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.

[This corrects the article DOI: 10.3389/fpubh.2026.1792476.].

Inhaling Bacillus anthracis spore-laden dust or aerosols can cause inhalational anthrax; such direct-inhalation infection is uncommon and usually affects people who handle diseased animals or animal products.

Anthrax is a zoonotic disease caused by the bacterium Bacillus anthracis (B. anthracis). Inhalation anthrax, caused by inhalation of B. anthracis spores, is the rarest and most fatal form of anthrax. Historically, naturally acquired inhalation anthrax cases have mainly been associated with occupational exposure to infected animals or animal products, such as in wool mills or tanneries.

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.

A combined molecular strategy was developed to accurately differentiate wild-type and vaccine strains of Bacillus anthracis. This approach integrates a multiplex TaqMan qPCR assay targeting the BclA, sap, and cap genes with conventional PCR analysis of BclA fragment length polymorphism. The qPCR assay demonstrated high specificity, with no cross-reactivity against other common pathogens, and exhibited a consistent limit of detection of 2.11 × 10[1] copies/μL (Cq < 35) for all three targets. Excellent repeatability was confirmed, with intra- and inter-assay coefficients of variation below 1.96% and 3.06%, respectively. Validation using 19 wild-type strains revealed two distinct qPCR profiles, and all wild-type strains were conclusively differentiated from the capsule-producing Chinese vaccine strain II and the acapsular Sterne strain by their characteristic BclA amplicon sizes. Through simulation PCR validation of common global canSNP genotyping (groups A, B, and C), it is possible to distinguish between wild-type and vaccine strains in China. This integrated "multiplex qPCR-BclA length analysis" protocol provides a reliable, sensitive, and specific tool for the precise detection and differentiation of B. anthracis strains in China.IMPORTANCEAnthrax, which is caused by the bacterium B. anthracis, remains a serious zoonotic disease. Its effective surveillance relies on precise strain identification. Currently, it is challenging to rapidly and accurately distinguish wild-type strains from vaccine strains, which hinders outbreak investigation and response. This study develops a novel molecular strategy combining multiple genetic targets to solve this critical diagnostic problem. The new method provides a reliable tool for strain differentiation, directly supporting accurate source tracing and informed decision-making during anthrax outbreaks. By improving diagnostic precision, this work contributes to strengthening public and veterinary health defenses against this persistent threat.

In the original publication [...].

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.

Environmental persistence of Bacillus anthracis spores sustains anthrax transmission, necessitating rapid and field-deployable detection tools. This study aimed to develop and quantitatively evaluate a LAMP-assisted CRISPR-Cas12a assay for sensitive and specific detection of B. anthracis spores in soil and meat meal matrices. Two B. anthracis-specific target genes plasmid-encoded lef and chromosomal SNP locus (CR5)- were selected. Target-specific LAMP primers and CRISPR crRNAs were designed using Primer Explorer, CRISPOR, and RNAfold platforms. The LAMP-CRISPR/Cas12a assay was optimised for reaction conditions and evaluated for analytical sensitivity and specificity using UV-inactivated spores and closely related Bacillus spp. Environmental applicability was assessed via spiking experiments in sterile soil and meat meal using the GABRI recovery method. Field validation was performed on 100 samples from anthrax-endemic regions of India, with performance compared against the WOAH-recommended lef gene-based real-time PCR. LAMP-CRISPR-Cas12a assay achieved detection limits of 10 spores/ml (lef) and 10[2] spores/ml (CR5), with no cross-reactivity against related species. In spiked matrices, spore recovery ranged from 50 to 75%, and detection sensitivity remained consistent. Field evaluation demonstrated a sensitivity of 91.70% and specificity of 100%, with near-perfect agreement (κ = 0.95) relative to real-time PCR. The assay delivered results within ∼70 min, including amplification and detection. The LAMP-assisted CRISPR-Cas12a platform provides a rapid, sensitive, and cost-effective approach for environmental detection of B. anthracis spores. The minimal equipment requirements and high diagnostic accuracy support its applicability for field-level surveillance and biosafety monitoring in resource-limited settings.

Anthrax lethal toxin is a key virulence factor of Bacillus anthracis. However, non-canonical epitopes of protective antigen (PA) and alternative neutralization mechanisms beyond receptor blockade or prevention of proteolytic cleavage remain underexplored. Here, we characterize a fully human monoclonal antibody, 22F1, with ultrapotent toxin neutralization in vitro (IC50 = 0.027 nM) and in vivo (molar ratio 1:7.5-15). Unlike known PA antibodies, 22F1 selectively binds the post-cleavage form PA63, not full-length PA83, due to shielding of the epitope by PA20. Cryo-EM reveals a novel conformational epitope spanning domains D1', D2, and D3. 22F1 does not block receptor engagement, PA proteolytic activation, or lethal factor binding. Instead, its epitope lies near the inter-protomer interface; binding of a single 22F1 Fab to any PA63 protomer exerts a dual steric hindrance effect, simultaneously preventing additional Fab binding to adjacent protomers and blocking PA63 oligomerization. This "single-hit" mechanism enables one antibody molecule to efficiently block prepore formation, achieving high potency. Our findings advance anthrax antitoxin research and provide a paradigm for targeting other proteins with similar activation-oligomerization-pore formation mechanisms.