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PubMed2006

Fungi as bioweapons.

Fungi cause disease directly by infection or indirectly through mycotoxins. Fungi that are used as weapons might be targeted against humans, livestock, or crops. Humans and animals encounter fungi and mycotoxins through inhalation, ingestion, and contact with skin and mucous membranes. Effective fungal bioweapons would require the ability to cause significant destruction and a means of delivery to target populations or farms. Effective counter measures against fungal bioweapons would be able to prevent or treat this damage. This article describes several potential biological warfare or bioterrorism fungal species and mycotoxins in regard to their biology, epidemiology, potential for weaponization, and the clinical features, prevention, and treatment of the diseases that they cause.

Biological Warfare↗

PubMed2005

Biodefense research: an emerging conundrum.

Biodefense research creates a conundrum for the scientific community--what are the permissible limits of biodefense research and how can knowledge in the life sciences be protected against misuse? As biodefense research expands to counter the threat of bioterrorism, so does suspicion and the need for guidance to respond to these concerns.

Biological Warfare↗

PubMed2004

Anthrax.

Anthrax is an ancient disease associated with the plagues in biblical Egypt and modern bioterrorism. Three clinical syndromes result from exposure to anthrax spores: cutaneous,inhalational, and gastrointestinal. Cutaneous anthrax is the most common naturally occurring syndrome; inhalational anthrax is most likely to result from airborne release of spores. Prophylactic and early treatment can improve the mortality from inhalational anthrax. A vaccine is available, but has many limitations. New vaccines are currently being developed.

Animals↗

PubMed2004

Plague.

Plague is a disease that has been present for thousands of years and described since the earliest medical accounts. It occurs today worldwide, and may present in a variety of clinical forms. Bubonic disease, pneumonic plague, and septicemic plague are seen in addition to a number of other less common manifestations. As an agent of bioterrorism,Yersinia pestis could pose an extreme threat if released in the appropriate form and in the appropriate environment. Presumptive diagnosis may be made with readily available techniques, but laboratory handling of specimens requires special care. When there is a strong suspicion of plague, treatment should be instituted immediately, as delaying therapy will result in increased morbidity and mortality.

Anti-Bacterial Agents↗

PubMed2006

Evaluation of the rapid analyte measurement platform (RAMP) for the detection of Bacillus anthracis at a crime scene.

The aim of this study was to evaluate the accuracy and reliability of the rapid analyte measurement platform (RAMP) for presumptive identification of Bacillus anthracis spores. Test samples consisted of serial dilutions of spore preparations of several Bacillus species, including B. anthracis, which were tested, using the RAMP Anthrax test cartridge, according to the manufacturer's instructions. The fluorescence labelled antibody-antigen complexes were detected in the portable reader after 15 min following sample addition. Dilutions of common environmental and household powders were also tested to identify possible false positive results. B. anthracis spores were identified reliably in test samples containing more than 6000 spores. The test kits were highly specific, showing no cross reactivity with other Bacillus species or any environmental powders tested. The RAMP system for detection of B. anthracis spores, from environmental samples, showed consistent results under a variety of analytical conditions, enabling the trained user to provide a rapid, accurate preliminary risk assessment of a suspected bioterrorism incident.

Bacillus anthracis↗

PubMed2006

Vaccinia immune globulin: current policies, preparedness, and product safety and efficacy.

In 1980 the World Health Organization declared that smallpox was eradicated from the world, and routine smallpox vaccination was discontinued. Nevertheless, samples of the smallpox virus (variola virus) were retained for research purposes, not least because of fears that terrorist groups or rogue states might also have kept samples in order to develop a bioweapon. Variola virus represents an effective bioweapon because it is associated with high morbidity and mortality and is highly contagious. Since September 11, 2001, countries around the world have begun to develop policies and preparedness programs to deal with a bioterror attack, including stockpiling of smallpox vaccine. Smallpox vaccine itself may be associated with a number of serious adverse events, which can often be managed with vaccinia immune globulin (VIG). VIG may also be needed as prophylaxis in patients for whom pre-exposure smallpox vaccine is contraindicated (such as those with eczema or pregnant women), although it is currently not licensed in these cases. Two intravenous formulations of VIG (VIGIV Cangene and VIGIV Dynport) have been licensed by the FDA for the management of patients with progressive vaccinia, eczema vaccinatum, severe generalized vaccinia, and extensive body surface involvement or periocular implantation following inadvertent inoculation.

Bioterrorism↗

PubMed2005

Performance of a rapid dermatology referral system during the anthrax outbreak.

The bioterrorism-related anthrax outbreak generated unanticipated demand for dermatologic services. In this study we sought to perform rapid, efficient, cost-effective evaluation of patients suspected of having cutaneous anthrax. During the outbreak, we developed an anthrax evaluation system featuring clinical field examination by nondermatologist physicians, followed by rapid referral of selected high-risk patients to a centralized dermatology center. We excluded anthrax in 29 previously screened high-risk patients. All were examined within 24 hours, costing $272.07 per patient. Diagnoses were established quickly (median, same day; range, 0-15 days). Among 2259 at-risk postal workers, 144 (6.4%) self-identified new (< or =14 days) skin lesions and were examined in the field; 8 (5.6%) were referred to our system. Our system was not the only local dermatologic resource available during the outbreak. A system featuring initial nondermatologist examination with minimal laboratory evaluation, followed by rapid centralized referral of high-risk patients, functioned efficiently in this outbreak.

Adolescent↗

PubMed2006

Smallpox vaccines: Past, present, and future.

The global eradication of smallpox was a tremendous achievement made possible by the development of an effective vaccine. Routine vaccination of the general population is no longer recommended. However, stocks of variola virus, the causative agent of smallpox, still exist in 2 secure laboratories, and permanent disposal has been controversial. In addition, there is speculation that variola virus may exist outside of these 2 facilities, and there is a concern that the threat of smallpox will be used as a bioterrorist weapon. In 2002, this concern led to a vaccination campaign in US military and civilian healthcare workers and first responders. Although the historical live virus vaccine has proven efficacy, it also is associated with serious adverse events and rare fatal reactions, particularly in the setting of immunodeficiency and atopic eczema. In addition, this vaccine was historically produced using animal intermediaries in a process that was prone to contamination and not acceptable for current manufacturing standards. Development of alternative poxvirus vaccines is focused on replication-defective viruses, gene-based vectors, and subunit approaches to improve safety and immunogenicity. The conundrum is that in the absence of an intentional release of variola, efficacy evaluation of new candidate vaccines will be limited to animal model testing, which creates new challenges for the vaccine licensure process. Although motivated by the threat of bioterrorism, the hope is for new poxvirus vaccines to have their greatest utility against other pathogenic orthopoxviruses such as monkeypox and for the development of recombinant poxvirus-based vectors to treat and prevent other diseases.

Animals↗

PubMed2005

Algorithms for rapid outbreak detection: a research synthesis.

The threat of bioterrorism has stimulated interest in enhancing public health surveillance to detect disease outbreaks more rapidly than is currently possible. To advance research on improving the timeliness of outbreak detection, the Defense Advanced Research Project Agency sponsored the Bio-event Advanced Leading Indicator Recognition Technology (BioALIRT) project beginning in 2001. The purpose of this paper is to provide a synthesis of research on outbreak detection algorithms conducted by academic and industrial partners in the BioALIRT project. We first suggest a practical classification for outbreak detection algorithms that considers the types of information encountered in surveillance analysis. We then present a synthesis of our research according to this classification. The research conducted for this project has examined how to use spatial and other covariate information from disparate sources to improve the timeliness of outbreak detection. Our results suggest that use of spatial and other covariate information can improve outbreak detection performance. We also identified, however, methodological challenges that limited our ability to determine the benefit of using outbreak detection algorithms that operate on large volumes of data. Future research must address challenges such as forecasting expected values in high-dimensional data and generating spatial and multivariate test data sets.

Algorithms↗

PubMed2005

For debate: is disinfection of specimens, which may contain unknown or bio-terrorist organisms, essential before electron microscopic examination?

Severe and unexpected infections may be due to bioterrorism (BT), or the natural emergence of novel micro-organisms. Whatever the cause, there will be an urgent need to identify it for several reasons: defusing public anxiety, providing logical management and countermeasures, etc. For viruses and some bacteria, electron microscopy provides the fastest answers and identifying the cause quickly may eliminate BT. In the interests of safety, many would recommend that the specimen be disinfected at some stage before examination, but with unknown agents, however, reliable disinfection cannot be guaranteed and attempting to do so may also impair structure sufficiently to make recognition difficult or even impossible. As a basis for debate, this paper discusses the pros and cons of disinfecting such specimens.

Bioterrorism↗

PubMed2006

A susceptible-infected model of early detection of respiratory infection outbreaks on a background of influenza.

The threat of biological warfare and the emergence of new infectious agents spreading at a global scale have highlighted the need for major enhancements to the public health infrastructure. Early detection of epidemics of infectious diseases requires both real-time data and real-time interpretation of data. Despite moderate advancements in data acquisition, the state of the practice for real-time analysis of data remains inadequate. We present a nonlinear mathematical framework for modeling the transient dynamics of influenza, applied to historical data sets of patients with influenza-like illness. We estimate the vital time-varying epidemiological parameters of infections from historical data, representing normal epidemiological trends. We then introduce simulated outbreaks of different shapes and magnitudes into the historical data, and estimate the parameters representing the infection rates of anomalous deviations from normal trends. Finally, a dynamic threshold-based detection algorithm is devised to assess the timeliness and sensitivity of detecting the irregularities in the data, under a fixed low false-positive rate. We find that the detection algorithm can identify such designated abnormalities in the data with high sensitivity with specificity held at 97%, but more importantly, early during an outbreak. The proposed methodology can be applied to a broad range of influenza-like infectious diseases, whether naturally occurring or a result of bioterrorism, and thus can be an integral component of a real-time surveillance system.

Algorithms↗

PubMed2005

[Tularemia. The disease and its epidemiology in France].

OBJECTIVE: This study had for aim to determine whether the epidemiologic features of animal and human tularemia observed in France could explain its re-emergence since 1993. MATERIALS AND METHODS: A retrospective study was made of outbreaks in animals and human cases and contamination. RESULTS: Twenty to sixty animal outbreaks were recorded every year but the number of cases was under-estimated. This indicates an increase in animal cases. Human cases or contaminations were associated with areas where animal cases had been recorded, except for a few cases more difficult to explain. Several factors are implicated: the disease is not known, except by hunters. Tularemia is more often described in hares but the disease may be present in other species. The disease and its forms should be documented for clinicians and biologists for an adequate clinical and bacteriological diagnosis. COMMENTS: Several factors in the epidemiology of the animal diseases have changed since 1993. It is necessary to be increasingly vigilant, for animal cases and human contamination. It is necessary to strengthen medical and veterinary supervision especially since Francisella tularensis may be used for bioterrorism.

Animals↗

PubMed2005

[Melioidosis].

Melioidosis is an emerging zoonosis, due to Burkholderia pseudomallei, which is a highly invasive, resistant, and resilient soil bacteria, transmitted by cutaneous or airborne route, and is a potential weapon for bioterrorism. Although the agent has been identified all over the world, the human disease is endemic only in SE Asia and Northern Australia, and gained recent interest after the December 2004 tsunami. Human infection can be a very severe systemic disease (mortality 20 to 80%), with protean expression, but the lung is the most affected organ (50%). Pathophysiology remains unclear. Diabetes mellitus is a major risk factor, and is present in half the Asian patients with melioidosis. Recommended antibiotic regimens are expensive, and in severe disease should be prolonged to 20 weeks to reduce the risk of relapse. Prospects for prevention are limited, and no vaccine is available yet.

Australia↗

PubMed2004

Anthrax molecular epidemiology and forensics: using the appropriate marker for different evolutionary scales.

Precise identification of Bacillus anthracis isolates has aided forensic and epidemiological analyses of natural anthrax cases, bioterrorism acts and industrial scale accidents by state-sponsored bioweapons programs. Because there is little molecular variation among B. anthracis isolates, identifying and using rare variation is crucial for precise strain identification. We think that mutation is the primary diversifying force in a clonal, recently emerged pathogen, such as B. anthracis, since mutation rate is correlated with diversity on a per locus basis. While single nucleotide polymorphisms (SNPs) are rare, their detection is facilitated by whole genome discovery approaches. As highly stable phylogenetic markers, SNPs are useful for identifying long branches or key phylogenetic positions. Selection of single, diagnostic "Canonical SNPs" (canSNPs) for these phylogenetic positions allows for efficient and defining assays. We have taken a nested hierarchal strategy for subtyping B. anthracis, which is consistent with traditional diagnostics and applicable to a wide range of pathogens. Progressive hierarchical resolving assays using nucleic acids (PHRANA) uses a progression of diagnostic genomic loci that are initially highly stable but with low resolution and, ultimately, very unstable but with high resolution. This approach mitigates the need for data weighting and provides both a deeply rooted phylogenetic hypothesis and high resolution discrimination among closely related isolates.

Animals↗

PubMed2006

Rapid discrimination of Bacillus anthracis from other members of the B. cereus group by mass and sequence of "intact" small acid soluble proteins (SASPs) using mass spectrometry.

The intentional contamination of buildings, e.g. anthrax in the bioterrorism attacks of 2001, demonstrated that the population can be affected rapidly and lethally if the appropriate treatment is not provided at the right time. Molecular approaches, primarily involving PCR, have proved useful in characterizing "white powders" used in these attacks as well as isolated organisms. However there is a need for a simpler approach, which does not involve temperamental reagents (e.g. enzymes and primers) which could potentially be used by first responders. It is demonstrated here that small acid-soluble proteins (SASPs), located in the core region of Bacillus spores, are reliable biomarkers for identification. The general strategy used in this study was to measure the molecular weight (MW) of an intact SASP by electrospray ionization mass spectrometry (ESI MS) followed by generation of sequence-specific information by ESI MS/MS (tandem mass spectrometry). A prominent SASP of mass 6679 was present in all B. anthracis strains. For B. cereus and B. thuringiensis strains the SASP had a mass of 6712. This represents a two amino acid substitution (serine to alanine; phenylalanine to tyrosine). The only SASP present in the B. anthracis genome consistent with this sequence is encoded by the gene ssB. This protein has a predicted mass of 6810, presumably post-translational processing leads to loss of methionine (mass 131) generating a SASP of mass 6679. This study showed that intact SASPs can be used as a biomarker for identification of B. anthracis; the protocol is simple and rapid. Extrapolation of this approach might prove important for real-time biodetection.

Anthrax↗

PubMed2004

Crystal structure of 7,8-dihydropteroate synthase from Bacillus anthracis: mechanism and novel inhibitor design.

Dihydropterate synthase (DHPS) is the target for the sulfonamide class of antibiotics, but increasing resistance has encouraged the development of new therapeutic agents against this enzyme. One approach is to identify molecules that occupy the pterin binding pocket which is distinct from the pABA binding pocket that binds sulfonamides. Toward this goal, we present five crystal structures of DHPS from Bacillus anthracis, a well-documented bioterrorism agent. Three DHPS structures are already known, but our B. anthracis structures provide new insights into the enzyme mechanism. We show how an arginine side chain mimics the pterin ring in binding within the pterin binding pocket. The structures of two substrate analog complexes and the first structure of a DHPS-product complex offer new insights into the catalytic mechanism and the architecture of the pABA binding pocket. Finally, as an initial step in the development of pterin-based inhibitors, we present the structure of DHPS complexed with 5-nitro-6-methylamino-isocytosine.

Amino Acid Sequence↗

PubMed2004

The history of ricin, abrin and related toxins.

Ricin, abrin and related plant toxins have played interesting and important roles in the history of clinical medicine and biomedical research. The use of these proteins in medical treatment since ancient times is reviewed. Later the proteins played important roles in the early days of immunological research and some of the fundamental principles of immunology were discovered with toxic proteins of this group. During the last three decades the mechanism of action of the toxins was elucidated. This led to a major effort to target the toxins to malignant cells. Ricin has been used in bioterrorism. Recently, the toxins have played important roles as experimental models to elucidate the intracellular trafficking of endocytosed proteins.

Abrin↗

PubMed2006

Recombinant Salmonella vaccines for biodefence.

There is a requirement for vaccines to protect against pathogens that may be misused for bioterrorism or biowarfare purposes. In particular, biodefence vaccines are required that may be used for safe and easy immunisation of populations and that can rapidly induce mucosal immunity to provide protection at the lung surface against a range of airborne agents. To address this need, recombinant Salmonella vaccines are being developed. In this review, the technologies used, considerations needed, progress made, and future prospects for developing multivalent Salmonella-based vaccines for biodefence are discussed.

Biological Warfare↗