Effect of Different Isolation Methods on Bioaerosol Distribution: An In Vitro Study.

Purpose: The purposes of this in vitro study were to evaluate the effect of three isolation methods to mitigate bioaerosols during stainless steel crown (SSC) preparations and assess the distribution of Streptococcus mutans by aerosolization in closed-room operatories. Methods: Melamine teeth coated in laboratory-grown S. mutans biofilm were prepared for SSCs using three different isolation methods. Agar plates were placed in five locations throughout the operatory and opened during each preparation as well as for 10 minutes immediately following to collect aerosolized S. mutans. Bacterial colonies were counted after incubating plates for 48 hours. Data were analyzed for differences between the isolation method and plate locations. Results: Bacterial colony counts for teeth prepared using high-volume evacuation suction (HVE) with dental dam (DD) isolation were statistically significantly higher than for those prepared using HVE with a DryShield®(DS) and HVE with no isolation at the assistant (A) (P<0.001), operator face shield (FS) (P<0.001), and patient (Pt) (P=0.002) locations. No significant differences were found among isolation methods for parent (Pa) or rear delivery (RD) locations. The location that produced the most bacterial colony counts using HVE with DD isolation was FS (P<0.001), followed by A (P=0.04), Pt (P<0.001), and RD and Pa (P<0.001). Counts produced from teeth prepared with DS isolation were significantly higher at the Pt location than the A (P<0.001), FS (P=0.002), RD (P<0.001), and Pa (P=0.008) locations. Conclusion: The use of dental dam with high-volume evacuation suction during stainless steel crown preparations increased bioaerosols near the procedure, while dental evacuation systems (DryShield®) may effectively limit their spread.

Reducing Transmission of Airborne Respiratory Pathogens: A New Beginning as the COVID-19 Emergency Ends.

BACKGROUND: In response to the COVID-19 pandemic, new evidence-based strategies have emerged for reducing transmission of respiratory infections through management of indoor air. OBJECTIVES: This paper reviews critical advances that could reduce the burden of disease from inhaled pathogens and describes challenges in their implementation. DISCUSSION: Proven strategies include assuring sufficient ventilation, air cleaning by filtration, and air disinfection by germicidal ultraviolet (UV) light. Layered intervention strategies are needed to maximize risk reduction. Case studies demonstrate how to implement these tools while also revealing barriers to implementation. Future needs include standards designed with infection resilience and equity in mind, buildings optimized for infection resilience among other drivers, new approaches and technologies to improve ventilation, scientific consensus on the amount of ventilation needed to achieve a desired level of risk, methods for evaluating new air-cleaning technologies, studies of their long-term health effects, workforce training on ventilation systems, easier access to federal funds, demonstration projects in schools, and communication with the public about the importance of indoor air quality and actions people can take to improve it. https://doi.org/10.1289/EHP13878.

Exploring toilet plume bioaerosol exposure dynamics in public toilets using a Design of Experiments approach.

Bioaerosols generated during toilet flushing can contribute to the spread of airborne pathogens and cross-contamination in indoor environments. This presents an increased risk of fomite-mediated or aerosol disease transmission. This study systematically investigated the factors contributing to increased bioaerosol exposure following toilet flushing and developed an empirical model for predicting the exposure-relevant bioaerosol concentration. Air in a toilet cubicle was sampled by impaction after seeding with Clostridium difficile spores. Design of Experiments (DoE) main effects screening and full factorial design approaches were then employed to investigate the significant factors that heighten the risk of exposure to bioaerosols post-flush. Our findings reveal that the inoculated bacterial concentration (C), time elapsed after flushing (t), lateral distance (d), and mechanical ventilation (v) are significant predictors of bioaerosol concentration, with p-values < 0.05. The interaction term, C × d showed a marked increase in bioaerosol concentration up to 232 CFU/m3 at the closest proximity and highest pathogen load. The interplay of C and t (C × t) demonstrated a time-dependent attenuation of bioaerosol viability, with concentrations peaking at 241 CFU/m3 immediately post-flush and notably diminishing over time. The lateral distance and time post-flush (d × t) interaction also revealed a gradual decrease in bioaerosol concentration, highlighting the effectiveness of spatial and temporal dilution in mitigating bioaerosol exposure risks. Furthermore, there is an immediate rise in relative humidity levels post-flush, impacting the air quality in the toilet environment. This study not only advances our understanding of exposure pathways in determining bioaerosol exposure, but also offers pivotal insights for designing targeted interventions to reduce bioaerosol exposure. Recommendations include designing public toilets with antimicrobial surfaces, optimizing ventilation, and initiating timely disinfection protocols to prioritise surfaces closest to the toilet bowl during peak exposure periods, thereby promoting healthier indoor environments and safeguarding public health in high-traffic toilet settings.

Microbial dynamics and climatic interactions in pig sheds: Insights into airborne microbes and particulate matter concentrations.

Emissions of airborne pollutants from livestock buildings affect indoor air quality, the health and well-being of farmers, animals and the environment. This study aimed to evaluate the microbial count within pig sheds and its relationship with meteorological variables (temperature, relative humidity and air velocity) and particulate matter (PM10 and PM2.5) and microbial diversity. Sampling was conducted both inside and outside of two pig sheds over three seasons (summer, rainy and winter), with regular monitoring at fortnightly intervals. Results showed that the bacterial and fungal counts ranged from 0.07 to 3.98 x 103 cfu/m3 inside the sheds and 0.01 to 1.82 x 103 cfu/m3 outside. Seasonal variations were observed, with higher concentrations of particulate matter detected during the winter season, followed by summer. Climatic variables such as temperature, air velocity and relative humidity demonstrated significant impacts on the abundance of Enterobacteriaceae and fungi, while air velocity specifically influenced the presence of mesophilic bacteria and staphylococci. Importantly, no significant disparities were found between microbial counts and particulate matter levels. Staphylococcaceae emerged as the predominant bacterial family, while Aspergillus and Cladosporium spp. were the dominant fungal species within the pig sheds. The average levels of airborne bacteria and fungi in pig sheds were found to be within the recommended range, which can be attributed to the loose housing design and lower animal population on the farms.

Fungal and bacterial species on biowaste workers' hands and inhalation zone, and potential airway deposition.

This study aims to investigate the microbiological working environment of biowaste workers, focusing on airborne fungal and bacterial species exposure, size distribution, and species on workers' hands. The research, conducted across six plants with 45 personal exposure assessments, revealed a total of 150 bacterial species and 47 fungal species on workers' hands, including 19 and 9 species classified in risk class 2 (RC2), respectively. Workers' exposure analysis identified 172 bacterial and 32 fungal species, with several in RC2. In work areas, 55 anaerobic bacterial species belonging to RC2 were found. Different species compositions were observed in various particle size fractions, with the highest species richness for anaerobic bacteria in the fraction potentially depositing in the secondary bronchi and for fungi in the pharynx fraction. The geometric mean aerodynamic diameter (DG) of RC2 anaerobic bacteria was 3.9 µm, <1.6 µm for Streptomyces, 3.4 µm for Aspergillus, and 2.0 µm for Penicillium. Overlapping species were identified on workers' hands, in their exposure, and in work areas, with Bacillus amyloliquefaciens, Leuconostoc mesenteroides, Bacillus cereus, Enterococcus casseliflavus, and Aspergillus niger consistently present. While the majority of RC2 bacterial species lacked documented associations with occupational health problems, certain bacteria and fungi, including Bacillus cereus, Escherichia coli, Enterobacter, Klebsiella pneumonia, Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, Lichtheimia corymbifera, Lichtheimia ramosa, and Paecilomyces variotii, have previously been linked to occupational health issues. In conclusion, biowaste workers were exposed to a wide range of microorganisms including RC2 species which would deposit in different parts of the airways.

The Development of a Specific Nanofiber Bioreceptor for Detection of Escherichia coli and Staphylococcus aureus from Air.

Polluted air and the presence of numerous airborne pathogens affect our daily lives. The sensitive and fast detection of pollutants and pathogens is crucial for environmental monitoring and effective medical diagnostics. Compared to conventional detection methods (PCR, ELISA, metabolic tests, etc.), biosensors bring a very attractive possibility to detect chemicals and organic particles with the mentioned reliability and sensitivity in real time. Moreover, by integrating nanomaterials into the biosensor structure, it is possible to increase the sensitivity and specificity of the device significantly. However, air quality monitoring could be more problematic even with such devices. The greatest challenge with conservative and sensing methods for detecting organic matter such as bacteria is the need to use liquid samples, which slows down the detection procedure and makes it more difficult. In this work, we present the development of a polyacrylonitrile nanofiber bioreceptor functionalized with antibodies against bacterial antigens for the specific interception of bacterial cells directly from the air. We tested the presented novel nanofiber bioreceptor using a unique air filtration system we had previously created. The prepared antibody-functionalized nanofiber membranes for air filtration and pathogen detection (with model organisms E. coli and S. aureus) show a statistically significant increase in bacterial interception compared to unmodified nanofibers. Creating such a bioreceptor could lead to the development of an inexpensive, fast, sensitive, and incredibly selective bionanosensor for detecting bacterial polluted air in commercial premises or medical facilities.

First detection of triazole-resistant aspergillus fumigatus harbouring the TR34/L98H Cyp51A mutation in Burkina Faso.

BACKGROUND: Triazole-resistant Aspergillus fumigatus (TRAF) isolates are a growing public health problem with worldwide distribution. Epidemiological data on TRAF is limited in Africa, particularly in West Africa. OBJECTIVES: This study aimed to screen for the environmental presence of TRAF isolates in the indoor air of two hospitals in Burkina Faso. MATERIALS AND METHODS: Air samples were collected in wards housing patients at risk for invasive aspergillosis, namely infectious diseases ward, internal medicine ward, nephrology ward, pulmonology ward, medical emergency ward and paediatric ward. Sabouraud Dextrose Agar supplemented with triazoles was used to screen the suspected TRAF isolates and EUCAST method to confirm the resistance of suspected isolates. Sequencing of cyp51A gene was used to identify the resistance mechanism of confirmed TRAF isolates. RESULTS: Of the 198 samples collected and analysed, 67 showed growth of A. fumigatus isolates. The prevalence of TRAF isolates was 3.23% (4/124). One TRAF isolate exhibited a pan-triazole resistance. Sequencing of cyp51A gene identified the TR34/L98H mutation for this pan-triazole resistant isolate. This study showed for the first time the circulation of the pan-azole resistant isolate harbouring the TR34/L98H mutation in Burkina Faso. CONCLUSIONS: These findings emphasise the need to map these TRAF isolates in all parts of Burkina Faso and to establish local and national continuous surveillance of environmental and clinical TRAF isolates in this country.

Fungal diversity differences in the indoor dust microbiome from built environments on earth and in space.

Human occupied built environments are no longer confined to Earth. In fact, there have been humans living and working in low-Earth orbit on the International Space Station (ISS) since November 2000. With NASA's Artemis missions and the age of commercial space stations set to begin, more human-occupied spacecraft than ever will be in Earth's orbit and beyond. On Earth and in the ISS, microbes, especially fungi, can be found in dust and grow when unexpected, elevated moisture conditions occur. However, we do not yet know how indoor microbiomes in Earth-based homes and in the ISS differ due to their unique set of environmental conditions. Here we show that bacterial and fungal communities are different in dust collected from vacuum bags on Earth and the ISS, with Earth-based homes being more diverse (465 fungal OTUs and 237 bacterial ASVs) compared to the ISS (102 fungal OTUs and 102 bacterial ASVs). When dust from these locations were exposed to varying equilibrium relative humidity conditions (ERH), there were also significant fungal community composition changes as ERH and time elevated increased (Bray Curtis: R2 = 0.35, P = 0.001). These findings can inform future spacecraft design to promote healthy indoor microbiomes that support crew health, spacecraft integrity, and planetary protection.

The abundance and pathogenicity of microbes in automobile air conditioning filters across the typical cities of China and Europe.

Bioaerosols are widely distributed in urban air and can be transmitted across the atmosphere, biosphere, and anthroposphere, resulting in infectious diseases. Automobile air conditioning (AAC) filters can trap airborne microbes. In this study, AAC filters were used to investigate the abundance and pathogenicity of airborne microorganisms in typical Chinese and European cities. Culturable bacteria and fungi concentrations were determined using microbial culturing. High-throughput sequencing was employed to analyze microbial community structures. The levels of culturable bioaerosols in Chinese and European cities exhibited disparities (Analysis of Variance, P < 0.01). The most dominant pathogenic bacteria and fungi were similar in Chinese (Mycobacterium: 18.2-18.9 %; Cladosporium: 23.0-30.2 %) and European cities (Mycobacterium: 15.4-37.7 %; Cladosporium: 18.1-29.3 %). Bartonella, Bordetella, Alternaria, and Aspergillus were also widely identified. BugBase analysis showed that microbiomes in China exhibited higher abundances of mobile genetic elements (MGEs) and biofilm formation capacity than those in Europe, indicating higher health risks. Through co-occurrence network analysis, heavy metals such as zinc were found to correlate with microorganism abundance; most bacteria were inversely associated, while fungi exhibited greater tolerance, indicating that heavy metals affect the growth and reproduction of bioaerosol microorganisms. This study elucidates the influence of social and environmental factors on shaping microbial community structures, offering practical insights for preventing and controlling regional bioaerosol pollution.

Global Spore Sampling Project: A global, standardized dataset of airborne fungal DNA.

Novel methods for sampling and characterizing biodiversity hold great promise for re-evaluating patterns of life across the planet. The sampling of airborne spores with a cyclone sampler, and the sequencing of their DNA, have been suggested as an efficient and well-calibrated tool for surveying fungal diversity across various environments. Here we present data originating from the Global Spore Sampling Project, comprising 2,768 samples collected during two years at 47 outdoor locations across the world. Each sample represents fungal DNA extracted from 24 m3 of air. We applied a conservative bioinformatics pipeline that filtered out sequences that did not show strong evidence of representing a fungal species. The pipeline yielded 27,954 species-level operational taxonomic units (OTUs). Each OTU is accompanied by a probabilistic taxonomic classification, validated through comparison with expert evaluations. To examine the potential of the data for ecological analyses, we partitioned the variation in species distributions into spatial and seasonal components, showing a strong effect of the annual mean temperature on community composition.

Association among polydisperse aerodynamic size of bioaerosols, biodiversity and urbanization in kindergartens.

The aerodynamic sizes of bioaerosols may significantly affect their behaviors, respiratory deposition and biodiversity. The respirable bacterial size, biodiversity, and human-associated bacteria (HAB) related bioaerosols were evaluated at three kindergartens in Taiwan. Kindergartens A, B, and C were in urban, semi-urban, and rural areas, respectively. A six-stage viable Andersen cascade impactor was used to collect bioaerosols and to determine their size distributions. The geometric mean diameter (GMD), geometric standard deviation (GSD), heat maps, and uniformity were used to evaluate the association of bacteria characteristics. A BD Phoenix-100 automated interpretation system was used to identify the airborne bacteria species. The results revealed that 1425 colonies of the sampled airborne bacteria contained 63 species in 29 genera, and overall, 63.0% were HABs. The most abundant phylum was Actinobacteria (56.6 ± 22.2%) and Firmicutes (31.6 ± 22.3%), and from the taxonomic analysis, both airborne Micrococcus and the Staphylococcus aureus are the dominant genus. All the bacteria aerodynamic particle size distributions were polydisperse distributions. The heat map and uniformity analysis had revealed most of the sampled bioaerosols distributed between 1.1-3.3 µm, and most of the polydisperse airborne Streptococcus spp. had a size in the respirable range, due to urbanization, they have potentially contributed to respiratory risk in the kindergartens. The Shannon diversity index (H) and inverse Simpson diversity index (D) of the bioaerosols in urban kindergarten were negatively correlated with GMD and GSD. The Pearson correlations revealed that the kindergarten in the rural area, with a higher temperature, a lower relative humidity, and a lower CO2 concentration than the others, tended to have the largest H and D values (P < 0.05). Multiple and stepwise regression revealed that bioaerosol aerodynamic size was statistically significantly correlated with H (P = 0.001) and D values (P = 0.002). This study sheds light on the characteristics of bioaerosols and their associations with microbiome.

Characteristics of bioaerosol emissions from a municipal wastewater treatment plant: Health risk assessment and microbial composition.

Bioaerosols released from municipal wastewater treatment plants (MWWTPs) contain pathogenic microorganisms, if dispersed into the atmosphere, which pose potential health risks to humans. In this study, the concentrations and size distribution of bioaerosol, factors on the bioaerosol emission, exposure risk, and microbial composition in different treatment units of a MWWTP were investigated. The results showed that bioaerosol was released to different degrees in each treatment unit, with the concentrations of bioaerosol varied widely, ranging from 978 to 3710 CFU/m3. FG and PST were primary bioaerosol emission sources in MWWTP. COD concentration, wind speed (WS) and relative humidity (RH) significantly influenced bioaerosol concentrations. The proportion of inhalable particles (< 4.7 µm) ranged from 51.35 % to 83.33 %, and bioaerosol emitted from WWTP caused a non-carcinogenic risk to children by the exposure risk assessment (HI > 1), which need to be paid more attention. Bacterial, fungal and actinomycete aerosols were detected in each treatment unit of MWWTP. Among these bioaerosols, bacterial aerosol was dominant. Importantly, several pathogenic bacteria including Sphingobium, Brevundimonas, Romboutsia, Arcobacter, Acinetobacter, and Mycobacterium were identified within the airborne bacteria population, most of which originated from wastewater or sludge, particularly in the ambient air of AeT. Pathogenic bacteria from MWWTP should be studied further to determine their long-term behavior and possible health risks.

Online monitoring system for qualitative and quantitative analysis of bioaerosols by combined ATP bioluminescence assay with loop-mediated isothermal amplification.

Rapid detection of airborne pathogens is crucial in preventing respiratory infections and allergies. However, technologies aiming to real-time analysis of microorganisms in air remain limited due to the sparse and complex nature of bioaerosols. Here, we introduced an online bioaerosol monitoring system (OBMS) comprised of integrated units including a rotatable stainless-steel sintered filter-based sampler, a lysis unit for extracting adenosine triphosphate (ATP), and a single photon detector-based fluorescence unit. Through optimization of the ATP bioluminescence method and establishment of standard curves between relative luminescence units (RLUs) and ATP as well as microbial concentration, we achieved simultaneous detection of bioaerosols' concentration and activity. Testing OBMS with four bacterial and two fungal aerosols at a sampling flow rate of 10 to 50 L/min revealed an outstanding collection efficiency of 95 % at 30 L/min. A single OBMS measurement takes only 8 min (sampling: 5 min; lysis and detection: 3 min) with detection limits of 3 Pcs/ms photons (2.9 × 103 and 292 CFU/m3 for Staphylococcus aureus and Candida albicans aerosol). In both laboratory and field tests, OBMS detected higher concentrations of bioaerosol compared to the traditional Andersen impactor and liquid biosampler. When combined OBMS with loop-mediated isothermal amplification (LAMP), the bioaerosol can be qualitative and quantitative analyzed within 40 min without the cumbersome procedures of sample pretreatment and DNA extraction. These results offer a high compressive and humidity resistance membrane filtration sampler and validate the potential of OBMS for online measurement of bioaerosol concentration and composition.

Bioaerosol emissions from wastewater treatment process at urban environment and potential health impacts.

The inlet of wastewater treatment plants (WWTPs) contains pathogenic microorganisms which during aeration and by mechanical mixing through wind typically aerosolized microbes into ambient air. Bioaerosol emission and its characterization (bacterial and fungal) was investigated considering low-flow and high-flow inlet of wastewater treatment plant. Generation of bioaerosols was found influenced by prevailing seasons while both during summer and winter, fungal concentration (winter: 1406 ± 517; summer: 1743 ± 271 CFU/m3) was higher compared to bacterial concentration (winter: 1077 ± 460; summer: 1415 ± 588 CFU/m3). Bioaerosols produced from WWTPs were predominately in the size range of 2.1-4.7 µm while fraction of fungal bioaerosols were also in ultra-fine range (0.65 µm). Bioaerosols reaching to the air from WWTPs varied seasonally and was calculated by aerosolization ratio. During summer, aerosolization of the bioaerosols was nearly 6 times higher than winter. To constitute potential health effects from the exposure to these bioaerosols, biological characterization, antibiotics resistance and the health survey of the nearby area were also performed. The biological characterization of the bioaerosols samples were done through metagenomic approach using 16s and ITS metagenomic sequencing. Presence of 167 genus of bacteria and 41 genus of fungi has been found. Out of this, bacillus (73%), curtobacterium (21%), pseudomonas, Exiguo bacterium, Acinetobacter bacillaceae, Enterobacteriaceae and Prevotella were the dominant genus (top 10) of bacteria. In case of fungi, xylariales (49%), Hypocreales (19%), Coperinopsis (9%), Alternaria (8%), Fusarium (6%), Biopolaris, Epicoccum, Pleosporaceae, Cladosporium and Nectriaceae were dominant. Antibiotics like, Azithromycin and cefixime were tested on the most dominant bacillus showed resistance on higher concentration of cefixime and lower concentration of azithromycin. Population-based health survey in WWTP nearby areas (50-150 m periphery) found several types of diseases/symptoms including respiratory problem, skin rash/irritation, change in smell and taste, eye irritation within the resident population and workers.

Diel variations of airborne microbes and antibiotic resistance genes in Response to urban PM2.5 chemical properties during the heating season.

Among the components of fine particulate matter (PM2.5), the contributions of airborne microorganisms and antibiotic resistance genes (ARGs) to health risks have been overlooked. Airborne microbial dynamics exhibit a unique diurnal cycle due to environmental influences. However, the specific roles of PM2.5 chemical properties resulting from fossil fuel combustion in driving circadian fluctuations in microbial populations and ARGs remain unclear. This study explored the interactions between toxic components and microbial communities during the heating period to understand the variations in ARGs. Bacterial and fungal communities showed a higher susceptibility to diel variations in PM2.5 compared to their chemical properties. Mantel tests revealed that chemical properties and microbial community interactions contribute differently to ARG variations, both directly and indirectly, during circadian fluctuations. Our findings highlight that, during the daytime, the enrichment of pathogenic microorganisms and ARGs increases the risk of PM2.5 toxicity. Conversely, during the nighttime, the utilization of water-soluble ions by the fungal community increased, leading to a significant increase in fungal biomass. Notably, Aspergillus exhibited a significant correlation with mobile genetic elements and ARGs, implying that this genus is a crucial driver of airborne ARGs. This study provides novel insights into the interplay between the chemical composition, microbial communities, and ARGs in PM, underscoring the urgent need for a comprehensive understanding of effective air pollution control strategies.

VLP-based model for the study of airborne viral pathogens.

The recent COVID-19 pandemic has underscored the danger of airborne viral pathogens. The lack of model systems to study airborne pathogens limits the understanding of airborne pathogen distribution as well as potential surveillance and mitigation strategies. In this work, we develop a novel model system to study airborne pathogens using virus-like particles (VLPs). Specifically, we demonstrate the ability to aerosolize VLP and detect and quantify aerosolized VLP RNA by reverse transcription-loop-mediated isothermal amplification in real-time fluorescent and colorimetric assays. Importantly, the VLP model presents many advantages for the study of airborne viral pathogens: (i) similarity in size and surface components; (ii) ease of generation and noninfectious nature enabling the study of biosafety level 3 and biosafety level 4 viruses; (iii) facile characterization of aerosolization parameters; (iv) ability to adapt the system to other viral envelope proteins, including those of newly discovered pathogens and mutant variants; and (v) the ability to introduce viral sequences to develop nucleic acid amplification assays. IMPORTANCE: The study and detection of airborne pathogens are hampered by the lack of appropriate model systems. In this work, we demonstrate that noninfectious virus-like particles (VLPs) represent attractive models to study airborne viral pathogens. Specifically, VLPs are readily prepared, are similar in size and composition to infectious viruses, and are amenable to highly sensitive nucleic acid amplification techniques.

Gel-confined fabrication of fully bio-based filtration membrane for green capture and rapid detection of airborne microbes.

Air filtration has become a desirable route for collecting airborne microbes. However, the potential biotoxicity and sterilization of current air filtration membranes often lead to undesired inactivation of captured microbes, which greatly limits microbial non-traumatic transfer and recovery. Herein, we report a gel-confined phase separation strategy to rationally fabricate a fully bio-based filtration membrane (SGFM) using soluble soybean polysaccharide and gelatin. The versatile SGFM features fascinating honeycomb micro-nano architecture and hierarchical interconnected porous structures for microbial capture, and achieves a lower pressure drop, higher interception efficiency (99.3%), and superior microbial survivability than commercial gelatin filtration membranes. Particularly, the water-dissolvable SGFM can greatly simplify the elution and extraction process after bioaerosol sampling, thereby bringing about maximum sample transfer and vigorous recovery of collected microbes. Meanwhile, green capture coupled with ATP bioluminescence endows the SGFM with rapid and quantitative detection capability for airborne microbes. This work may pave the way for designing green protocols for the detection of bioaerosols.

Sampling efficiency and nucleic acid stability during long-term sampling with different bioaerosol samplers.

Aerosol microbiome studies have received increased attention as technological advancements have made it possible to dive deeper into the microbial diversity. To enhance biomass collection for metagenomic sequencing, long-term sampling is a common strategy. While the impact of prolonged sampling times on microorganisms' culturability and viability is well-established, its effect on nucleic acid stability remains less understood but is essential to ensure representative sample collection. This study evaluated four air samplers (SKC BioSampler, SASS3100, Coriolis µ, BioSpot-VIVAS 300-P) against a reference sampler (isopore membrane filters) to identify nucleic acid stability during long-term sampling. Physical sampling efficiencies determined with a fluorescent tracer for three particle sizes (0.8, 1, and 3 µm), revealed high efficiencies (> 80% relative to reference) for BioSampler, SASS3100, and BioSpot-VIVAS for all particle sizes, and for Coriolis with 3 µm particles. Coriolis exhibited lower efficiency for 0.8 µm (7%) and 1 µm (50%) particles. During 2-h sampling with MS2 and Pantoea agglomerans, liquid-based collection with Coriolis and BioSampler showed a decrease in nucleic acid yields for all test conditions. BioSpot-VIVAS displayed reduced sampling efficiency for P. agglomerans compared to MS2 and the other air samplers, while filter-based collection with SASS3100 and isopore membrane filters, showed indications of DNA degradation for 1 µm particles of P. agglomerans after long-term sampling. These findings show that long-term air sampling affects nucleic acid stability in both liquid- and filter-based collection methods. These results highlight bias produced by bioaerosol collection and should be considered when selecting an air sampler and interpreting aerosol microbiome data.

A super sandstorm altered the abundance and composition of airborne bacteria in Beijing.

Sandstorm, which injects generous newly emerging microbes into the atmosphere covering cities, adversely affects the air quality in built environments. However, few studies have examined the change of airborne bacteria during severe sandstorm events. In this work, we analyzed the airborne bacteria during one of the strongest sandstorms in East Asia on March 15th, 2021, which affected large areas of China and Mongolia. The characteristics of the sandstorm were compared with those of the subsequent clean and haze days. The composition of the bacterial community of air samples was investigated using quantitative polymerase chain reaction (qPCR) and high-throughput sequencing technology. During the sandstorm, the particulate matter (PM) concentration and bacterial richness were extremely high (PM2.5: 207 µg/m3; PM10: 1630 µg/m3; 5700 amplicon sequence variants/m3). In addition, the sandstorm brought 10 pathogenic bacterial genera to the atmosphere, posing a grave hazard to human health. As the sandstorm subsided, small bioaerosols (0.65-1.1 µm) with a similar bacterial community remained suspended in the atmosphere, bringing possible long-lasting health risks.