Microbial forensic analysis for deliberate, accidental and natural release of pathogens entail serious ramifications on human health. Surface sampling and transport for downstream verification of biological agents is posed with challenges including presence of background microbial consortium, delay in transport from remote location, and vulnerability of biomolecules to compromised cold chain. Glass surface pre-exposed to field conditions was spiked with S. typhi and sampled with swab. Storage of swab in VTM, PBS, alkaline lysis buffer, and under desiccated condition at RT and 10 °C for a period of 3 days indicated significantly higher DNA recovery for swabs stored under desiccated conditions. Recovery of bacterial protein from three transport media indicated pronounced stability at both temperatures. Tape-lift was envisaged as the method of surface sampling for providing stability to analytes during transport. Post-spiking (S. typhi) incubation of adhesive tapes for 3 and 7 days at RT indicated DNA recovery on both the time points and extracted DNA was amenable to PCR amplification of target specific gene. Methodology was also optimized for recovery of bacterial protein from adhesive tapes using S. typhi and B. anthracis Sterne spores. Protein stability was observed for both the agents after 3 and 7 days of storage at RT. The proteins extracted from the spiked adhesive tapes could be successfully identified using high resolution mass spectrometry (HRMS) with significant proteome coverage (84% on 3 d and 60% on 7 d). Application of adhesive tapes for sampling smooth surfaces for extraction of bacterial nucleic acid and protein suggest stability of analytes even without cold chain.

Bacillus anthracis, a Gram-positive, spore-forming bacterium, poses significant public health risks due to its zoonotic potential and historical use in bioterrorism. Pathogenicity of B. anthracis is primarily attributed to its genetic components, including the essential plasmids pXO1 and pXO2. The pXO1 plasmid encodes toxic factors, while pXO2 facilitates immune evasion. The zoonotic potential, as well as potential bio-threat, necessitates investigation into novel therapeutic targets. Through metabolic network analysis, docking, and notably subtractive genomics, researchers have effectively pinpointed pathogen-specific target proteins essential for drug development. Subtractive genomics, in particular, stands out for its cost-effectiveness and ability to identify non-host homologous targets. In this study, we used a core proteome immunoinformatic approach to identify potential therapeutic targets in B. anthracis. A total of 3987 proteins were found to be conserved in B. anthracis and among them, 388 were essential, non-human homologous proteins. These proteins were then subjected to localization and pathway analysis which resulted in 79 unique pathway proteins. Through PPI and druggability analysis, we had identified 17 strain-independent therapeutic target proteins. All the identified proteins are non-homologous to human proteins and, therefore, are potential targets for drug development. These potential target proteins are involved primarily in cell division, peptidoglycan biosynthesis, and putative flagellar assembly, which are essential for bacterial survival and pathogenicity. These identified target proteins could be the foundation for the development of therapeutics against human anthrax.

Food defense encompasses both the prevention of accidental contamination and the protection of food and water systems against deliberate adulteration. Within this framework, algal toxins deserve greater attention because they occupy a critical intersection of water security, seafood and drinking-water safety, environmental change, supply-chain continuity, and public health preparedness. The novelty of this review lies in reframing algal toxins from a primarily environmental, seafood safety, or clinical toxicology topic into an integrated food and water defense problem. This perspective brings together harmful algal bloom ecology, toxin toxicology, seafood safety, drinking-water protection, preparedness for intentional adulteration, climate change risk, surveillance, and emergency response. Algal toxins can enter the human supply chain through multiple marine and freshwater pathways, are often undetectable by taste, odor, or appearance, may resist routine preparation measures, and can generate high-consequence disruptions even when contamination begins locally. The analysis further shows that prevention depends largely on upstream measures-including environmental surveillance, harvest-area restrictions, source-water protection, testing, supply-chain controls, and rapid public communication-rather than on consumer behavior. Framed in the context of the U.S. Food and Drug Administration's Intentional Adulteration Rule under the Food Safety Modernization Act, this paper situates algal toxins within a preventive defense model that integrates monitoring, vulnerability reduction, and emergency preparedness. Although most harmful algal bloom events are naturally occurring, algal toxins warrant attention in food-defense planning because they can contaminate seafood and drinking-water systems, complicate detection and attribution, and expose vulnerabilities across interconnected food and water infrastructures.

Francisella tularensis, a virulent bioterrorism threat, subverts host immune responses within macrophages, its primary replication niche. To elucidate the temporal dynamics of the host response, we announce comprehensive time-course RNA-seq data sets capturing the global transcriptional profile of murine RAW 264.7 macrophages across early and late stages of F. tularensis infection.

Francisella tularensis is a gram-negative bacterium that causes tularemia, a fatal zoonotic disease. F. tularensis has been used in the bioweapon programs of several countries. Its potential use as a bioterrorism agent led the CDC to classify F. tularensis as a Tier 1 Select Agent. The cytosolic sensor absent in melanoma 2 (Aim2) detects double-stranded DNA in the cytosol of infected cells and subsequently assembles a multiprotein complex known as the inflammasome. Inflammasome activation drives the secretion of IL-1β and IL-18, key pro-inflammatory cytokines required for controlling F. tularensis infection. Prior studies have shown that F. tularensis actively suppresses Aim2 inflammasome activation; however, the underlying mechanism remains unknown. We hypothesized that F. tularensis suppresses Aim2-mediated responses by modulating the intracellular redox environment. We utilized an F. tularensis live vaccine strain (LVS) mutant lacking OxyR (ΔoxyR), a transcriptional regulator that controls the expression of major antioxidant enzymes. Our results show that macrophages infected with the ΔoxyR mutant exhibit significantly higher levels of Aim2-dependent caspase-1 and IL-1β than those infected with wild-type bacteria. The expression of interferon regulatory factor 1 and the guanylate-binding proteins GBP2 and GBP5, upstream signaling components of the Aim2 inflammasome, is markedly higher in ΔoxyR-infected macrophages than in controls. These changes were absent in ΔoxyR-infected NADPH oxidase-deficient macrophages, which are unable to generate reactive oxygen species. Collectively, these findings demonstrate that the macrophage redox environment plays a key role in activating the Aim2 inflammasome. This work advances understanding of how F. tularensis-encoded factors subvert host innate immune defenses.

This review examines the historical development of biological warfare from premodern contamination practices to twentieth-century state programs, post-9/11 bioterrorism, and the renewed prominence of biological risk in the COVID-19 era. Drawing on historical, policy, and public-health scholarship, it shows how disease came to be understood not only as a natural threat but also as a strategic instrument shaped by scientific knowledge, military organization, and administrative secrecy. The review traces the transition from empirical uses of contamination in siege warfare and colonial settings to laboratory-based weaponization in the 20th century, including German sabotage during the First World War, Imperial Japan's Unit 731, British anthrax trials on Gruinard Island, and the expansion of United States and Soviet biological warfare programs during the Cold War. It also evaluates the limits of international restraint through the 1925 Geneva Protocol and the 1972 Biological Weapons Convention, emphasizing the persistent problem of verification illustrated by the 1979 Sverdlovsk anthrax outbreak. The analysis then follows the shift from state arsenals to public fear through Aum Shinrikyo and the 2001 anthrax letters before situating epidemic and pandemic disease within the broader politics of biological vulnerability. COVID-19 and gain-of-function research are treated as cases of intensified scrutiny of biosafety, uncertainty, and public mistrust, rather than evidence for any single-origin hypothesis. The review concludes that biological warfare is best understood not solely as the deliberate use of pathogens but as a broader political and institutional phenomenon emerging where scientific capacity, strategic fear, concealment, and social vulnerability converge.

Tularemia, caused by Francisella tularensis, presents cycles of re-emergence in the Northern Hemisphere. In 2024-2025, increased incidence has been documented in Central Europe (Austria: 117 cases; Slovenia: 38 cases; France: 150 cases) and Spain (Castile and León: 206 cases) while the United States reported 220 cases in 2024. These data, combined with advances in molecular diagnostics and therapeutics, underscore the need for updated clinical knowledge. Tularemia is classified as a Category A bioterrorism agent by the CDC due to its extraordinary virulence; as few as 10 bacterial cells can establish infection, and untreated cases carry 30% mortality. Clinical manifestations range from localized forms (ulceroglandular, glandular) to systemic presentations with risk of progression without treatment. Diagnostic advances include PCR and culture-independent whole genome sequencing. The 2025 CDC guidelines establish fluoroquinolones and doxycycline as first-line therapeutic options. Despite over seven decades of research, no licensed human vaccine exists, and disease control depends on preventive measure. This review provides an updated synthesis of the microbiological, epidemiological, clinical, and therapeutic aspects of tularemia, with emphasis on early diagnosis given the organism's epidemic potential.

SUMMARYIn 1876, Robert Koch published "The Etiology of Anthrax Disease, based on the Developmental History of Bacillus anthracis." This seminal work laid the foundation for microbiology as a discipline. To commemorate the 150[th] anniversary of its publication, this article will review the history of B. anthracis, from its evolutionary origins to recent advancements in understanding its pathogenesis. For millions of years, B. anthracis existed in the environment as a unique pathogen whose lifestyle was heavily reliant on causing lethal disease in mammalian hosts. Beginning around 10,000 years ago, anthrax outbreaks have caused terrifying disease in humans and have wreaked havoc on livestock. The efforts to understand, treat, and prevent this disease have produced major breakthroughs in microbiology, medicine, and agriculture. In the modern era, despite still threatening livestock populations, "anthrax" has taken on a new meaning as a feared agent of bioterror. Continued scientific investigation into B. anthracis is not only critical in the prevention of the disease, but it promises to reveal new discoveries about the microbial world and its interactions with human hosts.

Smallpox, eradicated globally in 1980, remains a major biodefense concern due to retained Variola virus stocks and the absence of population immunity following cessation of routine vaccination. Monkeypox virus (MPXV), a related orthopoxvirus, has reemerged as a global public health threat, with major outbreaks in 2022 and renewed spread from 2024 onward.

Ricin, a potent protein toxin derived from Ricinus communis, is considered one of the most significant threats in cases of bioterrorism, homicide, suicide, and accidental acute poisoning. However, its clinical and pathological features in fatal cases have not been systematically summarized. This systematic review aimed to summarize the current evidence regarding the clinical manifestations, toxicological biomarkers, autopsy findings, and medico-legal aspects of fatal ricin poisoning. A comprehensive literature search was conducted in accordance with PRISMA guidelines. Original studies, case reports, and case series reporting detailed clinical, toxicological, or autopsy data following fatal or near-fatal human exposure to ricin were included in the review. The available evidence suggests a relatively consistent clinical course, characterized by rapid progression from mild, nonspecific gastrointestinal symptoms to hypovolemic shock, multi-organ failure, and refractory cardiovascular collapse. Ricinine was consistently detected in blood, urine, and postmortem tissues, supporting its role as a reliable biomarker of ricin exposure, although current evidence is insufficient to establish definitive correlations with poisoning severity or toxicokinetics. Reported autopsy findings commonly included hemolysis, hemorrhagic necrosis of the gastrointestinal tract, hepatic injury, renal tubular necrosis, and evidence supporting the presence of a capillary leak syndrome. Several specific medico-legal challenges emerged from the reviewed literature, including the need for the development of novel analytical methods with improved detection limits and quantification capabilities for ricinine, the interpretation of postmortem redistribution phenomena, the distinction between lethal and non-lethal exposures, and the differential diagnosis with other septic shock-like syndromes. Overall, the available evidence suggests a relatively consistent clinicopathological pattern associated with fatal ricin poisoning that can be corroborated through ricinine detection, while emphasizing the importance of early toxicological investigations, particularly when no history of exposure is available. These findings provide a concise and integrated body of evidence to support clinical management, inform forensic investigations, and enhance public health preparedness, while highlighting the need for standardized postmortem sampling protocols and further research into the toxicodynamics of ricin in humans.

The plant toxin ricin, derived from the castor bean plant Ricinus communis, is a major bioterrorism agent that requires analytical strategies to confirm both toxin identity and functionality. Ricin consists of an enzymatically active A chain (RTA) and a galactose-binding B chain (RTB) linked by an inter-chain disulfide bond. Conventional bottom-up mass spectrometry identifies ricin using proteotypic peptides from the individual chains but does not establish whether the toxin is present as an intact, active holotoxin. Here, we present an integrated workflow that enables structural and functional detection of ricin present at trace levels in complex matrices. The proteomic mass spectrometry assay combines targeted detection of multiple peptides from both chains with the challenging inter-chain disulfide-linked peptide, an unambiguous marker of intact ricin. Dramatic enhancement of sensitivity in complex matrices was achieved through optimized immunocapture with sheep anti-mouse magnetic beads, enabling ∼80% recovery of ricin and compatibility with nano-LC-MS/MS acquisition at concentrations below 1 ng/mL. The proteomic method matched the sensitivity of the parallel adenine-release assay, with limits of detection (LOD) down to 10 pg/mL for the A-, B-chain peptides and adenine. The most abundant linker peptide variants were detected at 500 pg/mL. Good repeatability was achieved, with average CV values below 20% across analyses and matrices. This approach enables simultaneous assessment of chain connectivity and biological activity, increasing confidence in identification relative to conventional approaches and providing unique insights into the toxin's functionality.

The ProVector® Biodegradable, visual Olfactory, and Chemical (BVOC™) vector control device was tested to attract and control mosquitoes, meet Department of Defense (DoD) thermal and camouflage criteria, and applicability for deployment by air and hand. The BVOC device is composed of non-toxic biodegradable composite materials that can be formed into various containers and sizes. The BVOC, with pesticides can be deployed from the air, by air gun, and by hand. It can be camouflaged to match the military operational environment and incorporate low thermal detectability, and incorporates biodegradable, visual, olfactory, and chemical components to attract and control mosquito populations and potentially other vectors. The BVOC offers several advantages over current technologies; 1) reduces the impact on the environment, 2) targets adult and larvae mosquitoes with less pesticide,as it is retained in a bait station and reduces impact on non-target organisms, 3) can be used for collection of mosquito species, and 4) reduces risk to military personnel by blending into the environment (visual and IR) and eliminates the need to refill or collect the device. The device is designed for the safety of military personnel and requires no specialized training. The BVOC can be used for control and surveillance of mosquitoes and other vectors from tree level to ground level, and water. The ProVector BVOC device can also be used to release the MEVLABS Multi-Bioagent Simulant and MEVLABS Infectious disease control reagents, and has both military and civilian applications.