Exposure to resistant fungi across working environments and time.

Antifungal resistance has emerged as a significant health concern with increasing reports of resistant variants in previously susceptible species. At present, little is known about occupational exposure to antifungal-resistant fungi. This study aimed to investigate Danish workers' occupational exposure to airborne fungi resistant to first-line treatment drugs. A retrospective study was performed on a unique collection of personal exposure samples gathered over a twenty-year period from Danish working environments, in sectors including agriculture, animal handling, waste management, and healthcare. A total of 669 samples were cultivated at 37 °C and fungal colonies were identified using MALDI-TOF MS. Subsequently, identification was confirmed by amplicon sequencing the genes of calmodulin and beta-tubulin to unveil potential cryptic species. Infectious fungi (495 isolates from 23 species) were tested for resistance against Itraconazole, Voriconazole, Posaconazole, and Amphotericin B. Working environments were highly variable in the overall fungal exposure, and showed vastly different species compositions. Resistance was found in 30 isolates of the species Aspergillus fumigatus (4 of 251 isolates), A. nidulans (2 of 13), A. niger complex (19 of 131), A. versicolor (3 of 18), and A. lentulus (2 of 2). Sequence analysis revealed several cryptic species within the A. niger complex including A. tubingensis, A. luchuensis, and A. phoenicis. Among the resistant A. fumigatus isolates, two contained the well-described TR34/L98H mutation in the cyp51A gene and promoter region, while the remainder harbored silent mutations. The results indicate that the working environment significantly contributes to exposure to resistant fungi, with particularly biofuel plant workers experiencing high exposure. Differences in the prevalence of resistance across working environments may be linked to the underlying species composition.

Work clothes as a vector for microorganisms: Accumulation, transport, and resuspension of microorganisms as demonstrated for waste collection workers.

Work clothes may act as a vector for the transport of microorganisms leading to second-hand exposure; however, this has not been studied in work environments. We investigated whether microorganisms accumulate on workers' clothes in environments with elevated microbial exposures, and whether they are transported with the clothes and subsequently resuspended to the air. To study this, we selected waste collection workers and potential transport of bacteria and fungi to waste truck cabs via clothes, and compared the microbial communities within truck cabs, in waste collection workers' personal exposure, and on clean T-shirts worn by the workers. Microbial communities were also investigated for the presence of potentially harmful microorganisms. Results showed that microorganisms accumulated in large quantities (GM = 3.69 × 105 CFU/m2/h for bacteria, GM = 8.29 × 104 CFU/m2/h for fungi) on workers' clothes. The concentrations and species composition of airborne fungi in the truck cabs correlated significantly with the accumulation and composition of fungi on clothes and correlated to concentrations (a trend) and species composition of their personal exposures. The same patterns were not found for bacteria, indicating that work clothes to a lesser degree act as a vector for bacteria under waste collection workers' working conditions compared to fungi. Several pathogenic or allergenic microorganisms were present, e.g.: Klebsiella oxytoca, K. pneumoniae, Proteus mirabilis, Providencia rettgeri, Pseudomonas aeruginosa, and Aspergillus fumigatus, A. glaucus, A. nidulans, A. niger, and various Penicillium species. The potential 'take-home' exposure to these microorganisms are of most concern for immunocompromised or atopic individuals or people with open wounds or cuts. In conclusion, the large accumulation of microorganisms on workers' clothes combined with the overlap between fungal species for the different sample types, and the presence of pathogenic and allergenic microorganisms forms the basis for encouragement of good clothing hygiene during and post working hours.

Hyperbaric oxygen treatment increases killing of aggregating Pseudomonas aeruginosa isolates from cystic fibrosis patients.

BACKGROUND: Pseudomonas aeruginosa is a major pathogen of the chronic lung infections in cystic fibrosis (CF) patients. These persistent bacterial infections are characterized by bacterial aggregates with biofilm-like properties and are treated with nebulized or intravenous tobramycin in combination with other antibiotics. However, the chronic infections are close to impossible to eradicate due to reasons that are far from fully understood. Recent work has shown that re­oxygenation of hypoxic aggregates by hyperbaric oxygen (O2) treatment (HBOT: 100% O2 at 2.8 bar) will increase killing of aggregating bacteria by antibiotics. This is relevant for treatment of infected CF patients where bacterial aggregates are found in the endobronchial secretions that are depleted of O2 by the metabolism of polymorphonuclear leukocytes (PMNs). The main objective of this study was to investigate the effect of HBOT as an adjuvant to tobramycin treatment of aggregates formed by P. aeruginosa isolates from CF patients. METHODS: The effect was tested using a model with bacterial aggregates embedded in agarose. O2 profiling was used to confirm re­oxygenation of aggregates. RESULTS: We found that HBOT was able to significantly enhance the effect of tobramycin against aggregates of all the P. aeruginosa isolates in vitro. The effect was attributed to increased O2 levels leading to increased growth and thus increased uptake of and killing by tobramycin. CONCLUSIONS: Re­oxygenation may in the future be a clinical possibility as adjuvant to enhance killing by antibiotics in cystic fibrosis lung infections.