Monday, August 25, 2008

Science Magazine/ ANTHRAX INVESTIGATION: FBI Discusses Microbial Forensics--but Key Questions Remain Unanswered

Science 22 August 2008:
Vol. 321. no. 5892, pp. 1026 - 1027

excerpted from the body of the article: (my apologies that a free link is not available)

...The FBI disclosed earlier that only eight samples had all four mutations; on Monday, it said that all but one of these came from USAMRIID. And all eight could be traced to RMR-1029--the flask of spores under Ivins's charge.

That is as far as the science took them, the FBI conceded; conventional detective work--such as checking lab notebooks and shipment records--helped rule out everyone but Ivins who had access to the spores, says Vahid Majidi, head of the agency's Weapons of Mass Destruction Directorate. He declined to give details. "I'm asking you not to second-guess our investigative approach," he said...

But Science was no more able to avoid second-guessing the FBI investigation than the rest of us. The journal came up with additional questions for the FBI:

Six Anthrax Science Questions the FBI Has Yet to Answer

Yudhijit Bhattacharjee and Martin Enserink

  • What were the four mutations the FBI says it used to link the anthrax in the envelopes to Bruce Ivins at USAMRIID?
  • What are the odds of a false positive--that is, the odds that the spore populations in Ivins's flask RMR-1029 and in the envelopes weren't related but shared the same four mutations by chance?
  • Eight samples had anthrax with all four mutations; one of those came from a lab other than USAMRIID. On what basis was this lab ruled out as the origin of the letters?

  • How did the FBI rule out the possibility that others at USAMRIID with access to Ivins's lab prepared the envelopes?
  • How exactly did Ivins, if he was the perpetrator, produce an easily dispersible powder from his anthrax culture?
  • What led the FBI to suspect Steven Hatfill in the earlier years of the investigation?

    Anonymous said...

    Well, being an older fellow and inclined to discount all this fancy mutational rigmarole, I wonder what happened to old methods of investigation.

    Have all the handwriting analysts gone bye-bye, or is that now a discredited technique?

    If, as the FBI conceded, they've been "checking lab notebooks and shipment records," why then no comparison pro or con of Ivins's known handwriting and that on the envelopes?

    Or are we to infer that decades worth of cases where handwriting analysis helped nail the perpetrator are now of questionable resolution?

    The poor guy had been filling out "shipment labels" for decades, and now (2001) here appear hand-addressed lethal envelopes. Surely the FBI looked, compared Ivins's shipment records and the anthrax envelopes, right? Did diabolically clever Ivins's teach himself to fool the experts? Did an assistant write (print) for him?

    Please, for us of the old Sherlock Holmes school, somebody tie a bow around the handwriting.

    Anonymous said...

    These are excellent questions, especially how the unnamed lab came to be ruled out, and the odds of false positives.

    Another thing I have always wondered is if the pressure and possible electrostatic charge of the postal machines themselves helped mill/charge the spores in the envelopes. (In addition to sending them floating around the processing centers.) Seems like the FBI would have tested this, but you never know.

    One other hole in the investigation is the death of Cathy Nguyen, the Bronx hospital worker who died of inhalation anthrax. As far as I know, the investigation has not been able to place her near any of the mailings.

    Anonymous said...

    I'm asking you not to second guess our invetigative approach.

    Who is scripting the government, Monty Python?

    It's an absurd thing to say, especially after Hatfill.

    and those documents linking Ivins, supposedly, to a school in Wisconsin, and the "kiss my ass" journalist, and, and, These are so embarassing the biggest critics won't talk about it. I hope Congress does.

    Anonymous said...

    "That is as far as the science took them, the FBI conceded; conventional detective work--such as checking lab notebooks and shipment records--helped rule out everyone but Ivins who had access to the spores . . ."

    So, they don't know who had access that they don't know about. Is that right?

    And, am I mistaken or is Majidi asking people not to think about their methods?

    "I'm asking you not to second-guess our investigative approach," he said...

    daedalus2u said...

    How were these mutations sampled? My understanding is that the spores were cultured and then colonies were picked by hand. There is a large degree of subjectivity in which colonies to pick. Unless enough were taken to get a statistical sample, and it doesn't seem as though enough were.

    Anonymous said...

    This is a little off topic for this thread but look at this:

    August, 6, 2008: Jeff Taylor: But again, back to circumstantial evidence — thousands of prosecutors in thousands of courthouses prove cases beyond a reasonable doubt using circumstantial evidence. In fact, the standard jury instruction given by judges across the country is that a jury can consider circumstantial evidence and direct evidence, and they both can be given equal weight depending on the jury's view. So, again, circumstantial evidence? Sure, some of it is. But it's compelling evidence and our view is we are confident it would have helped us prove this case against Dr. Ivins beyond a reasonable doubt.

    Daschle said the most compelling evidence to him is the odd, extended hours that the Army scientist kept shortly after the Sept. 11 terror attacks.

    New York Times: In its case against Dr. Ivins, the F.B.I. developed a compelling profile of an erratic, mentally troubled man who could be threatening and obsessive, as in his odd fascination with a sorority from his college days. But investigators were never able to place him at the New Jersey mailboxes where the anthrax letters were dropped, and the case against him relied at its heart on the scientific evidence linking the anthrax in Dr. Ivins’s laboratory to the spores used in the attacks.

    "I am persuaded, unless I'm missing something, there is a compelling case they at least got the one right guy," Smith said. "They claim there's no evidence whatsoever that there was an accomplice, but our hope is that they still keep looking to make sure there wasn't."

    8/8/AP: Mark Cunningham, a New York Post op-ed editor, one of three staffers there who were sickened by an anthrax-tainted letter, said he also was convinced about the government's case against Ivins.

    "The case is circumstantial but compelling," Cunningham wrote in a column published on the paper's Web site yesterday. "I'm glad they're keeping the case open, to tie up loose ends, make absolutely certain he acted alone, and all the rest. But I have my closure.",0,6321499.story?track=rss

    "The scientific evidence is compelling," says Rita R. Colwell, former director of the National Science Foundation, which funded some of the research behind the investigation. It is impressive how all the different scientific aspects came together, she says.

    Nancy Haigwood:

    Haigwood said FBI agents were "very ethical and above board." And reading their case files convinced her they have the right suspect. "The evidence was compelling," she said.

    Anonymous said...

    "I'm asking you not to second-guess our investigative approach," [Vahid Majidi, head of the FBI's Weapons of Mass Destruction Directorate] said.

    Mr. Majidi needs to take another look at the US Constitution to which he swore allegiance. In the United States, every citizen is presumed innocent until proven guilty. Indeed, it is both the right and duty of US Citizens to demand that Mr. Majidi back up his contentions with facts.

    Based on the dearth of evidence produced by the FBI to date, one has to wonder whether Mr. Majidi's choice of the term "second guess" was a Freudian slip. The FBI has no hard evidence and very little circumstantial evidence establishing Dr. Bruce Ivins as the Anthrax killer. As such, it is apparently the FBI who is in fact "second guessing" in this case, (Dr. Hatfill was, after all, the FBI's "first guess").

    Anonymous said...

    Someone posted earlier about Cathy Nugyen.
    I can 100% guarantee that she received a letter addressed to her and that Otillie Lundgren did as well. I know because I also received a letter and I have something in common with both of these women. We were in the same internet chat room at the same time. When I told Special Agent Jim Page at the Raleigh FBI about this he said "that's just a coincidence".
    So for me, having this inside information and the FBI falling over themselves not to investigate it, even though I gave Agent Eric Davis at the Charlotte FBI, Hatfill's last name in November of 2001, 7 months before anyone ever heard of him, for me it is plain as day that the FBI never had any intention of actually solving this case and they still have not solved this case.
    What they have done is pay one of the major culprits a little more than 1 million dollars for each person he killed. Which then in turn leads me to believe that members of the DOJ and FBI were in on it from day one.

    Anonymous said...

    Anonymous, what kind of chat room was it and what made you mention Hatfill's name? Did your letter contain anthrax?

    Old Atlantic Lighthouse said...

    Elizabeth Ferrari's list is compelling that this case needs better evidence and more disclosure and that the only thing closed is the FBI's mind.

    The FBI also seems to have become overly fixed on the Princeton NJ mailbox. They claim it was mailed there both times. But really there is no proof it was mailed there either time.

    If the letters had 2g of anthrax each, and each gram had a trillion spores that is 2 trillion spores inside. If there was one millionth of a gram on the outside of the envelope, that is 1 million spores on the outside. 8 to 10 thousand spores is a lethal dose. 125 spores can be detected by a swab. The postal carrier who picked up the letters should have gotten sick. The following table indicates some people who got sick. The FBI should figure out if one of the pick up people got sick and what boxes they picked up letters from.

    The following has a good table of the onset of anthrax in some of the postal workers and other victims.

    The FBI wants the Nassau St mailbox to be the one, so they can fix it to the sorority. But for the first mailing, if he drove up on Monday Sep 17 when he had leave he could have dropped it in some other box that picked up mail at 9 AM only so that it would be picked up on Tuesday. The Nassau St. mailbox could have been cross contaminated in this scenario. He could have picked a mailbox that was isolated so he could use plastic gloves.

    The FBI doesn't appear to really think straight. This is a reflection of not sharing information even internally. They need to put up charts on the wall and internet that map out every step of what they think happened where and when. Then link those to evidence. They also need to do little probability calculations, even crude ones for each event and all of them together.

    Anonymous said...

    It sounds a lot like they were entirely subjective in their choice of mutations. They intentionally chose colonies that looked different in order to find differences when they did the sequencing. That's not a really a big deal. The thing we'd like to know is how common the individual mutations are in FBI's 1000-sample archive. That would give us an idea how statistically useful the mutations are. It may be that they're common enough that a couple in a thousand samples of anthrax might have all four by chance. That doesn't seem likely, though.

    A couple of things have been bothering me about this:

    * Microbiologists, at least nowadays, try like mad to avoid working in mixed culture. We prefer working in pure lines, doing several passages in order to get to that point. This is especially true considering that we want to find interesting biochemistry and sequences, and it's harder to get clean data if the cultures themselves are dirty. Ivins did the exact opposite: he blended spore preps into a huge batch that he knew was a mixed population. Even if he hadn't known at first, he would have found out as soon as he subcultured the batch and found at least four colony types. That may have something to do with the immunology he did, but I can't speak to that. I don't see why anybody else would have wanted a subsample of this stock except as a curiosity.

    *When FBI requested those 1000-odd samples from those 16 labs, it seems likely that very few would be from a mixed stock for the above reason.

    *The odds that a pure anthrax culture would have all four mutations is probably very small, but the odds of that a mixed population would have all four is higher, provided you can get samples from people working in mixed cultures. It seems to me that they probably could trace the other USAMRIID samples back to Ivins, if only because it's not likely that anybody else would have a mixed stock. It doesn't say a thing about how they cleared everybody else who had access to any of the seven stocks held there.

    *We know nothing at all about the sample submission protocol. Kemp claims that the second submission was a misunderstanding, and that Ivins thought FBI wanted a pure culture. I wonder how many other scientists would have unthinkingly made that same mistake. Subculturing bacteria is almost invariable done from single, isolated colonies, not from an entire plate, and that's especially true for subcultures meant for genetic analysis. Written protocol to the contrary, the submittors may simply have followed their normal isolation procedures out of thoughtlessness or sloppiness. If they found several colonies, they may have sent sample A1, A2, A3, etc. despite the protocol.

    Ennealogic said...

    Then, there's the issue of the anthrax letter that killed Bob Stevens, the first victim. According to a June, 2002 Newsmax article:

    "About a week before Sept. 11, 2001, a letter arrived at AMI. It ended up in the offices of the Sun, one of AMI's tabloids. People at the Sun who saw it said it was a "weird love letter to Jennifer Lopez." Inside the letter was a "soapy, powdery substance" and a cheap Star of David charm."

    Note -- a week before 9/11/2001.

    "The letter, which had been taken to the Sun by Ernie Blanco, was passed around the office for its amusement value and ended up on Bob Stevens' desk, where he appears to have inhaled the powdery substance. Stevens then threw the letter away in the trash."

    The article goes on to say:
    "Because the letter that appears to have been the source of the anthrax at AMI no longer existed, a vital piece of evidence was lost. That the letter was the source is indicated by the fact that the trail of anthrax spores in the AMI building matches the exact route it took from the mailroom to the Sun tabloid office."

    Since the FBI infers, in the affadavits they released in support of search warrants for Ivins' property, that spores from all FIVE deceased victims match the anthrax in flask RMR-1029, what effort did they make to identify where the Stevens letter was mailed from and when? Why was the content of the letter different (soapy powder, goofy love letter)? How does Ivins tie in with this first victim? Surely if he was the sole perpetrator, there would be a motive (and means and opportunity) to produce and mail this anthrax too. Was this the first of three batches of anthrax, since it clearly pre-dates the others by more than a week?

    Curiously, the government provided absolutely no assistance to AMI in the cleanup of their building. Curiouser still, after the building was abandoned and quarantined for a couple of years, a businessman bought it for $40,000 -- it had been worth $34 million. Then, Rudy Giuliani started a decontamination company and got the contract to clean the deserted AMI building of residual anthrax. Once decontamination was complete, the owner sold the building for over $90 million.

    Small world. More 9/11 connections. Star of David charms. This case keeps getting stranger and stranger.

    Anonymous said...

    They are refuting that the anthrax killer ever mailed a weaponized form of the strain:

    Anonymous said...

    "Anonymous, what kind of chat room was it and what made you mention Hatfill's name? Did your letter contain anthrax?"

    It was a major network news message board.
    I gave agent Eric Davis Hatfill's last name because he was walking down my street gawking at my house. He was wearing his army jacket which had his last name printed on it.

    Anonymous said...

    "Anonymous, what kind of chat room was it and what made you mention Hatfill's name? Did your letter contain anthrax?"

    Forgot to answer the last question. The letter I received said "We feel the US should not bomb Afghanistan for the WTC attack" IT was loaded with white powder that blew up on me out in the yard where I opened it. That was Tuesday after Columbus day. Later that evening a spot on my arm itched like nothing I have ever felt before. Over the next few days the spot grew larger and eventually started to get black in the center. It resembled a brown recluse bite. On the following Saturday the news was talking about Bob stevens and that it may have gotten him through the mail. I remembered the letter and went to the Urgent Care. They said they could not help and to go to the hospital. So I did. What a fiasco The hospital called the SBI who referred them to the FBI which is the first time I spoke with S.A. Jim Page. Long story short they took throat and nose swabs and said they were not going to swab the actual infection site because it would be sent to the lab. Guess what. When the doctor cuts a sample that big off your arm he sterilizes it first. So the State lab not the CDC (none of my samples have ever been sent to the CDC as S.A. Jim Page informed me) said it was negative for Anthrax. Later our Gov. Mike Easley made an announcement that all future test for Anthrax would not be done by the state because they had issues getting accurate results. This came after it was leaked that the Westgate postal facility which handles all NC mail tested positive for Anthrax. I do not have the letter but I do still have powder from it that will test positive if anyone would ever test it. I have offered to pay for testing. NO ONE will test it. I have a later from the FBI stating "Since his medical diagnosis failed to substantiate his allegations, no further investigation was conducted. We are, therefore, unable to be of further assistance to him."
    Arthur Radford Baker
    Unit Chief
    Office of Public and Congressional Affairs.

    As I said I still have powder from that letter which is evidence.

    Meryl Nass, M.D. said...

    That is a very interesting story. If it is true, I will do my very best to get your powder investigated.

    I assume it is encased in multiple plastic bags? And the outer bag will have been washed on the outside?

    Send me a copy of an ER record or other record confirming the story if you would like my help with this. The analysis will need to be done at a lab(s) authorized by the states to perform this type of analysis, or be done at the federal level.


    Anonymous said...

    "Send me a copy of an ER record or other record confirming the story if you would like my help with this."

    Thanks, I will send you an email. I must scan them in first.

    Anonymous said...

    Anonymous, I did a little Googling and I found the story you relate as far back as May of 2007 (although told in more of a third person style in the earliest post I found.)

    I have a couple of questions for you--other than the obvious "Dude, is this true?"

    Have you ever written your story from beginning to end? I mean starting with "My name is....and I live is what happened...and that's the whole story." If not, I hope you will do so, if only to avoid the confusion of telling it in bits and pieces.

    What was the major network news chat room and how on earth did you end up posting there with Ottile Lundgren and Cathy Nguyen? Lundgren was over 90 and one would not expect her to be a big internet user.

    Did you know Hatfill prior to the time you saw him near your house, and do you have any connection with the bioscience community? For that matter, do you know that the person you saw was Hatfill? (i.e. did you subsequently identify him in photos as the "Hatfill" you saw near your home?)

    For Dr. Nass, would there be any indication of previous anthrax exposure if anonymous were to be tested now? (I am trying to think of ways to corroborate this story which, while odd, seems no less farfetched than some details of the Ivins narrative.)


    Anonymous said...

    Like I try to correct others , correct me please if I’m wrong:

    Anonymous 27-8-08/9:03 PM: The misunderstanding about weaponized anthrax started when scientists were perplexed by the quality and pureness of the (second wave) samples and called it “weaponized”- quality. Silicon was found, not silica (let alone bentonite) as was claimed by some.
    Ennealogic: The J-Lo letter was positively NOT the one that infected Bob Stevens:

    Unknown said...

    Dr. Nass, please post updated information relative to Anonymous' claims on your blog. This seems incredible to me if true. Clearly if you get credible evidence from Anonymous (hospital info, letters and clearly powder) we'd certainly want to know about it. It all sounds too bizarre to me, but that would be consistent with the rest of this entire fiasco. thanks.

    Anonymous said...

    re: August 28, 2008 4:32 AM

    Okay, so the part where people got to thinking it was weaponized came from the period when the MSM were linking it to Iraq weapon's programs and it was sold as weaponized.

    But it was not weaponized, is that right? I had been under the impression it was weaponized and that this ruled Ivins out because it would have had to have been processed elsewhere by a sophisticated process.

    So, technically could it have been done by Ivins? Was he capable then or was the milling outside of his know-how or his access to the right equipment? Dr. Nass could you comment on this?

    Anonymous said...

    Here's my layperson's stab at this--please correct me if I've got it wrong.

    I think the term "weaponized" was a confusing term.

    In retrospect, it seems that some experts were using the term "weaponized" narrowly, to refer to a process of coating the prepared spores with something like lint which would make them bouncier and more likely to stay in the air for a longer period of time than ordinary spores. (Apparently only a few nation states knew how to do this, and the presence/absence of this treatment would draw the investigation either toward or away from the idea of nation-state-sponsored terrorism.)

    Others used the term more generally, to refer to the fineness of the milled spores, which were unusually small and "floaty."

    So in the end, no added linty coating=no "weaponization" per se, but the fineness and dispersability of the powder made the anthrax a pretty potent "weapon" in any casual sense.

    Then the term got bandied about, and the rest is history...

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    Unknown said...

    To be honest, I didn't understand what exactly led the FBI to suspect Steven Hatfill in the earlier years of the investigation? What are the commonly chosen topics to write about for a research paper?