Les Baillie

During construction of the Baku–Tbilisi–Kars railroad between Kars City in Türkiye and Tbilisi in Georgia, blasting operations in an anthrax-endemic region disrupted a burial pit containing carcasses of cattle that had died of anthrax. Railroad workers expressed concerns that release of this material could result in them developing anthrax infection. We therefore undertook a seroprevalence study six months later to seek evidence of exposure to Bacillus anthracis spores. We used an optimised Enzyme-Linked Immunosorbent Assay (ELISA) to screen serum for antigen-specific IgG antibodies to the anthrax toxin subunits Protective Antigen (PA) and Lethal Factor (LF). Stepwise linear regressions and t-tests were performed to compare results from railroad workers (n = 64) with a group of long-term Kars City residents (urban dwellers, n = 16), who had no history of possible contact with anthrax antigens. Anti-PA IgG concentrations were higher (p = 0.038) in railroad workers than in urban dwellers, but anti-LF IgG concentrations did not differ (p = 0.932) between the two groups. The anti-PA response is known to be dominant, and the difference was small. The lack of LF response did not preclude an antibody response to B. anthracis. Following the blasting operations, no cases of anthrax infection occurred in either railroad workers or villagers living nearby, suggesting that the spore exposure (evidenced by higher antibody titres) was at levels insufficient to initiate clinical infection. The elevated PA-specific antibody responses in railroad workers compared with urban dwellers might be consistent with the former having had previous subclinical exposure to B. anthracis. In anthrax-endemic regions, therefore, construction activities that involve blasting or large-scale excavation may pose risks of occupational exposure to Bacillus anthracis spores.

He workshop concluded that environmental conditions, spore characteristics, and surface types all influence how Bacillus anthracis behaves after decontamination. Factors such as UV exposure, humidity, pH, weathering, and soil type affect spore survival and adhesion, while particle size and attachment to dust or fibres shape inhalation risk. Porous materials, carpets, and busy environments increase the chance of resuspension. Although engineering controls can reduce the movement of residual spores, major knowledge gaps remain, especially regarding the infectious dose, post‑decontamination aerosol formation, and how exposure varies with height and population vulnerability. Effective recovery requires targeted, multi‑step decontamination using non‑persistent chemicals, with potential support from biological tools such as germinants or bacteriophages. Agricultural areas may need long‑term land‑use planning where complete removal is unrealistic. Access control, proportionate security, and reliable monitoring, particularly methods that distinguish viable spores from DNA, are essential. Clear, consistent communication is critical: The public must understand that zero spores cannot be guaranteed, and acceptable residual risk depends on context, land use, and operational practicality. Policies must balance scientific uncertainty, resource limits, population needs, and coordinated messaging while managing misinformation risks. Overall, the workshop urged a shift from unrealistic zero‑spore expectations toward flexible, evidence‑based approaches that balance science, practicality, and public expectations.

The aim of our research is to develop the environmentally friendly and reusable antimicrobial packaging based on the beeswax with antibacterial agents: propolis and sage extracts and evaluation of its storage abilities and antibacterial properties. During the experiment were used cotton fabrics, beeswax, propolis, sage leaves, emulsifiers and antioxidants were used as materials for the development of the packaging. Samples of beeswax wraps were produced using three technologies. The obtained samples were tested for food storage ability (for bread, cheese and sausages) and antimicrobial activity against S. aureus, E. coli, P. auroginosa and C. albicans. Obtained wraps samples have been tested on a basic set of products available in every household: bakery products, hard cheeses and ready-made sausages. Beeswax wrapping prepared by all methods was found to be the best for cheese storage. Results of the experiment on the determination of the antimicrobial activity of beeswax wraps and antimicrobial agents against S. aureus, E. coli, P. аeruginosa, C. albicans have shown that sage extract has high activity against S. aureus and moderate activity against E. coly and P. аeruginosa. Propolis oily extract possess moderate activity against tested microorganisms and both extracts had low antifungal activity. Determination of the antimicrobial activity of beeswax wraps and antimicrobial agents against S. aureus, E. coli, P. аeruginosa, C. albicans have shown moderate and low activity against microorganisms and no antifungal activity. Summing up we could conclude that beeswax wraps can act as a promising packaging for a food products.

Patient loss to follow-up caused by centralised and expensive diagnostics that are reliant on sputum is a major obstacle in the fight to end tuberculosis. An affordable, non-sputum biomarker-based, point-of-care deployable test is needed to address this. Serum antibodies binding the mycobacterial cell wall lipids, mycolic acids, have shown promise as biomarkers for active tuberculosis. However, anti-lipid antibodies are of low affinity, making them difficult to detect in a lateral flow immunoassay—a technology widely deployed at the point-of-care. Previously, recombinant monoclonal anti-mycolate antibodies were developed and applied to characterise the antigenicity of mycolic acid. We now demonstrate that these anti-mycolate antibodies specifically detect hexane extracts of mycobacteria. Secondary antibody-mediated detection was applied to detect the displacement of the monoclonal mycolate antibodies by the anti-mycolic acid antibodies present in tuberculosis-positive guinea pig and human serum samples. These data establish proof-of-concept for a novel lateral flow immunoassay for tuberculosis provisionally named MALIA—mycolate antibody lateral flow immunoassay.

Environmental contamination with Bacillus anthracis spores poses clear threats to livestock that play key roles in the economies of pastoral communities. Regular monitoring of contaminated sites is particularly important in anthrax-endemic parts of the world, such as Kars province in eastern Türkiye, where the Veterinary Microbiology Department of Kafkas University has conducted an anthrax surveillance programme for over 30 years. We reviewed the microbiological results of 232 soil samples collected during 2009–2023, from sites known to be contaminated with B. anthracis spores following burial or butchering of infected animal carcasses. Twenty-five contaminated sites in 16 villages were studied. Samples were taken from a total of 61 different positions within these sites and viable spores were detected in 136 (58.6%) of the samples examined. Of the 96 samples from which spores were not recovered, subsequent samples from the same positions proved positive on 21 occasions. Using a standardised sampling plan, it was discovered that samples taken 1–2 m on a downward slope from the centre-point of contamination had higher (p < 0.001) spore concentrations than those taken from other positions. Although spore concentrations at some sampling positions varied over time, the overall values remained stable. This finding contrasts with observations in other parts of the world where spore concentrations tend to decline with time and may reflect regional differences in soil composition that permit more prolonged spore persistence. Concentrations of >100 spores/g soil were found in 10 (66.7%) of the 15 samples taken 10–13 years following a contamination event. These results demonstrate the longevity of viable anthrax spores in the soil of agricultural environments following decomposition of infected animal carcasses, and therefore the need for prolonged bacteriological monitoring of contaminated sites. Furthermore, they underline the importance of appropriate decontamination, as burial on its own does not eliminate all spores.

The rapid detection of the spore form of Clostridioides difficile has remained a challenge for clinicians. To address this, we have developed a novel, precise, microwave-enhanced approach for near-spontaneous release of DNA from C. difficile spores via a bespoke microwave lysis platform. C. difficile spores were microwave-irradiated for 5 s in a pulsed microwave electric field at 2.45 GHz to lyse the spore and bacteria in each sample, which was then added to a screen-printed electrode and electrochemical DNA biosensor assay system to identify presence of the pathogen’s two toxin genes. The microwave lysis method released both single-stranded and double-stranded genome DNA from the bacterium at quantifiable concentrations between 0.02 μg/mL to 250 μg/mL allowing for subsequent downstream detection in the biosensor. The electrochemical bench-top system comprises of oligonucleotide probes specific to conserved regions within tcdA and tcdB toxin genes of C. difficile and was able to detect 800 spores of C. difficile within 300 µL of unprocessed human stool samples in under 10 min. These results demonstrate the feasibility of using a solid-state power generated, pulsed microwave electric field to lyse and release DNA from human stool infected with C. difficile spores. This rapid microwave lysis method enhanced the rapidity of subsequent electrochemical detection in the development of a rapid point-of-care biosensor platform for C. difficile.

This article was originally published online on 25 January 2024 with images and captions incorrectly labeled in Figs. 15, 16, and 17. Figure 15 has been changed

This paper describes the design and optimization of a 10 ml cartridge for patient sample processing using a 3.5 GHz (empty resonant frequency) TM010 cylindrical microwave cavity. The cartridge has been designed to augment a novel approach for the rapid diagnosis of M. tuberculosis (the causative agent of Ttuberculosis), which uses the direct application of microwaves to a bacteria-containing sample to release pathogen-specific DNA. The target bacterial DNA is then captured and recovered using magnetic nanoparticles coated with pathogen-specific DNA probes. Excitation parameters were optimized using three surrogates for M. tuberculosis, namely, M. smegmatis, M. abscessus, and M. bovis suspended in water and simulated sputum. The paper also explores the mechanism of microwave-mediated DNA release from bacteria using scanning electron microscopy. Examination of bacteria exposed to microwaves at power levels known to mediate the release of DNA reveals no obvious signs of permanent cell disruption, suggesting that a more subtle interaction is taking place. Finally, the presence of microwave-liberated M. bovis DNA was able to be detected at a level of sensitivity comparable to that achieved using microscopy.

Environmental contamination with Bacillus anthracis spores poses uncertain threats to human health. We undertook a study to determine whether inhabitants of the anthrax-endemic region of Kars in eastern Türkiye could develop immune responses to anthrax toxins without recognised clinical infection. We measured anti-PA and anti-LF IgG antibody concentrations by ELISA in serum from 279 volunteers, 105 of whom had previously diagnosed anthrax infection (100 cutaneous, 5 gastrointestinal). Of the 174 without history of infection, 72 had prior contact with anthrax-contaminated material. Individuals were classified according to demographic parameters, daily working environment, and residence type. All villages in this study had recorded previous animal or human anthrax cases. Stepwise regression analyses showed that prior clinical infection correlated strongly with concentrations at the upper end of the ranges observed for both antibodies. For anti-PA, being a butcher and duration of continuous exposure risk correlated with high concentrations, while being a veterinarian or shepherd, time since infection, and town residence correlated with low concentrations. For anti-LF, village residence correlated with high concentrations, while infection limited to fingers or thumbs correlated with low concentrations. Linear discriminant analysis identified antibody concentration profiles associated with known prior infection. Profiles least typical of prior infection were observed in urban dwellers with known previous infection and in veterinarians without history of infection. Four individuals without history of infection (two butchers, two rural dwellers) had profiles suggesting unrecognised prior infection. Healthy humans therefore appear able to tolerate low-level exposure to environmental B. anthracis spores without ill effect, but it remains to be determined whether this exposure is protective. These findings have implications for authorities tasked with reducing the risk posed to human health by spore-contaminated materials and environments.