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LONE WOLF - FBI K-9 Mysteries #1

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Biosecurity Incidents In Top U.S. Labs—What, Me Worry? Smallpox Edition

Smallpox is one of the few diseases that modern medical science has managed to eradicate. Believed to have emerged around 10,000 B.C., the disease first appeared in historical documents in the 15th century. The disease is estimated to have killed 300 to 500 million people in the 20th century alone, having a fatality rate of 20–60% in adults and 80% in children.

Smallpox is widely taught in university immunology classes as the world’s first vaccine. Edward Jenner noticed in 1798 that milkmaids who developed cowpox seemed to be immune to smallpox infection. In a stunningly unethical, off-the-cuff experiment, when a milkmaid came to him for treatment, he ran a thread through one of her pustules, coating it with pus. He then inoculated the eight year old son of his gardener by making a small cut and running the pus-coated thread through it. Shortly after, the boy became symptomatic for a mild case of cowpox. Several months later, Jenner took pox scabs from someone with small pox and similarly inoculated the boy a second time. The boy was immune to smallpox and remained healthy. This was the beginning of the vaccine revolution.

In 1967, the WHO mandated the eradication of smallpox, using newer, modern vaccines based on vaccinia virus, a virus related to both smallpox and cowpox. The last known case of smallpox occurred in 1977, and the WHO considered it eradicated in 1979.

This left the world with only laboratory strains of the virus. Following a breach of containment, resulting in the death of a lab worker in 1978, any labs with remaining virus either destroyed them or transferred them to safer labs. Currently, only the U.S. Centers for Disease Control and the Russian State Research Center for Virology and Biotechnology still retain samples. The argument has been made that all remaining virus should be destroyed, but the existing aliquots are retained in case any other stocks arise leading to a dangerous bioweapon in the wrong hands. At this point, decades after the final vaccinations, essentially everyone except military personnel (who continue to be inoculated with the vaccinia vaccine for out of country work) would be susceptible to a fresh onslaught of smallpox. But with only two stocks of the virus in the world, we’d like to believe that we’re safe.

So it was somewhat of a shock in July 2014, when the National Institutes of Health reported that six glass vials of freeze-dried smallpox stock had been found in a long forgotten box in the back of a cold storage room. I remember hearing the news and being stunned for several reasons—glass vials to store a biosafety level IV pathogen (of course, there was no sterile, disposable Nalgene polypropylene cryovials back then, but glass? So incredibly dangerous…), no security, and no inventory so no one even knew they were there. The stock was estimated to have been there since the 1950s, even though the building didn’t open until the 1960s, and from the 1970s on was used by the Food and Drug Administration. A further investigation reveals twelve boxes in total containing smallpox, dengue, influenza, Q fever, and rickettsia, all previously unknown to be stored there, and all with no security precautions (proper security precautions would involve a minimum of two locked doors between the pathogen and the public, detailed inventories, and full biosafety training of all personnel). The FDA immediately mandated a full review of all cold storage spaces to ensure no other pathogens were present.

The glass vials were immediately transferred to the CDC to undergo testing, where it was determined that two of the six vials contained viable virus. Had the tubes broken, the world could have seen a smallpox pandemic the likes of which it hadn’t seen for decades and for which we are entirely unprepared. Luckily for all of us, the vials remained intact. All the vials were destroyed following testing.

Ann and I are going to be taking some time off for summer holidays and to really concentrate on drafting LONE WOLF, book one of the new FBI K-9 Mysteries with Kensington Books. So we look forward to coming back fresh and with a lot of solid writing behind us in September. See you then!

Photo credit: Wikimedia Commons


Biosecurity Incidents In Top U.S. Labs—What, Me Worry? Influenza Edition

In the same report published by the CDC in July 2014 that discussed a laboratory incident concerning anthrax, an incident concerning influenza was also revealed. In August 2014, a full report detailed the cross-contamination of a non-pathogenic H9N2 strain of avian (bird) influenza with the highly pathogenic H1N1 strain (remember the 2009 flu pandemic? That’s the one…) which then sent it out to a U.S. Department of Agriculture lab that had no idea what they were dealing with. Fortunately, the Department of Agriculture lab is also a biosafety level III (BSL III) lab, so the samples were handled under BSL III containment procedures. As it turned out, no workers were infected with the pathogenic strain, a very lucky break as the story could have ended very differently.

As a scientist, the moment I heard this story, I knew exactly what had likely happened and it’s a big no-no in working with cells and viruses. Keep in mind, the error wasn’t identified until six months after the fact, so the worker couldn’t recall the day like it was the previous week, but reported working with then non-pathogenic H9N2 virus, decontaminating the biological cabinet and the working with the pathogenic H1N1 virus. However, of the 1.5 hours required to do all of this, key card access indicated the worker was only present for 51 minutes and that also included time to shower out of the facility and dress in street clothes. Clearly, the full protocol was not carried out. The scientists admitted that they were under pressure at the time to complete work for an upcoming WHO vaccine conference, and, that day in particular, the scientist in question was rushing to get to a lab meeting. Corners were clearly cut.

There are two scenarios that could have happened:

  1. Considering that each infection should take 30 minutes, it is possible the scientist did both infections in the biological cabinet at the same time. To put it plainly, this is NOT done. One of the first rules of tissue culture is that products are kept separate to ensure purity of the product and safety of the current and any future lab worker. I’d like to think this isn’t what happened.
  2. Instead of following protocol and working with the less pathogenic strain first and then the pathogenic strain, the scientist may have mixed up the order, working with the more pathogenic strain first and then not decontaminating afterward before moving onto the second, less virulent strain. Personally, I think this is what happened. Also, as PCR testing of the H9N2 strain doesn’t indicated H1N1 contamination, it’s likely they weren’t used concurrently.

One other concerning incident happened in association with this cross-contamination. When the receiving lab started to use the virus, chickens in the experiment started to unexpectedly die. Upon testing their virus, they determined that their H9N2 was contaminated with the deadly H1N1. When they informed the CDC, the lab team tested their stock of H9N2 to confirm that it was indeed contaminated with H1N1. But they did not report the incident at the time. It wasn’t until a second CDC team found atypical results with their stock of H9N2 virus that the original team reported the incident. At that time, all connected stocks of H9N2 were destroyed. Luckily all work done by the second team was conducted under BSL III containment, so there was no risk to any of the lab personnel.

For a group that is considered by most scientists as the gold standard, this incident combined with the anthrax incident is quite distressing. Scientists did not display good laboratory practices, training, communication skills, and in many cases, common sense. As a scientist myself, especially as one trained in BSL III procedures, many of these errors would simply not be acceptable or expected at our facility. Protocols and training are already in place to avoid this and a detailed incident reporting procedure is in place (and let me assure you, I’ve used it for BSL III incidents twice). After all this, I know I certainly look at the CDC differently, and I’m sure many other scientists do as well.

Next week we’ll be back with our last installment in this series. What happens when a virus that is nearly eradicated pops up in someone’s freezer? Come back next week and we’ll tell you all about it…

Photo credit: Wikimedia Commons


Biosecurity Incidents In Top U.S. Labs—What, Me Worry? Anthrax Edition

Today we’re continuing on with our series on laboratory biosafety and how it’s gone wrong lately in places you would think would be immune to such incidents. But it just goes to show human error can trump every precaution you put in place and that hopefully your people would follow to the letter. Especially when lives are on the line. But not so much, apparently…

In July of 2014, the Centers for Disease Control (CDC) held a press conference to discuss two different incidents that had taken place the previous month. We’re going to showcase one incident this week and one next week.

The first incident involved a CDC lab preparing extracts from anthrax, a biosafety level III (BSL III) pathogen. Their usual protocol involved chemically deactivating the pathogen for 24 hours, but they learned of a new protocol that only required a 10 minute deactivation. Now, this protocol was not for anthrax, but was instead for Brucella, another BSL III pathogen. However, they elected to try this new protocol on anthrax. This method also included a double check—after the pathogen is deactivated, some of it is plated onto agar plates and incubated for 48 hours to ensure there is no growth, and all the pathogen is dead. The CDC first attempted this protocol in June of 2014, sampling the extract to agar plates after 10 minutes, but leaving the rest for the full 24 hours.

However, due to a misunderstanding during a phone conversation and a lack of follow-up with the actual printed protocol, the scientist responsible only left the agar plates for 24 hours post deactivation, at which time, the plates were determined to have no growth. The scientist intended to autoclave the plates and discard them that day, however he could not open the autoclave door, so the plates were returned to the incubator. At this time, the anthrax incubated for 24 hours was distributed as deactivated pathogen to biosafety level II (BSL II) labs. Eight days later, the agar plates were removed from the incubator for disposal and, to their surprise, anthrax growth was observed. Scientists realized the 10 minute procedure was not sufficient to kill anthrax, but were unsure if the 24 hour procedure (from which anthrax had been sent out to BSL II labs and their lower level of containment) was sufficient. At that point, it had to be considered that they had a breach in biosafety containment and all the labs had to be completely decontaminated. Later tests showed that the 24 hour procedure was sufficient to kill most of the anthrax, but not all. As a result, while it was unlikely that infection would occur, it was not impossible.

Upon closer examination, there were a number of issues that led up to this incident:

  1. Use of unapproved techniques—there were several related to filtering of the extract, but this also included observing the plates for sterility at 24 hours instead of the required 48 hours.
  2. Transfer of material not confirmed to be inactive—based on the error made in point 1, this also involved a lack of written protocol of what was required to ensure that pathogens were truly deactivated.
  3. Use of pathogenic strains of anthrax to test out a new protocol when non-pathogenic strains would have revealed the same conclusion with none of the risk. This was an extremely unwise decision.
  4. Inadequate knowledge of peer-reviewed literature by both the laboratory scientist and his supervisor—papers already existed outlining that this method was not sufficient for absolute sterility of infectious anthrax.
  5. Lack of standard operating procedures to document pathogen deactivation.

Fortunately, no staff member ever presented with symptoms of anthrax. But the laboratory in question was closed pending a number of assessments, the establishment of new procedures, and until remedial action was taken with the staff involved. This was not a shining moment for the CDC, the institution we scientists like to consider the gold standard in biosafety. This is the group you call when something goes wrong and this is what their own people are doing?

In the last month, a new anthrax story surfaced concerning an Army lab at the Dugway Proving Ground in Utah. It creates anthrax test kits and sends them out to numerous laboratories for local testing. Contained in the kit is a radiation-killed sample of anthrax to use as a positive control for testing. However, in May, a Maryland biotech company identified that live anthrax was present in the samples. Upon closer inspection, it appears the samples were not sufficiently treated to kill all the spores. The test kits were packaged and sent out, many by regular FedEx delivery, to 69 labs around the United States, but also in Canada, Britain, Australia and South Korea. The CDC has confirmed that all kits shipped between 2004 and 2015 contained live anthrax.

Even more disturbing information surfaced this past week. It appears that from 2007 onwards, the lab was aware that their deactivation process (chemical deactivation at the time, and later irradiation) was not sufficient to kill all the anthrax, but they ignored the issue. At the time, federal regulators at the Office of the Inspector General were aware and recommended further investigation and potential enforcement action, however nothing was ever done. Furthermore, the information was never reported to Congress—the group responsible for oversight of this lab—so they were completely unaware of the situation. No laboratory workers have shown symptoms of anthrax; however 31 workers are receiving antibiotics as a precaution.

Next week, we’ll be returning the CDC as we discuss a serious mishap concerning a highly pathogenic strain of influenza.

Photo credit: Wikimedia Commons


A New Publishing Contract and Format For TWO PARTS BLOODY MURDER

We’re very happy to announce today that TWO PARTS BLOODY MURDER will be joining the ranks of DEAD, WITHOUT A STONE TO TELL IT and A FLAME IN THE WIND OF DEATH as part of Harlequin’s Worldwide Mysteries in mass market paperback format. We’ve been very pleased with our sales through Worldwide and are looking forward to our continuing partnership with them. Currently the projected publication date is spring 2016, but I’ll post the exact date on the website when we know it.

The deal was announced two days ago on Publisher’s Marketplace:

June 21, 2015 - TWO PARTS BLOODY MURDER by Jen J. Danna with Ann Vanderlaan

Jen Danna with Ann Vanderlaan's TWO PARTS BLOODY MURDER, the third novel in the Abbott and Lowell Forensic Mysteries, where a body discovered in a long-forgotten speakeasy proves to be ground zero for a cascade of murders through the decades, to Laura Barth at HQN Worldwide Mystery, for publication in spring 2016, by Nicole Resciniti at The Seymour Agency.


One of the fun aspects of working with Harlequin is that they always do their own back cover blurbs and cover art, so it’s always interesting to see their take on our series. As you can see above, they do an excellent job with series branding, so we can’t wait to see where they take TWO PARTS BLOODY MURDER. So we’ll be back with more on this release in the future as we have more information to share.


Biosecurity Incidents In Top U.S. Labs—Biosafety

This post may be a little off the beaten path for some of our blog readers, but it’s a story I’ve been watching over the past year or more. And when a new twist on the story hit the news in the last week, I thought it might be a good blog topic, even though it doesn’t have anything to do with writing or forensics. In this case, I’m looking at something near and dear to my day job—infectious diseases research.

In July, 2014, several alarming announcements came out of the U.S. Centers for Disease Control (CDC) involving potential and proven accidental exposures of lab personnel to extremely infectious pathogens. Then, that same month, the NIH announced the discovery of an even more dangerous pathogen found forgotten in FDA freezers. Just last month, the Pentagon admitted that one of its Army labs sent live pathogen out to as many as 69 different labs in a number of different countries. And now, this past week, it was revealed that same lab was secretly sanctioned for those actions eight years ago, but this information was hidden from Congress, which is responsible for oversight of the lab.

It’s enough to give a person nightmares about biological disasters.

Before we get into what went wrong with labs the world considers the gold standard in biosafety, let me give you a little bit about my background. I’ve worked at McMaster University in Hamilton, Ontario for nearly 25 years. I sat on the university’s Presidential Biosafety Committee for 5 years, making decisions on how to implement government standards for the safe use of pathogens, and how to rectify procedures that resulted in accident. I’ve worked nearly 25 years with biosafety level II pathogens (herpes simplex viruses I and II, vaccinia virus, Adenovirus, influenza, dengue virus and many others), and for over 15 years with biosafety level III pathogens (HIV and herpesvirus saimiri). Biosecurity has been my life for my entire adult working career.

What exactly does ‘biosafety level’ mean? The biosafety level of a pathogen is based on a number of factors: infectious dose (i.e. how much is required to contract the disease), mode of transmission (i.e. blood borne vs. air borne), host range (i.e. human vs. animal or both), the availability of effective treatment (i.e. pharmaceuticals), and the availability of preventative treatment (i.e. vaccines)

There are four internationally recognized biosafety levels for pathogens:

  • Biosafety level I—an organism unable or unlikely to cause disease in healthy individuals. May cause disease in immunocompromised individuals.
  • Biosafety level II—infectious organisms that cause disease, but are unlikely to make an individual seriously ill under normal circumstances. Effective treatment and preventive measures are available, and the risk of spread is limited. Example: herpes viruses, including those that cause mononucleosis, chicken pox and roseola.
  • Biosafety level III—infectious organisms that cause serious disease, but do not spread by casual contact. Organisms that cause diseases treatable by antimicrobial or antiparasitic agents. Example: HIV, anthrax.
  • Biosafety level IV pathogens—infectious organisms that cause very serious disease, often untreatable and leading to death, and are spread by casual contact. Example: Ebola virus.

Different containment levels are required to study pathogens at different levels.

  • Biosafety level I pathogens, like many strains of non-infectious E. coli, can be used on the open bench in the main lab.
  • Biosafety level II pathogens must be used within a separate level II room in a biological safety cabinet. Air is drawn into the cabinet away from the user and only exhausts into the room after HEPA filtration, thereby keeping any infectious particles trapped inside the cabinet. The user only interacts with the pathogen after donning a lab coat and gloves and by remote access i.e. using a pipettor to actually contact/transfer the pathogen.
  • Biosafety level III pathogens are used in an isolated, approved access-only laboratory with negative pressure similar to a biosafety cabinet—air is drawn in from outside the laboratory and only exhausts from a laboratory through giant HEPA filters in the ceiling. Workers inside the facility must wear rear-closing gowns and double gloves at all times. In the case of airborne pathogens, workers must also wear respirator hoods with a powered air purifier worn on the belt. A safety shower must be located in the laboratory in case of spills and pathogen appropriate disinfectants must be available at all times. In case of incident, detailed reporting and follow-up is mandatory.
  • Level IV pathogens can only be used in biosafety level III style labs situated in an isolated building. Work will involve all the requirements of biosafety level III work, but over and above that, the use of a positive pressure personnel suit (which pushes air out of the suit and away from the user) is mandatory at all times. Multiple showers (both in-suit and out), a vacuum room, and a UV room are required for clean exit from the facility.

Specialized training for each specific biosafety level is required by all personnel, often with annual updates and review. As you might imagine, the complexity of the training increases in orders of magnitude as the biosafety level increases.

Another aspect that is crucial to biosafety work, especially at upper levels, is tracking—where pathogen is stored, how it is treated, when small volumes are removed, how they are used, and how they are destroyed. There are strict guidelines concerning any transfers outside the laboratory as well—how it can be removed and who is allowed to receive it based on their facilities and training.

So now that you understand some of how this work is done, next week we’re doing to look at what went wrong (when it absolutely shouldn’t have) and what’s being done to safeguard workers and the public. See you then!

Photo credit: Wikimedia Commons