* Dugway scientist Bruce Harper initiated shipment of the virulent Ames to USAMRIID for irradiation by email to Dugway colleagues dated June 21, 2001
Posted by DXer on October 22, 2015
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CASE CLOSED ... what really happened in the 2001 anthrax attacks? 
DXer said
Appl Environ Microbiol. 2001 Aug; 67(8): 3665–3670.
doi: 10.1128/AEM.67.8.3665-3670.2001
PMCID: PMC93069
Bacillus Spore Inactivation Methods Affect Detection Assays
Jessica L. Dang,1,* Karen Heroux,1 John Kearney,2 Ameneh Arasteh,3 Mark Gostomski,3 and Peter A. Emanuel3
Author information ► Article notes ► Copyright and License information ►
This article has been cited by other articles in PMC.
MATERIALS AND METHODS
Growth and processing of Bacillus cultures.
Geo-Centers, Inc., Lanham, Maryland 207061; Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 352942; and Research and Technology Directorate, U.S. Army Edgewood Chemical Biological Center, Aberdeen Proving Ground, Maryland 210103
*Corresponding author. Mailing address: Geo-Centers, Inc., Gunpowder Branch, P.O. Box 68, Aberdeen Proving Ground, MD 21010. Phone: (410) 436-8765. Fax: (410) 436-1912.
Inactivation and biodetection protocols were conducted using B. anthracis NNR1 (pX01+ pX02−), B. anthracis ΔAmes (pX01− pX02+), B. anthracis ΔSterne (plasmid free), and two negative controls, Bacillus subtilis strain 1031 and Bacillus thuringiensis subsp. kurstaki. Bacilluscultures were generously provided by Alvin Fox (University of South Carolina Medical School) and Bruce Harper (Dugway Proving Grounds, Utah).
DISCUSSION
The data demonstrate that inactivation methods can affect the sensitivity of nucleic acid-based detection and immunoassays for the detection of Bacillus spores. We have observed a differential effect based on the type of assay employed. The need to avoid handling and testing large amounts of potentially harmful spore preparations dictates that we understand the effects of inactivation procedures. We have studied two common spore inactivation procedures and demonstrated how they affect three types of biodetection assays. These effects should be taken into consideration when comparing laboratory results to data collected and assayed during field deployment.
***
The inactivation of weapons of mass destruction has implications for laboratory-based research and development of field-based applications of the various detection technologies. Future work should be aimed at platform- and method-specific testing of biological agents and the effects of decontamination on sensitivity and cross-reactivity. These issues are likely to become increasingly important in the realm of global politics, and further study is warranted.
Go to:
ACKNOWLEDGMENTS
We acknowledge the contribution of Peter Spaeth of SBCCOM for his technical assistance and expertise in sample irradiation, Patricia E. Anderson of Geo-Centers, Inc., for her technical consultation on flow cytometry, and Calvin Chue for his technical assistance with TaqMan PCR data.
This work was supported in part by DAAD 13-00-0-0015 (principal investigator, J. Kearney), MDA 972-96-K-003/P0003, and DAAD 19-00-1-0032.
DXer said
Why did an investigator bring Dr. Ivins a jar of GOLDEN EAGLE SYRUP upon a visit to Alabama?
Posted by Lew Weinstein on April 14, 2011
https://caseclosedbylewweinstein.wordpress.com/2011/04/14/why-did-an-investigator-bring-dr-ivins-a-jar-of-golden-eagle-syrup-upon-a-visit-to-alabama/
DXer said
John Ezzell was in charge of production of gamma-irradiation sterilized dried Ames and Sterne anthrax spores pre-911. (It was Ames and Sterne shipped on June 27, 2001). He worked for the FBI in collecting samples in Amerithrax and had worked for them since 1996 in a special unit that included some of the Amerithrax scientists. He or an assistant threw out Ivins’ submitted sample in February 2002.
Can Dr. Ezzell tell us what happened to the 340 ml. of virulent Ames? (And for that matter, the Sterne).
Both Ames and Sterne were involved in his dried powder project.
Was it in fact ever in fact irradiated? 340 ml. was an amount consistent with what would have been needed for the Fall 2001 anthrax mailings.
Our filmed Q and A a fews ago on this subject was interrupted when he had a heart attack was rushed to the nearby hospital during a break to have a stent put in.
I had gone to the restroom in a nearby building and my graphic artist had gone on an errand. By the time we came back, John had been taken to the hospital.
USAMRIID today produced an “Information Paper” explaining production of gamma-irradiation sterilized dried Ames anthrax spores pre-9/11
Posted by Lew Weinstein on July 26, 2014
https://caseclosedbylewweinstein.wordpress.com/2014/07/26/usamriid-today-produced-an-information-paper-explaining-production-of-gamma-irradiation-sterilized-dried-ames-anthrax-spores-pre-911/
DXer said
DXer said
Was this missing and unaccounted-for 340 ml. of virulent Ames the virulent Ames that was used for nefarious purposes to kill 5 people in Fall 2001?
Who can help out and make available the missing documents that are relevant?
“In May, 1999, the Fourth Edition of the BMBL was published. The Fourth Edition revises some of the biosafety standards contained in the Third Edition as a result of a number of events (e.g. emerging and re-emerging infectious diseases, laboratory associated infections and advances in facility design and construction). The Fourth Edition also contains biosecurity standards, which have not been included in previous editions of the BMBL. These biosecurity provisions are intended to assure that registered entities take measures to prevent unauthorized use of agents and/or use of agents by unqualified persons. These measures are also designed to protect against theft of these agents thereby decreasing the likelihood that the agents may be used for nefarious purposes.”
Requirements for Facilities Transferring or Receiving Select Agents
http://govpulse.us/entries/2001/08/31/01-22128/requirements-for-facilities-transferring-or-receiving-select-agents
On October 28, 1999, CDC published a Notice of Proposed Rulemaking (“NPRM”) seeking both to revise the biosafety standards facilities must follow when handling select agents and to provide new biosecurity standards for such facilities. These new standards are contained in the Fourth Edition of BMBL, which the NPRM proposed to incorporate by reference, thereby replacing the Third Edition. No comments were received in response to this proposal. CDC is therefore amending its regulations to incorporate the Fourth Edition.
For further information contact: ↑
Laboratory Registration/Select Agent Transfer (LR/SAT) Program, Office of Health and Safety, Centers for Disease Control and Prevention (CDC), 1600 Clifton Road, NE., MS-A13, Atlanta, GA 30333; telephone (404) 639-4418; LR/SAT Program website at http://www.cdc.gov/od/ohs/lrsat.htm.
Background ↑
“The Antiterrorism and Effective Death Penalty Act of 1996,” Pub. L. 104-132,(42 U.S.C. § 262 note) enacted on April 24, 1996, established new provisions to regulate the transfer of certain biological agents and toxins (i.e., select agents), and required HHS to issue rules to implement these provisions. The final rule was published in the Federal Register on October 24, 1996 (61 FR 551990-01) and became effective April 15, 1997. To comply with the final rule, commercial suppliers of select agents, as well as government agencies, universities, research institutions, and private companies that transfer these agents, must register with the Centers for Disease Control and Prevention (CDC). Prior to transferring or receiving a select agent listed in Appendix A of 42 CFR part 72, the facility must be equipped and capable of handling the covered agent at Biosafety Level (BSL) 2, 3, or 4, depending on the agent. The requirements for BSL 2, 3, or 4 operations are contained in the CDC/NIH publication “Biosafety in Microbiological and Biomedical Laboratories” (BMBL), and are currently incorporated by reference in the Third Edition. In May, 1999, the Fourth Edition of the BMBL was published. The Fourth Edition revises some of the biosafety standards contained in the Third Edition as a result of a number of events (e.g. emerging and re-emerging infectious diseases, laboratory associated infections and advances in facility design and construction). The Fourth Edition also contains biosecurity standards, which have not been included in previous editions of the BMBL. These biosecurity provisions are intended to assure that registered entities take measures to prevent unauthorized use of agents and/or use of agents by unqualified persons. These measures are also designed to protect against theft of these agents thereby decreasing the likelihood that the agents may be used for nefarious purposes.
Publication Date:
Friday, August 31, 2001
Document Number:
01-22128
Publishing Agency:
Department of Health and Human Services
Dates:
Effective date is January 1, 2002.
DXer said
Was this part of the ongoing work with biodetection?
Did Dr. Bruce Harper ship Ezzell the large quantities of Ames to USAMRIID in August 2000 and June 2001 to include a June 27, 2001 shipment that got misdirected?
Gamma-irradiated Ames was used at U.S. Army Edgewood under Bio-Safety 1 (BSL-1).
Did Dr. Bruce Harper send the Stern, Ames and Vollum to Dr. John Ezzell in August 2000 and June 2001 for Army-funded biodetection and decontamination experiments at Aberdeen in 2000 and 2001?
The flow cytometry of Bacillus anthracis spores revisited †
• Peter J. Stopa*
http://onlinelibrary.wiley.com/doi/10.1002/1097-0320(20001201)41:4%3C237::AID-CYTO1%3E3.0.CO;2-3/full
Article first published online: 13 NOV 2000
Work with gamma-irradiated spores of BA and other strains of Bacillus was performed at the U.S. Army Edgewood Chemical and Biological Center (ECBC) under Bio-Safety Level 1 (BSL-1) practices. Work that required the use of live BA spores was performed under Bio-Safety Level 3 (BSL-3) practices at the Baker Lifesciences Test Facility (BLTF), U.S. Army Dugway Proving Ground.
Gamma-Irradiated spores of BA (strains: Sterne, Ames, and Vollum) were obtained from Dr. John Ezzell of the U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID). …
Live spores of BA were provided by Dr. Lloyd Larsen (BLTF) for experiments that were conducted under BSL-3 conditions at Dugway Proving Ground.
…
For cross-reactivity studies, Bacillus globigii (BG; now known as Bacillus subtilis var. niger) was obtained from Dr. Bruce Harper (BLTF). BG was supplied as either a dry powder or as spore suspensions in water.
The U.S. Army Edgewood Chemical and Biological Center, 5183 Blackhawk Road, AMSSB-RRT-DD E3160, Aberdeen Proving Ground, MD 21010-5424 …
see also
TECHNICAL APPROACHES TO BIOLOGICAL AGENT DETECTION
PETER J. STOPA JOHN WALTHER US Army Edgewood Chemical Biological Center 5183 Blackhawk Road AMSRD-ECB-ENP-MC, E3549 Aberdeen Proving Ground, MD 21010-5424 USA
JEFF MORGAN
Applied Ordnance Technology, Inc. 25 Center Street Stafford, VA 22554
Peter Stopa noted that “Silica particles were (are) used to improve aerosol dissemination”
P. Stopa and Z. Orahovec (eds.), Technology for Combating WMD Terrorism, 53-66.
© 2004 Kluwer Academic Publishers.
DXer said
CDC should investigate inventory, shipping and radiation practices dating back to this June 27, 2001 shipment of this 340 ml. of virulent Ames which likely was genetically identical to the Ames that was used to kill five people in Fall 2001.
Anyone not part of the solution is part of the problem.
DXer said
In a secret report, Battelle determined that the sample provided by the FBI in October 2001 resembled the subtilis var. niger (“Bg”) surrogate made by Dugway
Posted by Lew Weinstein on October 13, 2015
https://caseclosedbylewweinstein.wordpress.com/2015/10/13/in-a-secret-report-battelle-determined-that-the-sample-provided-by-the-fbi-in-october-2001-resembled-the-subtilis-var-niger-bg-surrogate-made-by-dugway/