Hunting for a viral proxy in bioaerosols of swine buildings using molecular detection and metagenomics.

There are limited biosecurity measures directed at preventing airborne transmission of viruses in swine. The effectiveness of dust mitigation strategies such as oil sprinkling, to decrease risk of airborne virus transmission are unknown. Metagenomics and qPCR for common fecal viruses were used to hunt for a ubiquitous virus to serve as a proxy when evaluating the efficiency of mitigation strategies against airborne viral infectious agents. Air particles were collected from swine buildings using high-volume air samplers. Extracted DNA and RNA were used to perform specific RT-qPCR and qPCR and analyzed by high-throughput sequencing. Porcine astroviruses group 2 were common (from 102 to 105 genomic copies per cubic meter of air or gc/m3, 93% positivity) while no norovirus genogroup II was recovered from air samples. Porcine torque teno sus virus were detected by qPCR in low concentrations (from 101 to 102 gc/m3, 47% positivity). Among the identified viral families by metagenomics analysis, Herelleviridae, Microviridae, Myoviridae, Podoviridae, and Siphoviridae were dominant. The phage vB_AviM_AVP of Aerococcus was present in all air samples and a newly designed qPCR revealed between 101 and 105 gc/m3 among the samples taken for the present study (97% positivity) and banked samples from 5- and 15-year old studies (89% positivity). According to the present study, both the porcine astrovirus group 2 and the phage vB_AviM_AVP of Aerococcus could be proxy for airborne viruses of swine buildings.

Air monitoring by nanopore sequencing.

While the air microbiome and its diversity are essential for human health and ecosystem resilience, comprehensive air microbial diversity monitoring has remained rare, so that little is known about the air microbiome's composition, distribution, or functionality. Here we show that nanopore sequencing-based metagenomics can robustly assess the air microbiome in combination with active air sampling through liquid impingement and tailored computational analysis. We provide fast and portable laboratory and computational approaches for air microbiome profiling, which we leverage to robustly assess the taxonomic composition of the core air microbiome of a controlled greenhouse environment and of a natural outdoor environment. We show that long-read sequencing can resolve species-level annotations and specific ecosystem functions through de novo metagenomic assemblies despite the low amount of fragmented DNA used as an input for nanopore sequencing. We then apply our pipeline to assess the diversity and variability of an urban air microbiome, using Barcelona, Spain, as an example; this randomized experiment gives first insights into the presence of highly stable location-specific air microbiomes within the city's boundaries, and showcases the robust microbial assessments that can be achieved through automatable, fast, and portable nanopore sequencing technology.

Uncovering wastewater treatment plants as possible sources of legionellosis clusters through spatial statistics approach and environmental analysis.

Wastewater treatment plants (WWTPs) are suspected reservoirs of Legionella pneumophila (Lp). The required aeration and mixing steps lead to the emission and dispersion of bioaerosols potentially harboring Lp. The aim of the project is to evaluate municipal WWTPs as a possible source of legionellosis through the statistical analysis of case clusters. A space-time scanning statistical method was implemented in SaTScan software to identify and analyze WWTPs located within and close to spatiotemporal clusters of legionellosis detected in Quebec between 2016 and 2020. In parallel, WWTPs were ranked according to their pollutant load, flow rate and treatment type. These parameters were used to evaluate the WWTP susceptibility to generate and disperse bioaerosols. Results show that 37 of the 874 WWTPs are located inside a legionellosis cluster study zone, including six of the 40 WWTPs ranked most susceptible. In addition, two susceptible WWTPs located within an extended area of 2.5 km from the study zone (2.5-km buffer) were included, for a total of 39 WWTPs. The selected 39 WWTPs were further studied to document proximity of population, dominant wind direction, and surrounding water quality. Samples collected from the influent and the effluent of six selected WWTPs revealed the presence of Legionella spp. in 92.3% of the samples. Lp and Lp serogroupg 1 (Lp sg1) were detected below the limit of quantification in 69% and 46% of the samples, respectively. The presence of Legionella in wastewater and the novel statistical approach presented here provides information to the public health authorities regarding the investigation of WWTPs as a possible source of Legionella exposure, sporadic cases, and clusters of legionellosis.

Experimental evaluation of a full-scale in-duct UV germicidal irradiation system for bioaerosols inactivation.

Bioaerosols control techniques, especially ultraviolet germicidal irradiation (UVGI) are gaining attention due to increasing needs for controlling of health risk caused by airborne biocontaminants. The effectiveness of a full-scale in-duct UVGI air disinfection system was investigated. One bacterium, a wild type Escherichia coli, and three fungal spores, Penicillium aragonense, Rhodotorula glutinis, and Cladosporium sp., were selected as test organisms and their inactivation under different conditions representative of a real application in HVAC systems were investigated. The results demonstrated that inactivation of airborne E. coli by the UVGI system was extremely effective, with >99.5 % of the input E. coli inactivated at a residence time lower than 0.36 s in the disinfection section. Airborne fungal spores were less susceptible to UV irradiation than E. coli. Under same conditions, viable counts reduction of P. aragonense, R. glutinis, and Cladosporium sp. spores were 53 %, 63 % and 73 %, respectively. The effect of UV light intensity, air flowrate and relative humidity were analyzed separately. A simplified model based on redefinition of the parameters in the classical inactivation kinetic equation was used to simulate the inactivation of airborne contaminants in the in-duct system under different conditions. The results showed that the simplified model was adequate to estimate disinfection efficacy of different bioaerosols by the UVGI system which could be useful for system design. Overall, this study shows that such in-duct UVGI systems can provide significant control of bioaerosols.

Airborne Influenza Virus in Daycare Centers.

In this study, we investigated the concentration of airborne influenza virus in daycare centers and influencing factors, such as common cold prevalence, air pollutants, and meteorological factors. A total of 209 air samples were collected from daycare centers in Kaohsiung and the influenza virus was analyzed using real-time quantitative polymerase chain reaction. Air pollutants and metrological factors were measured using real-time monitoring equipment. Winter had the highest positive rates of airborne influenza virus and the highest prevalence of the common cold, followed by summer and autumn. The concentration of CO was significantly positively correlated with airborne influenza virus. Daycare center A, with natural ventilation and air condition systems, had a higher concentration of airborne influenza A virus, airborne fungi, and airborne bacteria, as well as a higher prevalence of the common cold, than daycare center B, with a mechanical ventilation system and air purifiers, while the concentrations of CO2, CO, and UFPs in daycare center A were lower than those in daycare center B. We successfully detected airborne influenza virus in daycare centers, demonstrating that aerosol sampling for influenza can provide novel epidemiological insights and inform the management of influenza in daycare centers.

Alteration of the health effects of bioaerosols by chemical modification in the atmosphere: A review.

Bioaerosols are a subset of important airborne particulates that present a substantial human health hazard due to their allergenicity and infectivity. Chemical reactions in atmospheric processes can significantly influence the health hazard presented by bioaerosols; however, few studies have summarized such alterations to bioaerosols and the mechanisms involved. In this paper, we systematically review the chemical modifications of bioaerosols and the impact on their health effects, mainly focusing on the exacerbation of allergic diseases such as asthma, rhinitis, and bronchitis. Oxidation, nitration, and oligomerization induced by hydroxyl radicals, ozone, and nitrogen dioxide are the major chemical modifications affecting bioaerosols, all of which can aggravate allergenicity mainly through immunoglobulin E pathways. Such processes can even interact with climate change including the greenhouse effect, suggesting the importance of bioaerosols in the future implementation of carbon neutralization strategies. In summary, the chemical modification of bioaerosols and the subsequent impact on health hazards indicate that the combined management of both chemical and biological components is required to mitigate the health hazards of particulate air pollution.

Filling the knowledge gap: Scoping review regarding sampling methods, assays, and further requirements to assess airborne viruses.

Assessment of occupational exposure to viruses is crucial to identify virus reservoirs and sources of dissemination at an early stage and to help prevent spread between employees and to the general population. Measuring workers' exposure can facilitate assessment of the effectiveness of protective and mitigation measures in place. The aim of this scoping review is to give an overview of available methods and those already implemented for airborne virus' exposure assessment in different occupational and indoor environments. The results retrieved from the different studies may contribute to the setting of future standards and guidelines to ensure a reliable risk characterization in the occupational environments crucial for the implementation of effective control measures. The search aimed at selecting studies between January 1st 2010 and June 30th 2023 in the selected databases. Fifty papers on virus exposure assessment fitted the eligibility criteria and were selected for data extraction. Overall, this study identified gaps in knowledge regarding virus assessment and pinpointed the needs for further research. Several discrepancies were found (transport temperatures, elution steps, …), as well as a lack of publication of important data related to the exposure conditions (contextual information). With the available information, it is impossible to compare results between studies employing different methods, and even if the same methods are used, different conclusions/recommendations based on the expert judgment have been reported due to the lack of consensus in the contextual information retrieved and/or data interpretation. Future research on the field targeting sampling methods and in the laboratory regarding the assays to employ should be developed bearing in mind the different goals of the assessment.

Atmospheric dispersal shapes rapid bacterial colonization of Icelandic Lava Rocks.

Microorganisms released into the atmosphere by various disturbances can travel significant distances before depositing, yet their impact on community assembly remains unclear. To address this, we examined atmospheric and lithospheric bacterial communities in 179 samples collected at two distinct Icelandic volcanic sites: a small volcanic island Surtsey, and a volcanic highland Fimmvörðuháls using 16S rRNA amplicon sequencing. Airborne microbial communities were similar between sites while significant differences emerged in the communities on lava rocks after 1-year exposure. SourceTracker analysis revealed distinct bacterial populations in the atmosphere and the lava rocks with surrounding soil contributed more significantly to lava rock microbial composition. Nevertheless, shared genera among air, rocks, and local sources, suggested potential exchange between these environments. The prevalent genera shared between rocks and potential sources exhibited stress-resistant properties, likely helping their survival during air transportation and facilitating their colonization of the rocks. We hypothesize that the atmosphere serves as a conduit for locally sourced microbes and stress-resistant distant-sourced microbes. Additionally, bacterial communities on the lava rocks of Fimmvörðuháls showed remarkable similarity after 1 and 9 years of exposure, suggesting rapid establishment. Our study reveals that atmospheric deposition significantly influences bacterial community formation, potentially influencing ecosystem dynamics and microbial communities' resilience.

Bioaerosol Exposures and Respiratory Diseases in Cannabis Workers.

PURPOSE OF REVIEW: This review investigates occupational inhalation hazards associated with biologically derived airborne particles (bioaerosols) generated in indoor cannabis cultivation and manufacturing facilities. RECENT FINDINGS: Indoor cannabis production is growing across the US as are recent reports of respiratory diseases among cannabis workers, including occupational asthma morbidity and mortality. More information is needed to understand how bioaerosol exposure in cannabis facilities impacts worker health and occupational disease risk. Preliminary studies demonstrate a significant fraction of airborne particles in cannabis facilities are comprised of fungal spores, bacteria, and plant material, which may also contain hazardous microbial metabolites and allergens. These bioaerosols may pose pathogenic, allergenic, toxigenic, and pro-inflammatory risks to workers. The absence of multi-level, holistic bioaerosol research in cannabis work environments necessitates further characterization of the potential respiratory hazards and effective risk prevention methods to safeguard occupational health as the cannabis industry continues to expand across the US and beyond.

Spatial distribution of bioaerosols and evaluation of four ventilation method on controlling their diffusion in a typical enhanced biosafety level 2 laboratory.

Biosafety laboratories are critical in many fields. However, experimenters associated the infection risk from biological aerosols. In this study, by conducting experiments on the release and collection of bioaerosols within a typical BSL-2 + laboratory, the spatial distribution of bioaerosols was tracked. Numerical calculations were employed to obtain and visualize the airflow patterns and aerosol dispersion paths of four ventilation methods. The results indicated that equipment and tables led to uneven airflow distribution within the laboratory. The comparison results of the four evaluation indicators showed that the air age distribution of UU (Upward supply and upward return) mode and CD (Cross-supply and downward return) mode was superior, with air change efficiency values of 0.595 and 0.603, respectively. Additionally, the contaminant removal index of CD mode was 1.48, significantly higher than the other ventilation methods. The statistical results of the contaminant dispersion index also indicated that CD mode was most conducive to diluting aerosols in the spatial environment. The LD (lateral supply and downward return) mode may lead to airflow short-circuiting. The UD (upward supply and downward return) mode can provide balanced protection for laboratory. Overall, CD mode performed the best among the four ventilation methods, followed by UU mode.

Influence of CO2 and Dust on the Survival of Non-Resistant and Multi-Resistant Airborne E. coli Strains.

The airborne transmission of bacterial pathogens poses a significant challenge to public health, especially with the emergence of antibiotic-resistant strains. This study investigated environmental factors influencing the survival of airborne bacteria, focusing on the effects of different carbon dioxide (CO2) and dust concentrations. The experiments were conducted in an atmospheric simulation chamber using the non-resistant wild-type E. coli K12 (JM109) and a multi-resistant variant (JM109-pEC958). Different CO2 (100 ppm, 800 ppm, 3000 ppm) and dust concentrations (250 µg m-3, 500 µg m-3, 2000 µg m-3) were tested to encompass a wide range of CO2 and dust levels. The results revealed that JM109-pEC958 exhibited greater resilience to high CO2 and dust concentrations compared to its non-resistant counterpart. At 3000 ppm CO2, the survival rate of JM109 was significantly reduced, while the survival rate of JM109-pEC958 remained unaffected. At the dust concentration of 250 µg m-3, JM109 exhibited significantly reduced survival, whereas JM109-pEC958 did not. When the dust concentration was increased to 500 and 2000 µg m-3, even the JM109-pEC958 experienced substantially reduced survival rates, which were still significantly higher than those of its non-resistant counterpart at these concentrations. These findings suggest that multi-resistant E. coli strains possess mechanisms enabling them to endure extreme environmental conditions better than non-resistant strains, potentially involving regulatory genes or efflux pumps. The study underscores the importance of understanding bacterial adaptation strategies to develop effective mitigation approaches against antibiotic-resistant bacteria in atmospheric environments. Overall, this study provides valuable insights into the interplay between environmental stressors and bacterial survival, serving as a foundational step towards elucidating the adaptation mechanisms of multi-resistant bacteria and informing strategies for combating antibiotic resistance in the atmosphere.

Aerosol soluble proteins in Asian dust in southwestern Japan.

Aerosol proteins, as core biological components of bioaerosols, are garnering increasing attention due to their environmental significance, including their roles in atmospheric processes and associated health risks. However, observational data on the proteins are very limited, leaving their distribution and variation in the atmosphere poorly understood. To investigate the long-distance transport of proteins with Asian dust in the Northern Hemisphere middle latitude westerlies to remote downwind areas, we quantified the soluble proteins in aerosol particles, referred to as aerosol soluble proteins (ASPs), collected in the coastal city of Kumamoto, Japan, during the spring of 2023, when three dust events occurred. The concentration of ASPs ranged from 0.22 to 1.68 µg m-3, with an average concentration of 0.73 ± 0.36 µg m-3 under dust conditions and 0.31 ± 0.05 µg m-3 under non-dust conditions. During the dust periods, the largest concentration of ASPs (1.68 µg m-3) coincided with the peak concentration of suspended particulate matter, and the concentration strongly correlated with the mass concentration of particles larger than 2.5 µm, indicating a close dependence of ASPs on dust particles. Primary estimations indicated a dry deposition flux of ASPs at approximately 1.10 ± 0.87 mg m-2 d-1 under the dust conditions. These results prove that Asian dust efficiently transports proteins, facilitating their dispersion in the atmosphere.

Spectroscopic detection of bioaerosols with the wibs-4+: Anthropogenic and meteorological impacts.

This research builds upon a previous study that explored the potential of the modified WIBS-4+ to selectively differentiate and detect different bioaerosol classes. The current work evaluates the influence of meteorological and air quality parameters on bioaerosol concentrations, specifically pollen and fungal spore dynamics. Temperature was found to be the most influential parameter in terms of pollen production and release, showing a strong positive correlation. Wind data analysis provided insights into the potential geographic origins of pollen and fungal spore concentrations. Fungal spores were primarily shown to originate from a westerly direction, corresponding to agricultural land use, whereas pollen largely originated from a North-easterly direction, corresponding to several forests. The influence of air quality was also analysed to understand its potential impact on the WIBS fluorescent parameters investigated. Most parameters had a negative association with fungal spore concentrations, whereas several anthropogenic influences showed notable positive correlations with daily pollen concentrations. This is attributed to similar driving forces (meteorological parameters) and geographical origins. In addition, the WIBS showed a significant correlation with anthropogenic pollutants originating from combustion sources, suggesting the potential for such modified spectroscopic instruments to be utilized as air quality monitors. By combining all meteorological and pollution data along with WIBS-4+ channel data, a set of Multiple Linear Regression (MLR) analyses were completed. Successful results with R2 values ranging from 0.6 to 0.8 were recorded. The inclusion of meteorological parameters was dependent on the spore or pollen type being examined.

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.

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.

Tracking bioaerosol exposure among municipal solid waste workers using hematological and inflammatory biomarkers.

High levels of bioaerosols may exist in the air of municipal solid waste (MSW) management facilities, constituting a significant occupational hazard for workers. In this study, we investigated the potential association between exposure to bioaerosols and inflammatory biomarkers among municipal solid waste workers (MSWWs) at both the landfill site and the municipal solid waste transfer station (MSWTS), in comparison to a control group without exposure. Air sampling was conducted at six points around the landfill, two points at the MSWTS, and one point in a public park (as a control area) during the spring and summer of 2019. The results of our study revealed that airborne pathogens were highly prevalent at the sampling points, especially in the active zone and leachate collection pond. Aspergillus species were the predominant fungal species detected in this study, with the highest occurrence observed for Aspergillus flavus (83.3%), Aspergillus niger, and Aspergillus fumigatus (75.0%). Furthermore, Staphylococcus species accounted for over 75% of the total bacterial bioaerosols detected across all study areas. The blood test results of workers revealed a significant increase in platelets (PLT), immunoglobulin G (IgG), white blood cells (WBC), neutrophils, basophils, and high-sensitivity C-reactive protein (hs-CRP) compared to the control group. Conversely, platelet distribution width (PDW), mean platelet volume (MPV), and platelet-large cell ratio (P-LCR) in the exposed subjects exhibited a decreasing trend compared to the control group. These findings suggest a potential association between exposure to bioaerosols, particularly in the vicinity of open dumpsites, and elevated levels of hematologic and inflammatory markers in circulation. Furthermore, the influence of smoking status and confounding factors appears to be significant in both the control and exposure groups.

The atmosphere: a transport medium or an active microbial ecosystem?

The atmosphere may be Earth's largest microbial ecosystem. It is connected to all of Earth's surface ecosystems and plays an important role in microbial dispersal on local to global scales. Despite this grand scale, surprisingly little is understood about the atmosphere itself as a habitat. A key question remains unresolved: does the atmosphere simply transport microorganisms from one location to another, or does it harbour adapted, resident, and active microbial communities that overcome the physiological stressors and selection pressures the atmosphere poses to life? Advances in extreme microbiology and astrobiology continue to push our understanding of the limits of life towards ever greater extremes of temperature, pressure, salinity, irradiance, pH, and water availability. Earth's atmosphere stands as a challenging, but potentially surmountable, extreme environment to harbour living, active, resident microorganisms. Here, we confront the current understanding of the atmosphere as a microbial habitat, highlighting key advances and limitations. We pose major ecological and mechanistic questions about microbial life in the atmosphere that remain unresolved and frame the problems and technical pitfalls that have largely hindered recent developments in this space, providing evidence-based insights to drive future research in this field. New innovations supported by rigorous technical standards are needed to enable progress in understanding atmospheric microorganisms and their influence on global processes of weather, climate, nutrient cycling, biodiversity, and microbial connectivity, especially in the context of rapid global change.

Needles in haystacks: monitoring the potential escape of bioaerosolised antibacterial resistance genes from wastewater treatment plants with air and phyllosphere sampling.

Wastewater treatment plants are well-known point sources of emissions of antibacterial resistance genes (ARGs) into the environment. Although most work to date has focused on ARG dispersal via effluent, aerial dispersal in bioaerosols is a poorly understood, but likely important vector for ARG dispersal. Recent evidence suggests that ARG profiles of the conifer needle phyllosphere could be used to measure bioaerosol dispersal from anthropogenic sources. Here, we assessed airborne dispersal of ARGs from wastewater treatment plants in Wales, UK and Quebec, Canada, using conifer needles as passive bioaerosol monitors. ARG profiles of wastewater were compared to those of conifer phyllosphere using high-throughput qPCR. ARG richness was significantly lower in conifer phyllosphere samples than wastewater samples, though no differences were observed across the dispersal gradients. Mean copy number of ARGs followed a similar trend. ARG profiles showed limited, but consistent patterns with increasing distance from wastewater treatment plants, but these did not align with those of wastewater samples. For example, proportional abundance of aminoglycosides decreased over the dispersal gradient in Wales, whereas mobile genetic elements showed the inverse relationship. In summary, while distinct ARG profiles exist along dispersal gradients, links to those of wastewater were not apparent.

Comparison of Three Air Sampling Methods for the Quantification of Salmonella, Shiga-toxigenic Escherichia coli (STEC), Coliforms, and Generic E. coli from Bioaerosols of Cattle and Poultry Farms.

Recent fresh produce outbreaks potentially associated with bioaerosol contamination from animal operations in adjacent land highlighted the need for further study to better understand the associated risk. The purpose of this research was to evaluate three sampling methods for quantifying target bacterial bioaerosols from animal operations. A dairy cattle and poultry farm located in Georgia, U.S. were visited six times each. Air was collected for 10 min using: 2-stage Andersen impactor with and without mineral oil overlay and impingement samplers. Sampling devices were run concurrently at 0.1, 1, and 2 m heights (n = 36). Andersen samplers were loaded with CHROMagar™ Salmonella, CHROMagar™ STEC, or Brilliance™ coliforms/E. coli. The impingement sampler contained buffered peptone water (20 mL) which was vacuum filtered through a 0.45 µm filter and placed onto the respective media. Plates were incubated at 37 ℃ for 48 h. PCR confirmation followed targeting ttr for Salmonella and stx1, stx2, and eae genes for STEC. No significant differences were found among methods to quantify coliforms and E. coli. Salmonella and STEC bioaerosols were not detected by any of the methods (Limit of detection: 0.55 log CFU/m3). E. coli bioaerosols were significantly greater in the poultry (2.76-5.00 log CFU/m3) than in the cattle farm (0.55-2.82 log CFU/m3) (p < 0.05), and similarly distributed at both stages in the Andersen sampler (stage 1:>7 µm; stage 2: 0.65-7 µm particle size). Sampling day did not have a significant effect on the recovery of coliforms/E. coli bioaerosols in the poultry farm when samples were taken at the broiler house exhaust fan (p > 0.05). A greater and constant emission of coliforms and E. coli bioaerosols from the poultry farm warrants further investigation. These data will help inform bioaerosol sampling techniques which can be used for the quantification of bacterial foodborne pathogens and indicator organisms for future research.