September 22, 2014
    https://caseclosedbylewweinstein.wordpress.com/2014/09/22/if-yazid-sufaat-and-his-assistants-had-not-been-working-with-virulent-anthrax-at-the-kandahar-lab-there-would-have-been-no-reason-for-them-to-have-been-vaccinated/

    Anthrax, Al Qaeda and Ayman Zawahiri: The Infiltration of US Biodefense
    http://www.amerithrax.wordpress.com

    Off-topic:

    To New York State Governor Cuomo’s office:

    The Governor’s father once said:

    “Every time I’ve done something that doesn’t feel right, it’s ended up not being right.”

    Now that the Assembly passed the bill relating to mute swans 132-15 (after the Senate passed the bill 60-1), the Governor should take care in deciding whether to veto the bill in an unthinking political-minded deference to the hunter’s lobby.

    The bill merely requires that the DEC decision-making be science-based and that the DEC demonstrate the claimed environmental harm caused by mute swans. The bill requires two hearings be held. When did democracy become a dirty word?

    When did science-based reasoning (as distinguished from speculative claims) become objectionable?

    Mute swans are a sentinel species and warn of continuing contamination. The Governor this year is spending $50 million here to build an amphitheater on uncharacterized industrial wastebeds on the Onondaga Lake shoreline. If the Governor vetoes the mute swan bill and his employees shoot my daughter’s mute swans, then know that I’ll be looking hard to prove continuing mercury and other very dire contamination. Both the filmed shooting of swans and endangering the health of New Yorkers will end up being the Governor’s legacy. I have experience in testing water and so it will be easy to do.

    In contrast, the mute swans would make a lovely landscaping for his amphitheater. So leave my daughter’s swans alone and we’ll both be able to enjoy the new amphitheater with a clear conscience. When you are next in town, go see my exhibit at the Everson museum for plans for an eagle and mute swan viewing station.

    I’ve worked hard to keep New York City safe from a mass anthrax attack by Dr. Ayman Zawahiri’s recruits.

    Allow me the four graceful swans that eat the invasive seaweed that chokes the motors of passing boats.

  7. DXer said

    http://ieeexplore.ieee.org/xpl/login.jsp?tp=&arnumber=1167643&url=http%3A%2F%2Fieeexplore.ieee.org%2Fxpls%2Fabs_all.jsp%3Farnumber%3D1167643

    Directed killing of anthrax spores by microwave-induced cavitation

    Kiel, J.L. ; Air Force Res. Lab., Brooks, TX, USA ; Sutter, R.E. ; Mason, P.A. ; Parker, J.E.

    High-power pulsed-microwave radiation damages anthrax spores by apparent sonoluminescence in aqueous solutions containing the organic semiconductor diazoluminomelanin (DALM). DALM biosynthesized by JM109 E. coli, containing the plasmid pIC2ORNR1.1, had a higher affinity for spores of Sterne strain anthrax when compared to several other species of bacilli and enhanced the effect. Upon exposure to pulsed-microwave radiation, anthrax spores showed a maximum of 4 to 5 (i.e., 4.6) logs of kill. The light emitted was typical of plasma gas emissions and the spores, upon scanning electron-microscopic examination, showed enlargement and rupture typical of rapid expansion. Therefore, microwave-induced cavitations localized to the spore surfaces enhanced kill.

    Comment:

    Dr. Kiel did some experiments relating to the silicon signature of the anthrax mailed in Fall 2001. The lab found using silanizing solution in the slurry resulted in the same SEMS. But the silanizing solution did not enhance floatability — instead, it perhaps served some other purpose.

    This issue of “floatability” allowed the FBI to create a “straw man” — by knocking down a connection to floatabiilty they made it seem like they had knocked down the probative value of the signature.

    As a lay person judging expertise, I highly value the expertise of this Air Force lab. I think the scientists at that lab were experts had the pertinent insights — without the intentional obfuscation served up to the public on the issue (for understandable reasons).

  8. DXer said

    Belated congratulations to Steve Kurkjian on his movie deal. I’m a big fan of Mr. Kurkjian and the mystery — but am only free to join in the hunt during the summer after school’s out. There is a $5 million reward for the return of the paintings in good condition — to the extent only some are returned or the condition is not good, the reward would be pro rated.

    Off-topic:

    http://deadline.com/2015/04/master-thieves-matt-tolmach-tristar-1201418851/

    EXCLUSIVE: TriStar has pre-empted film rights for Pulitzer Prize-winning journalist Stephen Kurkjian’s Master Thieves: The Boston Gangsters Who Pulled Off The World’s Greatest Art Heist. Matt Tolmach will produce a film that will tell the story of the 1990 art heist at Boston’s Isabella Stewart Gardner Museum, where thieves took 13 works of art, including several Rembrandts and Vermeer’s The Concert. The stolen art was valued up to $500 million and some 25 years later, the case is still unsolved and the artwork still missing. Those works haven’t been forgotten: empty frames hang as a monument in the museum.

  9. DXer said

    Some history that is being overlooked in the reporting.

    Inadvertent Laboratory Exposure to Bacillus anthracis — California, 2004
    http://www.cdc.gov/mmwr/preview/mmwrhtml/mm5412a2.htm

    On June 9, 2004, the California Department of Health Services (CDHS) was notified of possible inadvertent exposure to Bacillus anthracis spores at Children’s Hospital Oakland Research Institute (CHORI), where workers were evaluating the immune response of mice to B. anthracis. This report summarizes the subsequent investigation by CDHS and CDC, including assessment of exposures, administration of postexposure chemoprophylaxis, and serologic testing of potentially exposed workers. The findings underscore the importance of using appropriate biosafety practices and performing adequate sterility testing when working with material believed to contain inactivated B. anthracis organisms.

    On May 28, 2004, CHORI staff members injected 10 mice with a suspension believed to contain nonviable vegetative cells of B. anthracis Ames strain. The suspension was centrifuged and drawn into syringes on an open bench in the laboratory. The mice were injected in a separate animal-handling facility at CHORI. By May 30, all of the injected animals had unexpectedly died. The carcasses were removed from the cages, placed into a plastic biohazard bag, and frozen. The bedding was discarded as standard animal waste. The cages were sanitized in an automated washer.

    On June 4, an additional 40 mice were injected with the same suspension. By June 7, all but one of these mice had died. All subsequent work was performed under a biological safety cabinet (BSC), and additional personal protective equipment (PPE) was used (e.g., protective clothing and gloves). Animal cages were brought into the BSC, and the surviving animal was euthanized. The carcasses were removed, placed into double biohazard bags, and frozen. The bedding and cages were autoclaved.

    On June 8, a sample of the original suspension was cultured; one mouse that died after the second experiment was necropsied and samples for cultures were obtained from its liver and peritoneal cavity. Within 24 hours, these cultures grew nonhemolytic gram-positive rods. Colony morphology was consistent with B. anthracis.

    Suspension material and cultures were transported to a California Laboratory Response Network (LRN) reference laboratory for further identification. The California LRN confirmed that the organisms isolated were B. anthracis by using polymerase chain reaction and gamma phage lysis assay. At CDC, antimicrobial susceptibility testing revealed that the isolates were susceptible to penicillin, ciprofloxacin, and doxycycline. Multiple-locus variable-number tandem repeat analysis confirmed that the isolates were genotype 62, consistent with B. anthracis Ames strain (1).

    On June 9, CDHS personnel visited the laboratory and animal-handling facility at CHORI to review the incident and laboratory procedures. No spills, puncture wounds, animal bites, or scratches were identified; however, initial handling of the suspension included snapping lids of microtubes, ejection of pipette tips, and centrifuging. The centrifuge tubes had snap-down tops, and the rotor was covered with a gasket. The laboratory procedures might have potentially expelled small drops of suspension but were considered unlikely to have released infectious aerosols. Because staff members believed they were working with inactive organisms, they had performed these activities on an open bench, and appropriate PPE was not consistently used until after the deaths of the second group of mice.

    As part of routine laboratory procedure, horizontal surfaces had been cleaned with a buffered bleach solution (1:10 dilution) at the end of each day. After laboratory workers recognized the possibility of exposure to viable B. anthracis spores, all laboratory surfaces and hoods were cleaned twice more with the bleach solution. The animal facility was also sanitized with bleach and a quaternary ammonium disinfectant.

    Twelve persons were involved in either the laboratory or its animal-handling facilities. Three of these persons had direct contact with the bacterial suspensions, cultures, or infected animals. Although at low risk for inhalation of B. anthracis spores, to further reduce their risk, the three workers with direct contact were recommended for postexposure chemoprophylaxis for prevention of inhalational anthrax (i.e., either ciprofloxacin 500 mg or doxycycline 100mg, orally twice daily for 60 days) (2). The nine persons who worked in the laboratory or animal-handling facility but who did not have direct contact were offered the same chemoprophylaxis regimen. All 12 were additionally offered, but declined, anthrax vaccine under an Investigational New Drug (IND) protocol for postexposure prophylaxis (3).

    Eight of the 12 potentially exposed persons opted to take chemoprophylaxis, including the three persons for whom the regimen was recommended. One person subsequently had a rash consistent with adverse reaction to ciprofloxacin; doxycycline was substituted. No other adverse effects from chemoprophylaxis were reported. None of the potentially exposed persons had symptoms consistent with anthrax.

    Serum specimens collected from nine (75%) of the 12 exposed persons 3–6 weeks after exposure were negative for IgG antibodies to B. anthracis protective antigen (PA) by enzyme-linked immunosorbent assay (4). Three persons did not provide sera for evaluation, including one person who had direct exposure to the bacterial suspensions and cultures.

    Further investigation revealed that the suspension had been prepared by a separate contract laboratory in March 2004 and contained an estimated 1.5 x 109 vegetative organisms per 1 mL of phosphate-buffered saline solution. After heating the suspension at 140ºF (60ºC) for 2 hours, the contractor reported that the suspension revealed no spores and had no growth after 48 hours of incubation on sheep blood agar.

    A sealed, screw-top tube containing the suspension was shipped to CHORI in a double-compartment package on wet ice and arrived intact. The tube of suspension was stored in a refrigerator until used. The suspension had been prepared specifically for the research laboratory and was not distributed to other facilities. All contractor laboratory personnel had received anthrax vaccine, and the suspension was prepared under biosafety level 3 (BSL-3) conditions.

    Leftover suspension from the incidents at the research laboratory were provided to CDC for quantification of viable organisms and to confirm the presence of B. anthracis spores. Sample dilutions were plated in duplicate on sheep blood agar. Approximately 2.0 x 106 colony-forming units (CFU) were enumerated per milliliter of suspension after 24 hours of incubation at 98.6ºF (37.0ºC). Comparisons of heat-shocked (149ºF [65ºC] for 30 minutes) and non–heat-shocked samples at CDC indicated that the suspension primarily contained B. anthracis spores.

    Reported by: A Lucas, PhD, Children’s Hospital Oakland Research Institute; M Doane, MD, J Rosenberg, MD, D Gilliss, MD, P Duffey, PhD, D Sesline, DVM, D Lindquist, MPH, R Das, MD, B Materna, PhD, D Vugia, MD, California Dept of Health Svcs. S Reagan, MPH, M Fischer, MD, N Marano, DVM, A Hoffmaster, PhD, V Semenova, PhD, S Martin, MT, C Quinn, PhD, Div Bacterial and Mycotic Diseases; J Patel, PhD, Div of Healthcare Quality Promotion, National Center for Infectious Diseases; M Kiefer, R Ehrenberg, National Institute for Occupational Safety and Health; R Weyant, PhD, Office of Health and Safety; B Ellis, PhD, T Jones, L Bane, M Hemphill, PhD, Office of Terrorism Preparedness and Emergency Response, Office of the Director, CDC.

    Editorial Note:

    The findings in this investigation indicate that workers in a research laboratory unknowingly received and used a suspension from a contract laboratory that likely contained viable B. anthracis organisms. Manipulation of the suspension at the research laboratory was determined unlikely to have expelled infectious aerosols, and exposed workers were considered at low risk for inhalation of spores. CDC continues to work with state agencies and other federal agencies to investigate processing procedures at the contractor facility to determine why the suspension contained viable B. anthracis organisms.

    B. anthracis spores are highly resistant to the effects of heat and chemical disinfection (5). Although the heat-killing procedures used by the contractor might have been lethal to vegetative cells, the procedures were not lethal to spores. Modifying suspension preparations by increasing the temperature and duration of heat-killing procedures or using formalin will increase the probability that spores are inactivated (5,6).

    Inactivated suspensions of B. anthracis should be cultured both at the preparing laboratory before shipment and at the research laboratory several days before use to ensure sterility. Sensitivity of sterility testing might be enhanced by increasing the inoculum size and incubation time, and by inoculating in multiple media, including both solid and broth media. Such procedures would increase the probability of detecting even a small number of viable B. anthracis spores. CHORI staff members did not perform sterility testing on the suspension received in March 2004.

    Because inhalation of viable B. anthracis spores can result in fatal infection, CDC recommends that laboratory personnel who routinely perform activities with clinical materials and diagnostic quantities of infectious cultures implement BSL-2 practices (7). These practices include use of appropriate PPE (e.g., gloves, gowns, or laboratory coats) and a BSC for procedures with the potential to expel infectious aerosols (e.g., centrifuging or ejection of pipette tips). Face protection (e.g., goggles, face shield, or splatter guard) should be used against anticipated splashes or sprays when potentially infectious materials require handling outside of the BSC. In the incidents described in this report, because CHORI staff members believed they were working with nonviable organisms, they did not fully implement BSL-2 practices until after the deaths in the second group of mice.

    Research laboratory workers should assume that all inactivated B. anthracis suspension materials are infectious until inactivation is adequately confirmed. BSL-2 procedures should be applied to all suspension manipulations performed before confirming sterility. After sterility is confirmed, laboratory personnel should continue to use BSL-2 procedures while performing activities with a high potential for expelling aerosolized spores.

    The Advisory Committee on Immunization Practices recommends routine anthrax vaccination of persons who work with production quantities or concentrations of B. anthracis cultures or perform other activities with a high potential for producing infectious aerosols (8). Facilities performing such work should have appropriate biosafety precautions in place to prevent exposure to B. anthracis spores; however, anthrax vaccination can be an additional layer of protection in the event of an unrecognized breach in practices or equipment failure. Because of the small potential for inadvertent exposure to aerosolized B. anthracis spores before or after sterility testing, vaccination might also be considered for researchers who routinely work with inactivated B. anthracis suspensions.

    In addition, laboratories working with inactivated B. anthracis organisms should develop and implement training activities and incident-response protocols to ensure appropriate actions are taken in the event of a potential exposure. These protocols should describe mechanisms for offering counseling and postexposure chemoprophylaxis and obtaining paired sera from potentially exposed persons. Training at animal research facilities should emphasize prompt communication between animal handlers and researchers if animals are unexpectedly found dead and any special handling procedures are needed for carcasses and bedding. Finally, institutional biosafety committees should routinely review protocols and procedures to ensure that appropriate safety precautions are always in place.

    References

    • Hoffmaster AR, Fitzgerald CC, Ribot E, Mayer LW, Popovic T. Molecular subtyping of Bacillus anthracis and the 2001 bioterrorism-associated anthrax outbreak, United States. Emerg Infect Dis 2002;8:1111–6.
    • CDC. Update: investigation of bioterrorism-related anthrax and interim guidelines for exposure management and antimicrobial therapy, October 2001. MMWR 2001;50:909–19.
    • CDC. Use of anthrax vaccine in response to terrorism: supplemental recommendations of the Advisory Committee on Immunization Practices. MMWR 2002;51:1024–6.
    • Quinn CP, Semenova VA, Elie CM, et al. Specific, sensitive, and quantitative enzyme-linked immunosorbent assay for human immunoglobulin G antibodies to anthrax toxin protective antigen. Emerg Infect Dis 2002;8:1103–10.
    • Turnbull PCB, Kramer JM. Bacillus. In: Murray PM, Baron EJ, eds. Manual of clinical microbiology. 8th ed. Washington, DC: ASM Press; 2003:349–56.
    • Spotts Whitney EA, Beatty ME, Taylor TH Jr, et al. Inactivation of Bacillus anthracis spores. Emerg Infect Dis 2003;9:623–7.
    • CDC, National Institutes of Health. Biosafety in microbiological and biomedical laboratories. 4th ed. Atlanta, GA: US Department of Health and Human Services, CDC, National Institutes of Health; 1999.
    • CDC. Use of anthrax vaccine in the United States: recommendations of the Advisory Committee on Immunization Practices. MMWR 2000;49(No. RR-15).

    Use of trade names and commercial sources is for identification only and does not imply endorsement by the U.S. Department of Health and Human Services.

    References to non-CDC sites on the Internet are provided as a service to MMWR readers and do not constitute or imply endorsement of these organizations or their programs by CDC or the U.S. Department of Health and Human Services. CDC is not responsible for the content of pages found at these sites. URL addresses listed in MMWR were current as of the date of publication.

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