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

* Dugway anthrax irradiation scandal – USAMRIID’s Bruce Ivins in 2008 recognized that a 50% check of samples “leaves us ‘better safe than sorry’ ” (as opposed to other’s insistence that only 10% was necessary)

Posted by Lew Weinstein on June 13, 2015

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5 Responses to “* Dugway anthrax irradiation scandal – USAMRIID’s Bruce Ivins in 2008 recognized that a 50% check of samples “leaves us ‘better safe than sorry’ ” (as opposed to other’s insistence that only 10% was necessary)”

  1. DXer said

    Compare the Pentagon’s finding to Dr. Bruce Ivins’ observations and personal experience.

    Pentagon: Poor testing led to Army shipping live anthrax
    Alison Young and Tom Vanden Brook, USA TODAY
    http://www.usatoday.com/story/news/nation/2015/07/23/army-anthrax-shipments-pentagon-army/30154545/

    In the wake of the U.S. Army Dugway Proving Ground shipping live anthrax – by mistake – to dozens of labs, the Pentagon has released the report of its investigation.

    Even though the Army’s Dugway Proving Ground in Utah is the nation’s “largest producer” of inactivated anthrax for use in biodefense research, the Pentagon’s review found Dugway did far less testing than other U.S. military facilities to ensure potentially deadly specimens were killed before shipment to defense contractors and university and government labs. When Dugway used radiation to kill a batch of anthrax, it tested only 5% of the material to ensure it couldn’t still grow. This small sample size was “prone to error” and failed to detect that not all the anthrax spores had been killed, the report said.

    “The development and implementation of ineffective irradiation and viability testing procedures took place over the last decade,” the report said, adding that “this represents an institutional problem at [Dugway] and does not necessarily reflect on any one individual.”

    The report is the result of a comprehensive review ordered May 29 by Deputy Secretary of Defense Bob Work to analyze the failures of processes to kill anthrax specimens before they were shipped.

    “By any measure, this was a massive institutional failure,” Work said at a news conference Thursday afternoon.

    The military’s ongoing investigation has determined that Dugway sent live anthrax specimens to at least 86 labs operated by companies, academic institutions and government agencies for use in developing tests and equipment to protect against bioterror attacks. Those labs are in the USA and seven foreign countries: Japan, South Korea, the United Kingdom, Australia, Canada, Italy and Germany. Though no illnesses have been linked to the specimens, at least 21 people who had direct contact with the specimens have been given antibiotics as a precaution.

    ***

    The Pentagon review found that no other military facilities involved in the production and inactivation of anthrax research specimens appear to have mistakenly shipped any live anthrax. Unlike at Dugway, the report said, staff at other military labs were able to routinely identify when they had live anthrax in their irradiated samples.

    ***

    Dugway scientists used Cobalt 60 gamma radiation to kill or deactivate anthrax specimens before shipping them to government or private labs for further research, the CDC report said. Dugway’s standard procedures for irradiating anthrax “did not account for the variable amounts of spores treated in the gamma cell irradiator,” the report said. The method used “was not validated using standardized control spore samples at varying concentrations, volumes, and levels of irradiation.”

    The Army’s own anthrax scientists have apparently been aware for years that the military’s methods for killing anthrax specimens — then doing tests to verify the kill step was fully effective — were not trustworthy. More than seven years ago, the nation’s most infamous anthrax researcher warned military colleagues about the issue. The emails of Bruce Ivins, the Army microbiologist accused in the anthrax letter attacks in 2001, raised concerns that when a batch of anthrax was irradiated that too few samples were tested to ensure all the spores were dead, USA TODAY reported last month. In a 2008 email, Ivins discussed how some think only 10% of an irradiated batch needed to undergo sterility testing, but he’d found at least 50% should be tested because of past irradiation failures.

    ***

    At Edgewood, staffers test 10% of an irradiated anthrax batch to check for any live bacteria, the report said, noting that this is “insufficient for verification of complete inactivation.” At USAMRIID, the review found staff tested 10% of an irradiated anthrax batch but noted that a researcher at the facility doing other spore work “chose to conduct viability testing by taking 50% of the preparation as the sample.”

  2. DXer said

    Army lab lacked effective anthrax-killing procedures for 10 years
    Alison Young, USA Today, June 17, 2015

    The U.S. Army research facility that has mistakenly shipped live anthrax to unsuspecting labs in the U.S. and abroad for more than 10 years failed to have effective and standardized procedures for killing the deadly bacteria with radiation, according to a federal investigation report obtained by USA TODAY.

    The report, dated June 5, cites the Dugway Proving Ground’s Life Science Test Facility in Utah with three violations of federal regulations for working with potential bioterror agents and orders the facility to immediately cease all shipments of “inactivated” anthrax specimens.

    According to the report by lab inspectors at the Centers for Disease Control and Prevention, Dugway scientists were using Cobalt 60 gamma radiation to kill or deactivate anthrax specimens before shipping them to government or private labs for further research.

    But Dugway’s standard procedures for irradiating anthrax “did not account for the variable amounts of spores treated in the gamma cell irradiator,” the report noted. The method used “was not validated using standardized control spore samples at varying concentrations, volumes, and levels of irradiation.”

    As a result, anthrax bacteria were shipped out at least 74 times to dozens of labs in the U.S. and at least five foreign countries from January 2005 to May 2015. Anthrax spores can be potentially fatal if inhaled.

    ***

    The new CDC report about Dugway’s anthrax mishaps, which is only three pages long, provides little detail about radiation dosages and durations, and exactly how Dugway’s scientists were verifying that each batch was fully killed. The report addresses the problems in general terms.

    ***

    The emails from accused anthrax letter terrorist Bruce Ivins — a microbiologist at the Army’s elite infectious disease laboratory in Fort Detrick, Md. — offer possible clues about what may have happened at Dugway.

    Emails sent by Ivins during the normal course of his work with anthrax at the U.S. Army Medical Research Institute of Infectious Diseases, often called USAMRIID, provide a window into the difficulties scientists face killing the bacterium and its hardy spores. They also indicate that scientists did not follow a universal, standardized protocol for what percentage of anthrax specimens in an irradiated batch needed to undergo verification tests before the batch was considered sterile and safe for shipment and use without significant safety precautions.

    The emails are among a massive trove of Ivins’ correspondence released in 2010 and posted online in a Freedom of Information Act reading room of the U.S. Army Medical Research and Materiel Command after federal officials formally closed their investigation into the 2001 anthrax attacks. Ivins, 62, died from an intentional overdose of acetaminophen in July 2008 as prosecutors prepared to charge him with sending the anthrax-filled letters that killed five and sickened 17 others.

    In a March 2008 email about the planned shipment of some irradiated anthrax spores, Ivins references a recent division meeting and wrote: “As discussed at the meeting, 50% (5 ml) of the material was checked for sterility on SBA, and there was no growth at all. You may have seen [redacted] insistence that we need only check 10% of the material for sterility, but I think that a 50% check for B. anthracis samples is a good idea. We’ve had in the past some samples that failed sterility checks, so 50% leaves us ‘better safe than sorry.’ ”

    The email indicates that to verify that the irradiated spores were killed, samples were being put on a sheep blood agar plate to see if the bacteria would grow.

    It is unclear whom Ivins was writing to in the email or where the specimens were going to be sent. The Army has redacted all other names, including that of the person or organization Ivins said was advocating only testing 10% of the irradiated samples. Over the years, Ivins’ emails show he did work with anthrax researchers at Dugway Proving Ground and that USAMRIID had an anthrax spore production contract with Dugway.

    In 2006 and early 2007, Ivins’ emails indicate that he did verification tests only on 10% of irradiated anthrax specimens. “The spores were irradiated on October 30. On November 2, [redacted] plated out 10% of each of the preps onto SBA plates. All were negative for growth 24 hours later. They will be incubated over the weekend,” Ivins wrote in a Nov. 3, 2006, email.

    By the summer of 2007, however, Ivins and his colleagues ran into a concerning series of irradiation failures, the emails show.

    “The 27 spore samples that were irradiated on 30 MAY 07 (see enclosed file) came back still ‘hot,’ ” Ivins wrote in a June 7, 2007, email sent to eight other people whose names are redacted. The specimens, he wrote, would be sent back for another irradiation dose of 1 million rads “which should kill the remaining viable spores.” Emails in the string show the May irradiation dose was 5 megarads.

    But Ivins had bad news to report on June 13, 2007: “Irradiation sterilization failure … again,” said the subject line of his email, which said that even with the additional radiation dose, the spores could still grow. His email said that verification tests were done on 18 of the 27 samples — and 12 of the 18 were still “hot.” And so they were again going to be sent back to receive an additional 2 megarads of radiation. “Hopefully this will work. Stay tuned … – bruce,” wrote Ivins

    An unidentified recipient of the email replied: “What is going on with the irradiator? Is it not working properly? These things are going to be fried to a crisp!”

    Ivins was puzzled, too. “In the past, 4 megarads would do the job. This is 2 million rads over that, and still no sterility,” he replied.

    The next morning, June 14, 2007, Ivins emailed with more bad news: “After recheck of the 6 samples that appeared to pass sterility check after a total of 6 million rads, only two samples remained negative.” The other four, it turned out, weren’t dead and needed more radiation, he wrote.

    It’s unclear what may have caused the repeated irradiation failures. Issues relating to irradiation are again mentioned in the publicly available emails in August 2007, when an unidentified person wrote to Ivins asking for the paperwork on the 27 spore samples. In his reply, Ivins wrote that “several rounds of irradiation had to be done to finally get all of the spores sterile. It seems as though the new irradiator isn’t as reliable as the old one for some reason.”

    Military officials didn’t answer USA TODAY’s questions about the radiation doses used at the Dugway Proving Ground lab, which is the focus of the current international investigation of live anthrax shipments. They also didn’t answer questions about what percentage of anthrax specimens in each irradiated batch were undergoing verification tests for sterility at Dugway. They did, however, email a statement on Wednesday about Ivins’ emails detailing the 2007 irradiation failures:

    “It is clear from these emails that the investigators did their due diligence to determine what was causing the failure and that no live material was used in the lab or sent to other labs,” said the statement emailed by Maj. Eric Badger, a Defense Department spokesman. As part of the department’s comprehensive review of Dugway’s mishaps, investigators are “examining, among other things, the failure rates of gamma irradiation for killing anthrax.”

  3. DXer said

    Appl Environ Microbiol. 2008 Jul; 74(14): 4427–4433.
    Published online 2008 May 30. doi: 10.1128/AEM.00557-08
    PMCID: PMC2493171
    Gamma Irradiation Can Be Used To Inactivate Bacillus anthracis Spores without Compromising the Sensitivity of Diagnostic Assays▿

    Leslie A. Dauphin, Bruce R. Newton, Max V. Rasmussen,† Richard F.
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2493171/

    The use of Bacillus anthracis as a biological weapon in 2001 heightened awareness of the need for validated methods for the inactivation of B. anthracis spores. This study determined the gamma irradiation dose for inactivating virulent B. anthracis spores in suspension and its effects on real-time PCR and antigen detection assays. Strains representing eight genetic groups of B. anthracis were exposed to gamma radiation, and it was found that subjecting spores at a concentration of 107 CFU/ml to a dose of 2.5 × 106 rads resulted in a 6-log-unit reduction of spore viability. TaqMan real-time PCR analysis of untreated versus irradiated Ames strain (K1694) spores showed that treatment significantly enhanced the detection of B. anthracis chromosomal DNA targets but had no significant effect on the ability to detect targets on the pXO1 and pXO2 plasmids ofB. anthracis. When analyzed by an enzyme-linked immunosorbent assay (ELISA), irradiation affected the detection of B. anthracis spores in a direct ELISA but had no effect on the limit of detection in a sandwich ELISA. The results of this study showed that gamma irradiation-inactivated spores can be tested by real-time PCR or sandwich ELISA without decreasing the sensitivity of either type of assay. Furthermore, the results suggest that clinical and public health laboratories which test specimens for B. anthracis could potentially incorporate gamma irradiation into sample processing protocols without compromising the sensitivity of theB. anthracis assays.

    ****

    A review of B. anthracis spore inactivation methods by Spotts Whitney et al. (38) listed gamma irradiation as a method for inactivating B. anthracis spores. Horne et al. (16) described the use of gamma radiation for inactivation of virulent B. anthracis spores and found that a dose of 1.5 × 106 rads was required to inactivate live spores at a concentration of 106 spores/ml. A more recent study conducted by Dang et al. (7) reported that 2.0 × 106 to 2.24 × 106 rads was sufficient to inactivate B. anthracis spores at a concentration of 108 CFU/ml, but the study used only avirulent strains missing one of the two virulence plasmids. In addition, the study (7) reported that irradiation decreased the sensitivity of real-time PCR and antigen detection by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) and polyclonal antibodies (PAbs).

    The purposes of this study were to determine the gamma radiation dose for inactivating virulent B. anthracisspores in suspension and to characterize the effects of gamma irradiation on the limit of detection for real-time PCR and antigen detection assays. Experiments were carried out using virulent strains representing eight genetic groups of B. anthracis, while testing employed a validated real-time PCR assay currently used by LRN laboratories (15) and three ELISAs.

    ***

    This study is the first to report the gamma radiation dose for inactivating live virulent spores representing eight described genetic groups of B. anthracis (22)

    _______________

    *Corresponding author. Mailing address: BRRAT Laboratory, DBPR, NCPDCID, CDC, Mail Stop G-42, 1600 Clifton Road, Atlanta, GA 30333. Phone: (404) 639-4922. Fax: (404) 639-4234. E-mail: VOG.CDC@6BKM
    †Present address: Biosciences Defense Division, Lawrence Livermore National Laboratory, 7000 East Ave., L-452, Livermore, CA 94551.
    ‡Present address: The Tauri Group, 675 N. Washington St., Suite 220, Alexandria, VA 22314.

    • DXer said

      Whitney EAS, Beatty ME, Taylor TH Jr, Weyant R, Sobel J, Arduino MJ, et al. Inactivation of Bacillus anthracis spores. Emerg Infect Dis [serial online] 2003 Jun [date cited]. Available from: URL:http://www.cdc.gov/ncidod/EID/vol9no6/02-0377.htm

      Emerg Infect Dis. 2003 Jun;9(6):623-7.
      Inactivation of Bacillus anthracis spores.
      Spotts Whitney EA1, Beatty ME, Taylor TH Jr, Weyant R, Sobel J, Arduino MJ, Ashford DA.
      Author information

      Abstract
      After the intentional release of Bacillus anthracis through the U.S. Postal Service in the fall of 2001, many environments were contaminated with B. anthracis spores, and frequent inquiries were made regarding the science of destroying these spores. We conducted a survey of the literature that had potential application to the inactivation of B. anthracis spores. This article provides a tabular summary of the results.
      PMID: 12780999 [PubMed – indexed for MEDLINE] PMCID: PMC3000133

      http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3000133/

      Gamma radiation was used in the 1960s and 1970s to disinfect B. anthracis–contaminated imported bailed goat hair. A study by Horne et al. suggested that a dose of 1.5 megarads from a 200,000-rad/hour cobalt source was sufficient to kill most resistant spores when mixed with goat hair; 2 megarads was recommended to include a margin of safety (31). After the intentional release of B. anthracis through the postal system in 2001, pursuing a decontamination method for the undelivered mail was essential. Gamma radiation was used to decontaminate all mail from contaminated facilities on the basis of these data.

  4. DXer said

    During this period, JAG was refusing to let Bruce Ivins’ attorney speak to his co-workers. Defense counsel never had an opportunity to point out the unvalidated premise of the FBI’s genetic analysis — the FBI’s assumption that all irradiation or inactivation by other means was successful.

    From:
    To:
    Subject:
    Date:
    Ivins, Bruce E Dr USAMRIID
    JAG permission for my attorney to talk to USAMRIID personnel (UNCLASSIFIED) Thursday, May 29, 2008 11:48:25 AM
    (b)(6)

    Classification: UNCLASSIFIED Caveats: NONE

    Dear

    I am sending you this email to ask if you had heard from the JAG as to when my attorneys, , of , can talk to USAMRIID personnel about me. If you have not yet heard, could you please contact them and ask them? Then I can let my attorneys know the information. I’ve been a loyal Army employee for over 27 years, and I would hope that the Army would similarly have a substantial measure of loyalty toward its employees. Thank you very much.

    -Bruce
    Bruce Ivins

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