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The threat of bioterrorism: Identifying the unknown

No. 02

April 2012


The threat of bioterrorism: Identifying the unknown Janet Martha Blatny and Per Leines Lausund, Norwegian Defence Research Establishment (FFI)

Biological threats may pose a risk to the Armed Forces as well as to civilians. It is therefore critical to understand and evaluate biological threat agents and their potential areas of misuse. How can we be better prepared to manage bioterrorism and its consequences? And how we can reduce the risk of such incidents in the first place?

The Norwegian Defence Research Establishment (FFI) has been addressing biological threat challenges since 1998. Under contract with the Norwegian Ministry of Defence, it provides advice on preparedness and response for such incidents by analysing technical challenges and potential misuse of biological threat agents. It possesses amongst other things a unique national laboratory facility for confirmed identification of samples containing chemical, biological and radiological (CBR) agents either alone or in a mixture. The laboratory is available to the Armed Forces, as well as to civilian authorities and institutes. The objective is how to better identify the unknown. Fear as a weapon

One of the first steps in managing a biological crisis is to ascertain whether a biological incident is a real threat, a hoax or a natural outbreak. Therefore, FFI has, in compliance with NATO concepts and requirements, established a wide set of methods and guidelines regarding sampling, preparation, confirmed identification and microbial forensic analysis of biological threat agents with emphasis on environmental samples. Together with risk- and vulnerability assessments on pandemic flu, the threat of bioterrorism constitutes a main focus of infectious diseases and public health


The threat of bioterrorism: Identifying the unknown

preparedness work outside hospitals. Experts differ in their threat assessments of bioterrorism. Some claim the threat is dire and imminent, others the opposite. Moreover, it is widely agreed that fear is a major impact of bioterrorism, making it an attractive tool for terrorists. Biological agents are defined as microorganisms, or toxins, that are able to cause disease, damage or death to humans, animals, plants or materials. The terms “biological warfare agent” or “biological weapon” are also often associated with these types of agents. With respect to bioterrorism, the term “biological threat agent” should be used, as a biological weapon refers to a proven and tested delivery system containing an agent. International accords

The Geneva Protocol was issued in 1925 to ban as weapons of war the use of asphyxiating, poisonous, or other gases. These substances had been used during World War 1 causing close to one million casualties. In comparison, the Spanish flu caused between 15 and 40 million casualties in 1918-1919. The Biological and Toxin Weapons Convention (BTWC), reaffirming the Geneva Convention, strictly prohibits the production, development, stockpiling and use of biological weapons and was opened for

signature in 1972 and effective as of 1975 ( It also bans "weapons, equipment or means of delivery designed to use such agents or toxins for hostile purposes or in armed conflict.” However, BTWC has no verification regime, and does not allow the intrusive inspections which the Chemical Weapons Convention (CWC) permits. As of December 2011, 165 nations have ratified the Convention. Norway ratified the BTWC in 1973 and the Norwegian Ministry of Foreign Affairs, assisted by FFI when needed, plays an active role in its commitments to that end. In addition, several countries have put in place comprehensive safeguards to prevent misuse of biological threat agents (one example is The “golden age” of biological weapons (BW) stretched from the period between the two world wars and well into the 1970s. Several countries, including the U.S. and the U.S.S.R. possessed comprehensive programmes. Biological weapons were effective as tested population killers with well-documented effects. Agents causing anthrax, plague, smallpox and botulinum toxin were regarded as the most suitable and feared in the BW programmes by the U.S., the U.S.S.R., Germany, Japan, the U.K., and Canada regarding pathogenicity and virulence.


B. anthracis was also first choice when Iraq started developing a BW capability in the late 1980s. Iraq had produced 19,000 litres of botulism toxin, 8,000 litres of anthrax and 2,000 litres of aflatoxins during their BW programme, discovered by the United Nations Special Commission (UNSCOM).

between 1900 and 2003 where biological threat agents were used. It showed that 73 percent of the non-state sponsored incidents were perpetrated by individuals with scientific or medical training.

The unintentional release of a small amount of anthrax spores from one of the BW research facilities in Yekaterinburg in Sverdlovsk, Russia in 1979 unwittingly demonstrated the effectiveness of infection by inhalation, while the postal anthrax case in the U.S. in 2001 exposed a nation’s vulnerability to a small-scale dispersion of anthrax spores (bacterium Bacillus anthracis).

A “successful” use of biological threat agents (to cause infection) depends on the perpetrators knowledge, skills and abilities. Also, counter-proliferation and -security efforts undertaken by most nations are important aspects in denying non-state actors’ access to biological threat agents. For instance, in 1994-95, the Japanese religious cult Aum Shinrikyo attempted to disperse B. anthracis spores in Japanese cities, but despite massive investments they failed due to the unintended use of a non-virulent bacterial strain.

A previous review addressing the majority of incidents between 1960 and 1999 using biological material in order to kill, incapacitate, or threaten showed thatricin, HIVinfected blood, and food contaminants (for example Salmonella species and Shigella species were frequently used). A separate study investigated 29 confirmed incidents

The biological threat can never be ignored as long as it remains possible to misuse and deliberately disseminate disease causing agents. This because knowledge of how to modify the infective properties of for instance influenza A/H5N1 viruses to be transmitted efficiently between mammals, still maintaining high pathogenicity, does in fact exist

Bioterrorist attacks

NATO trial. Scientists at FFI had to identify parcels containing an unknown content of CBR materials during a NATO trial at FFI, 2010.

The threat of bioterrorism: Identifying the unknown



Fortunately, serious technical obstacles must be overcome, for instance to produce a gram of weaponised B. anthracis spores. However, once that has been accomplished, it is easy to manufacture larger quantities. One of the ongoing projects at FFI aims precisely to identify and understand such obstacles in the production of biological threat agents, to assist in preventive actions regarding the misuse of such dual-use technology and equipment. Synthetic biology

Modern strategies of DNA-synthesis have made it possible in principle to re-create all known viruses independent of natural templates. This is known as synthetic biology. As many of the select agents have been sequenced, such as Ebola virus, 1918 Spanish Flu virus, smallpox and SARS, it is no longer an extensive challenge to (re) create these organisms. Only the sophisticated knowledge of a multidisciplinary field of experts is needed. Several viruses have been synthesised, including the polio virus in 2002, the H1N1 1918 Spanish Flu in 2005, the West Nile virus in 2010, and SARS in 2008. Viruses have smaller genomes than bacteria (< 30 kilobases, kb), but it has been demonstrated that new cells of Mycoplasma mycoides may be created in vitro (1, 08 megabase, Mb genome). Nucleotide sequences need to be synthesised to (re)create microorganisms, and the purchase of such synthetic genes from commercial institutions is now routine in scientific communities. Unfortunately, not all commercial institutions check and compare ordered nucleotide sequences for possible similarities to sequences found in pathogenic microorganisms. Such comparison schemes are an important step in reducing the threat of recreating/creating pathogenic organisms for use by bioterrorists. Recently, scientists have genetically altered the H5N1 avian influenza genome, resulting in an infective virus capable of airborne passage between ferrets. After passing the genetically modified H5N1 virus between ferrets a highly pathogenic virus was obtained with only five mutations in two genes, and with the potential to be transmissible between humans. This raised great concerns about the consequences if the virus were to escape the laboratory or fall into the hands of bioterrorists. Some even question whether such results should be published at all. For instance, the US National Science


The threat of bioterrorism: Identifying the unknown

Advisory Board for Biosecurity (NSABB) decided to restrict some of their research results from being published due to the potential risks of misuse.

In the chamber. Janet Martha Blatny and Jaran Strand Olsen at work in FFIs aerosol chamber. The chamber is used for characterizing bioaerosols, which are microscopic airborne particles, and for testing and evaluating equipment for collection, detection and identification of bioaerosols.

Synthetic biology causes concern as the technology may be used legitimately for

Today, technology needed for genetic engineering is available for purchase, also second-hand. This allows for so-called “do it yourself”-biology. human betterments, but also misused by bioterrorists. This underscores the need for awareness of possible illegitimate use of knowledge and laboratory facilities to develop threat microorganisms that may have increased pathogenicity. Such awareness should be part of any students’ training, also emphasizing the BTWC and individual responsibilities as steps to reduce the risk. “Do it yourself”-biology

Today, technology needed for genetic engineering is available for purchase, also second-hand. This allows for the construction of synthetic genes and genetically modified cells in a hobbyist’s basement or garage; so-called “do it yourself”-biology. Such developments suggest that biological threat agents soon could be designed by grass-roots biohackers, not just by highlyskilled scientists. FFI is carefully monitoring international scientific progress looking for new routes in the (re)crea-


tion and production of possible new and (re)emerging biological threats. Only by identifying such routes reliable advice on countermeasures can be provided. Is it possible to distinguish between natural outbreaks on one hand, and outbreaks caused by the deliberate use of biological threat agents on the other? In principle, the result of bioterrorism could be the same as the result of a natural outbreak. However, from the point of view of a terrorist, a successful attack should trigger as many simultaneous effects as possible among the exposed individuals. Besides demonstrating that it is a deliberate event and maximising injury, this would also minimise the ability of health services to respond appropriately. The strain of the microorganism identified in an incident may also lead to suspicion as to the cause of the outbreak, especially if it is unusual in that geographic area. Physicians need to be on the alert and include such unusual agents in their diagnostic panels. Biological defence capability

Biological threat agents may be difficult

to detect and identify reliably and rapidly from a public health and military perspective. Internationally, biodefence capability gaps are continuously identified and great efforts are invested in improvements. The biodefence area covers some aspects, but an essential part is the ability to respond and react quickly. NATO, the European Defence Agency (EDA) and the European Commission (EC) have each established concepts describing detection, identification and monitoring (DIM) for chemical, biological, radiological and nuclear agents (CBRN). FFI plays an active role, including as coordinator, of several projects within NATO, EC and EDA, addressing these challenges. These groups are important forums for exchange of information as well as for developing harmonised solutions against possible CBRN challenges. There are several methods in use to identify biological threat agents, but no single approach to obtain confirmed identification exists. Several methods are needed, and in vivo studies may be required for unambiguous identification. Identification of strains causing outbreaks is often time consuming due to epidemiological studies,

sampling, cultivation techniques and the problems of new or rare strains/serotypes. One example is the German outbreak of the enterohaemorrhagic Escherichia coli EHEC 0104:H4 in May 2011, which stemmed from fenugreek seeds imported from Egypt and used to produce sprouts. FFI has also established various methods for genetic fingerprinting (genotyping), particularly useful for microbial forensics. Identification of Salmonella typhimurium as the causing strain in the deliberate outbreak of salmonellosis by the Rajneeshee Cult in 1984 (see fact box page X) took four days, but it took more than a year to identify and confirm that only a single strain of S. typhimurium had been used - after the confession of one of the cult members. Many bacterial threat agents occur naturally, and others may be closely related to other bacteria found in the environment. Thus, early identification and analysis must distinguish between naturally occurring outbreaks, background flora and the release of biological threat agents. Early detection and rapid identification are both critical to minimise casualties and affect a timely and efficient response and containment to limit the outbreak.

ď&#x20AC;ľResearch. Marius Dybwad is a PhD-scholar at FFI. His main research includes bioaerosol characterization and its impact on biological detection.

The threat of bioterrorism: Identifying the unknown



BIOWEAPONS TIMELINE  1346 During the Middle Ages, the Tartars are said to have catapulted bodies of

plague victims over the walls of Kaffa in an attempt to initiate an epidemic upon the residents.

 1763 Blankets from smallpox patients were given to the Native American tribes in

order to transfer the disease and influence the outcome of the ongoing conflict.

 1914-1918 The Germans used biological agents for sabotage such as infecting

animal feed and horses intended for export.

 1925 The Geneva protocol bans the use of bioweapons.  1937-1945 In response to suspected bioweapon (BW) development in Nazi

Germany, the US, UK, and Canada initiated a BW development program in 1941 resulting in the weaponisation of anthrax, brucellosis, and botulism toxin. Japan also had a comprehensive program, mainly on plague.

 1942-1969 Center for U.S. military BW research developed and tested biological and

chemical weapons at the Dugway Proving Grounds in Utah. Research carried out in the U.K. during World War II left Gruinard Island in Scotland contaminated with anthrax for the next 48 years. In the USSR, a biological weapons program continued until the dissolution of the union.

 1972 The Biological and Toxin Weapon Convention (BTW) banned the use,

possession and development of chemical and biological weapons.

 1978 KGB-agents killed the Bulgarian dissident Georgi Markov by stabbing and

injecting him with ricin. In the Soviet Union, toxin from Ricinus communis, was redefined as an assassination tool at the time.

 1979 An Anthrax epidemic broke out in humans in the city of Sverdlovsk in the

former USSR. The reason was an accidental release from the BW-research facility in Sverdlovsk, resulting e.g. in 66 human deaths.

 1984 The Rajneesh Cult deliberately released Salmonella typherium at salad bars

and supermarkets in Oregon, USA, causing an outbreak of salmonellosis where 751 people fell ill.

 1985 The inception of Iraq's biological weapons program, which embraced a

comprehensive range of agents and munitions. Agents under Iraq's biological weapons program included lethal agents, e.g. anthrax, botulinum toxin, aflatoxin and ricin, and incapacitating agents, for example some mycotoxins, haemorrhagic conjunctivitis virus and rotavirus.

 1995 Members of the Aum Shunrikyo sect released sarin on several lines of the

Tokyo Metro, killing thirteen people, severely injuring fifty and causing temporary vision problems for nearly a thousand others.

 1996 On 28 August, in Dallas, Texas, USA, 12 laboratory workers at St Paul’s Medical

Centre became ill after eating muffins and doughnuts in their cafeteria. Apparently, the food had been intentionally contaminated with Shigella dysenteria type 2 by a disaffected worker.

 2001 In the aftermath of 9/11, the dissemination of anthrax spores by letters and

the postal processing and distribution centers in the United States resulted in 22 cases of anthrax, of which five of the inhalation cases were fatal, and more than 2000 treated with antibiotics in one of the postal facilities alone.

 2011 North Georgia men arrested, charged in plots to purchase explosives,

silencer and to manufacture a biological toxin for use in attacks against U.S. citizens and government personnel and officials.


The threat of bioterrorism: Identifying the unknown

Studying natural outbreaks is useful to learn how to respond to bioterrorist incidents, but the analysis of microbial evidence from a bioterrorism incident is also crucial in identifying the perpetrator, and must be factored into the investigation process. An efficient biological preparedness and response requires close collaboration between the human and veterinary medical fields, practitioners, public health, first responders, police and researchers, as well as policy- and decision makers within ministries and directorates. Early warning systems

One step in reducing the impact of outbreaks and diseases is to provide early warning systems (syndrome based systems) to strengthen and complement continent-wide disease surveillance systems. In Norway, physicians are required by law to notify cases of certain infectious diseases to the Norwegian Surveillance System for Communicable Diseases (MSIS) at the Norwegian Institute of Public Health. Nevertheless, an early identification of the biological threat agent before the clinical symptoms appear will make it possible to reduce the infection rate, casualties and the people exposed to the threat. This will also allow for adequate medical and protection countermeasures to be initiated early to increase their effect. There is also a need to develop technology and systems that aim to detect and identify biological threat agents on the scene of contamination following exposure. For military operations, it is critical to be warned as early as possible to initiate proper countermeasures. In the event of a biological crisis, extensive investigation is required. This often implies analysis of many samples from the environment at reach back laboratories, as well as samples from exposed individuals to be analysed at hospitals. Thus, a well-defined biological preparedness system requires an efficient and alert medical service system and emergency support teams addressing the incident after it has occurred as well as necessary capabilities alerting authorities prior to such incidents. Furthermore, multi-disciplinary training (table-top and operational) based on realistic scenarios is needed, ensuring a well-communicated reporting and response system among those involved.


ď&#x20AC;ľBacterial growth. Airborne bacteria grown on an agar plate in the FFI lab.

An active contributor

FFI addresses the biological threat by analysing the technical potential of misuse of biological agents in bioterrorist acts, biocrime or other events where such agents are deliberately used. This work contributes towards increasing the understanding of how to reduce the risk of bioterrorism and how to improve preparedness. Moreover, it constitutes the basis for biological threat assessments

FACTS CDC AGENTS LIST The Centre for Disease Control (CDC) in the U.S. has established a select agents list, classified in the categories A, B and C, including certain agents which could be exploited by terrorists. Category A includes high priority agents that i) pose a risk to the national security since they may easily be disseminated, ii) are transmitted from person to person, iii) may result in high mortality rates, and iv) may cause public panic and require special health preparedness. These agents include Bacillus anthracis (anthrax), Francisella tularensis (tularemia), Yersinia pestis (plague), Variola major (smallpox), viruses causing viral hemorrhagic fevers and botulinum toxin (botulism). Category B agents consist of biological threat agents that are moderately disseminated and result in low mortality rates. This category includes Coxiella burnetti (Q-fever), Brucella spp. (brucellosis), Burkholderia spp. (glanders, melioidiois), viruses causing viral encephalitis, Rickettsia prowazekii, (typhus fever), and waterborne and food safety threats such as Vibrio cholera (cholera), Shigella and Salmonella species, respectively, in addition to the toxins ricin, Staphylococcus enterotoxin B (SEB) and epsilon toxin of Clostridium perfringens. Category C agents include agents of emerging pathogens that could be engineered for mass dissemination due to their availability, ease of production and dissemination, as well as their potential for high morbidity and mortality rates and major health impact. Such agents may be exemplified by Nipah virus and hantavirus.

outlined by the responsible authorities; the Norwegian Police Security Service and the Norwegian Intelligence Service. The ongoing work at FFI is performed in close collaboration with Ministries, directorates and responsible authorities involved in biological preparedness and response. Ongoing scientific studies at FFI include detailed characterisation of biological aerosols (aerosols are dispersion of solid and liquid particles suspended in gas or air), agent-fate analysis, evaluation of technological detection systems, DIM capabilities, and developing mathematical models simulating biological agent dispersion and dissemination. We participate with our research in various programme activities within NATO, EDA, EU and NORDEFCO (Nordic defence Cooperation). The institute has bilateral agreements with equivalent institutes in Europe, US and Canada, and a trilateral agreement with the UK and the Netherlands (the ANNCP collaboration). FFI takes part in national biological preparedness, and assists the Armed Forces and the Ministry of Defence by enhancing CBRN response and capabilities. But access to expertise at FFI is not limited to the Armed Forces and the Ministry of Defence. It is also available for civilian authorities and stake-holders addressing bioterrorism and biological preparedness and response. Our objective is to reduce the risk of exposure to such agents and provide support and rapid response in the event of a biological crisis.

The threat of bioterrorism: Identifying the unknown


AUTHORS JANET MARTHA BLATNY is a chief scientist and project manager at FFI, where she works mainly with biological threats. Blatny has a Master of Science in Biotechnology Engineering and a PhD, both from the Norwegian University of Science and Technology (NTNU). She is also an active participant in work at NATO, the European Defence Agency, and the European Commission.

PER LEINES LAUSUND , is a doctor of veterinary medicine and lieutenant colonel. He works as a scientist at FFI and for the Defence staff. He graduated in 1982 from the Norwegian School of Veterinary Science with a master in public health. Lausund has held civilian and military positions, and was a section chief at the Norwegian Army Medical School. His work is mostly related to epidemiology and biological agents, and he has done national and multilateral work on bioterrorism and biological warfare. Lausund is represented in several NATO groups.


Norwegian Defence Research Establishment Instituttveien 20 P.O.Box 25 N-2027 Kjeller FFI HORTEN Karljohansvern P.O.Box 115 N-3191 Horten Telephone: +47 63 80 70 00 Telefax: +47 63 80 71 15 ‒ Photos: FFI Print: 07 Gruppen Text, photo og design: FFI ISSN 1503-4399 FFI is member of "Grønn stat". FFI FOCUS is printed on recycled papir.

Wimmer, E, and Paul, A.V. 2011 Synthetic Poliovirus and Other Designer Viruses: What Have We Learned from Them? Annual Review of Microbiology, 65: 583-609

tentional Epidemics (Medical Aspects of Biological Warfare)

Gibson, D. G., et al (2010) Creation of a bacterial cell controlled by a chemically synthesized genome. Science express 10.1126.

Török, T. et al, Tauxe, R., Wise, R., Livengood, J., and Sokolow, R. (1997) A large community outbreak of salmonellosis caused by intentional contamination of restaurant salad bars JAMA, 278:389-395

Smart J. K., 1997, in: Textbook of Military Medicine, Medical Aspects of Chemical and Biological Warfare (F. R. Sidell, E. T. Takafuji, and D. R. Franz, eds.), Office of the Surgeon General, Washington D.C., pp. 9 9ng NATO handbook for sampling and identification of biological, chemical and radiological agents (SIBCRA), AEP66, Dec 2009 Martin, W. et al, James W. Martin, MD, FACP*; George W. Christopher, MD, FACP†; and Edward M. Eitzsen, JR, MD, MPH‡ (2007) History of biological weapons: From poisoned darts to in-

Myhre, E. B. (2005) Biological weapons inspections - the Iraq experience

Dybwad et al (2012). Dybwad, M., Granum, P.E., Bruheim, P., Blatny, J.M. (2012) Microbiological Characterization Of The Bioaerosol Environment At An Underground Subway Station. Appl Environ Microbiol. 78(6):1917 Tjarnhage, T. et al, Jonnson, P., Blatny, J.M. Skogan G., Humppi, T. (2011) Detection of Airborne Biological Agents. NORDEFCO report FOI-R-3267-SE. Aas. P. et al, Blatny, J.M., Hegsvold, E. H., Jargren, E. (2011) BIOEDEP – et EDA program for utvikling av utstyr for de-

FFI FOCUS is FFIs publication for defence related subjects. These publications present themes from all of FFIs research. Contact for more information.

teksjon, identifikasjon og overvåking av biologiske trusselstoffer. FFI report 00624. Blatny, J.M. et al, Reif, B.A.P., Skogan G., Andreassen, Ø., Høiby, E.A., Ask, E., Waagen, V., Aanonsen, D., Aaberge, I.S., and Caugant, D.A. (2008) Tracking Airborne Legionella spp. and Legionella pneumophila at a biological treatment plant. Environmental Science and Technology, 42: 7360-7367. Meselson, M. et al, Guillemin, J., HughJones, M., Langmuir, A., Popova, V., Shelokov, A., and Yampolskaya, V. 1994. The Sverdlovsk anthrax outbreak of 1979. Science, 266:1202-1208. Melin, L. 2000. Terrorism och kriminalitet. FOI report 1551-864. (in Swedish) /482153a. Berns et al., 2012. Nature Feb 9, 342:482: 153-154

FFI-FOKUS nr 2 2012  

The threat of bioterrorism: Identifying the unknown. Biological threats may pose a risk to the Armed Forces as well as to civilians. It is t...

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