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

Trials and Tribulations of Starting a New Series

It’s been a while since I did an update from the writing trenches. As I’m now back into the writing of the first book of the FBI K-9 Mysteries, LONE WOLF, I thought now would be a good time.

After recently completing the fifth book in the Abbott and Lowell series, I’m finding it interesting (also, just a little frustrating) how difficult writing this new book is. Stepping into a Matt and Leigh book is like meeting up with old friends and, in a lot of ways, the characters write themselves. The setting is established, minor characters get the opportunity to grow, and the science is familiar and comfortable. It’s up to us to find interesting cases with a twist, but a lot of the backstory is already in place. There’s no mental gymnastics to get rolling, not at this point, as the time when they were new and shiny seems a long way back (2009 to be exact).

So here I am—new location, new law enforcement setting, new characters, forensics I’m not familiar with, technology I need to get up to speed on, new research contacts to make, and a whole field to become familiar with (dogs) so I don’t drive Ann-the-dog-expert completely insane correcting my every small error (let’s try to cut it down to the big errors alone because there’s no way I’m going to get this 100% right). As a result, when I sit down at the keyboard, it’s hard to get in the zone and get writing as fast as I’d like. Yes, we have wiggle room in the schedule for me to stumble a bit now, but I hate doing that. I like having a schedule and sticking to it. On the bright side, I look at what I’m writing and I can see it’s not garbage, so what is going down is working, it’s just a little like pulling teeth at this point. But we’re just past the 20% mark now and only slightly behind my self-imposed schedule with an end of August deadline, so all is not lost.

It doesn’t help that the day job has never been crazier; in fact, I suspect this has a lot to do with the writing not flowing so easily. My extremely competent and bench-savvy co-worker was recruited out of the lab to work in a downtown Toronto biotech company. I couldn’t be happier for him and he well deserves the position, but man, oh man, I want him back! In the meantime, we’re wrapping up multiple projects (one, a five year international clinical study which is HUGE) and, with the help of three fantastic students—one of whom is my own daughter—we’re going to survive this summer and get the work done. All while completing a draft of the first book in a brand new series and essentially not seeing more than 2 days of vacation until the fall.

It’s at moments like this that I remember Nora Roberts’ famous quote: “Every time I hear writers talk about ‘the muse,’ I just want to bitch-slap them. It’s a job. Do your job.” I think I need that on a poster to hang near my laptop. Nora, you’ve got it absolutely right. Butt in chair, hands on keyboard, do the job, and this book will get written on time!

Photo credit: Jeroen Bennink

Forensic Case Files: The Oldest Known Murder

There’s a lot we’ve learned about our distant ancestors, the many species that dead-ended or contributed to the evolution of Homo sapiens. But there’s a lot that skeletal remains and fossil records don’t contain—direct evidence of interpersonal relationships, for example. This past week, a team of Spanish, American, French and Chinese scientists was able to shine a light on our long past ancestors when they revealed the oldest murder victim yet on record dating from 430,000 years ago (the original open access paper can be found here).

If you’re like me, that’s such a long time ago that it’s almost impossible to wrap your head around it. 430,000 years ago, we didn’t exist as a species. That was during the Pleistocene epoch, a geological age that lasted from 2,588,000 to 11,700 years ago and included the last of our recent ice ages. The particular species in question, Homo heidelbergensis, is from the same genus as us (Homo) but differs at the species level (heidelbergensis vs. sapiens; although to be more exact, modern man is considered a more specific subspecies, Homo sapiens sapiens). Scientific opinion on our relatedness to Homo heidelbergensis differs, with some scientists considering us to have evolved from them, while other theories place them on another branch of the evolutionary tree altogether. Either way, these were people who roamed northern Spain approximately 230,000 years before we first appeared on the African savanna.

Researchers discovered a mass grave when they were excavating a cave named Sima de los Huesos, Spanish for the ‘Pit of the Bones’, in the Atapuerca mountains in the north of Spain. This mass grave represents the earliest known funerary rights of early man.

The cave is located at the bottom of a 43-foot vertical shaft and contained a mixture of animal bones and the remains of 28 individuals. At first scientists questioned if some of the remains in the cave were the result of a catastrophic fall; some may have been, but overall antemortem vs. post-mortem bone damage of the remains contradicts this theory. However, the presence of one individual leads them to believe that many of them were deposited there after death by their fellow hominins.

Cranium 17 (pictured above), belonging to a young adult, was found within that cavern. Exhibiting two distinctive penetrating and fatal wounds above the left eye, it is clear that this individual did not die of natural causes. Due to the placement and pairing of the wounds, researchers theorize that a face-to-face encounter of ‘interpersonal violence’ occurred leading to the death of one of the two combatants. They also theorize that due to duplicate weapon strikes, either of which would have been fatal, the intention was clearly to kill and the second strike was simply a safeguard in case the first was insufficient to cause death.

Researchers have studied the wounds both microscopically and via CT scan. Measuring nearly an inch across, both wounds appear to have come from the same weapon as a similar notch appears in both defects (the black arrow marking T1 and T2 below). One aspect of the CT scan that struck me is that the weapon created a similar oblique fracture angle on the underside of the skull (see the red box in the scan below) that forensic anthropologists note when describing bullet injuries to the skull.

It is possible that the injury may have been accidental, but the mechanics of such a front-facing injury imply that it was likely intentional. The theory is supported by the presence of two identical weapon strikes. You might accidentally hit a person or be hit by something once, but twice is likely an intentional act. Furthermore, a wound on the left side of the body in a front-facing attack implies a right-handed attacker using a standard tool. In fact, the authors suggest a wooden or stone-tipped spear or a stone axe handle might have been the weapon of choice.

While interpersonal violence has been well documented since Neolithic times (10,200 to 2,000 B.C.), it has always been assumed to exist in earlier man’s interpersonal relationships. However, this is the first documented case of direct trauma responsible for an intentional death in one of modern man’s earliest ancestors.

Photo credit: Javier Trueba