Bioaerosol Sampling Research Articles

Optimisation of bioaerosol sampling using an ultralight aircraft: A novel approach in determining the 3-D atmospheric biodiversity

Bioaerosols, such as pollen and fungal spores, are routinely monitored for agricultural, medical or urban greening practices, but sampling methodology is largely relying on techniques more than half a century old. Moreover, biomonitoring campaigns often take place in urban environments, although sources can be located outside cities’ borders with ampler vegetation. Therefore, the question arises whether we are accurately picturing the biodiversity and abundance of regional bioaerosols and whether those locally detected might derive from long-distance transport, horizontally or vertically. To answer the above, we used novel, mobile monitoring devices, and aerial measurement units, like aircrafts, so as to explore bioaerosol concentrations at a variety of altitudes.An ultralight aircraft was equipped with a sampling device for bioaerosols. The device consisted of duplicate isokinetic impactors that match the physical functioning and the microscopic quantification method of the widely used ground-based Hirst-type impactors. Isokinetic airflow was realized by adjusting the air flux at the impactors’ inlet to the airspeed of the aircraft. Three campaigns were made, where the comparability, efficiency and accuracy of different sampling devices were determined, namely of the abovementioned impactor, and of the mobile conventional Hirst-type pollen sampler. The campaigns involved measurements from ground level (0 m altitude) up to 900 m (above ground level (agl)) via flights.Our results showed that aircraft-based airborne pollen concentration measurements were consistently higher than those of all other devices, regardless of the altitude and sampling time. It is noteworthy that the pollen concentration exceeded 500 pollen grains/m3 at >900 m of altitude, this concentration being 1.77 times higher than that simultaneously measured at ground level. Likewise, the diversity of pollen was also higher at higher altitude.Our results indicate the usability and superiority of small aircraft and high-flow impactors for research, achieving higher biodiversity and abundance over a shorter sampling interval compared to conventional volumetric techniques. Higher pollen amounts at higher altitudes also point at the necessity to monitor bioaerosols across the vertical dimension, especially in densely populated areas and high-traffic air space.

SARS-CoV-2 infection and transmission via the skin to oro-nasal route with the production of bioaerosols in the ferret model.

Direct and indirect transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been attributed to virus survival in droplets, bioaerosols and on fomites including skin and surfaces. Survival of SARS-CoV-2 variants of concern (Alpha, Beta, Gamma, and Delta) on the skin and virus transference following rounds of skin-to-skin contact were assessed on porcine skin as a surrogate for human skin. SARS-CoV-2 variants were detectable on skin by RT-qPCR after 72 h at biologically relevant temperatures (35.2 °C) with viral RNA (vRNA) detected after ten successive skin-to-skin contacts. Skin-to-skin virus transmission to establish infection in ferrets as a model for mild/asymptomatic SARS-CoV-2 infection in mustelids and humans was also investigated and compared to intranasal ferret inoculation. Naïve ferrets exposed to Delta variant SARS-CoV-2 in a 'wet' or 'dry' form on porcine skin resulted in robust infection with shedding detectable for up to 14 days post-exposure, at comparable viral loads to ferrets inoculated intranasally. Transmission of SARS-CoV-2 to naïve ferrets in direct contact with infected ferrets was achieved, with environmental contamination detected from ferret fur swabs and air samples. Genetic substitutions were identified in bioaerosol samples acquired following single contact passage in ferrets, including Spike, ORF1ab, and ORF3a protein sequences, suggesting a utility for monitoring host adaptation and virus evolution via air sampling. The longevity of SARS-CoV-2 variants survival directly on the skin and skin-to-skin transference, enabling subsequent infection via the skin to oro-nasal contact route, could represent a pathway for SARS-CoV-2 infection with implications to public and veterinary health.

Bioaerosol sampling and bioanalysis: Applicability of the next generation impactor for quantifying Legionella pneumophila in droplet aerosols by flow cytometry

Bioaerosol generation, sampling, and cultivation-independent quantification of pathogenic bacteria play a crucial role in studying dose-response effects of Legionella pneumophila. Here, the Next Generation Impactor (NGI), initially created for pharmaceutical inhaling studies, was assessed for its potential to sample airborne bioaerosols and to separate size-dependent wet droplets by incrementally increasing the airflow speed. This stainless-steel sampler was shown in this study to be suitable for sampling prior to cultivation-independent analysis of pathogen-containing bioaerosols using washable cups. The applicability was studied by quantifying the total and intact cell count of L. pneumophila by flow cytometry after being dispersed into a droplet aerosol. Our results demonstrate a high total sampling efficiency of 95.5% ± 11.8% despite a lower biological sampling efficiency of 59.7% ± 16.5% for dry aerosols. However, by elevating the relative humidity (RH) to 100% in a liquid aerosolization unit, the biological sampling efficiency increased to over 90% for L. pneumophila. More than 50% of the cells were found in stage 1 using the liquid aerosolization unit. In comparison, 80% of the cells were sampled in stages 4–6 at 30% RH. Specifically, while at 100% RH, the droplet size mattered, at 30% RH, the size distribution of dry particles, in this case L. pneumophila, was relevant due to evaporation processes, which explains the size differences. These findings indicate the potential of the NGI for further exploration and application in studying other aerosol-borne pathogens, especially concerning the size distribution of wet droplets, viability, or effect-based bioanalysis.

Development of a Novel Electrostatic-Based Bioaerosol Sampler.

On-site bioaerosol monitoring is essential for estimating microbial biomass and mitigating the risk of infection induced by aerosol transmission. This study introduces a novel electrostatic bioaerosol sampler, which is fabricated by the use of 3D printing, for rapid bioaerosol collection. Aerosol particles were charged and enriched in the sampler. Relationships between particle sizes and collection efficiencies under varying charging voltages were established using a charging model. The design of the sampler was optimized using commercial software, incorporating electrostatic field analysis, computational fluid dynamics (CFD), and particle trajectory simulations. To validate the sampler's collection efficiency, polystyrene (PS) spheres in an aerosol dispenser were atomized into an aerosol. The sampler collection efficiency exceeded 90% for particles larger than 1.2 μm under an applied voltage of 4.7 kV and an airflow rate of 2 L/min. The enrichment capacity was greater than 153,000 for particles larger than 1.2 μm under an applied voltage of 4.7 kV and an airflow rate of 8 L/min. With the merits of low cost, miniaturization, and high collection efficiency, the sampler can be used to collect samples on-site and in remote areas to verify the pathogens and reduce the risk of infection through aerosol transmission.

Optimal environmental sampling conditions for electrostatic aerosol-to-hydrosol collection of airborne viruses

Due to adverse effects of viral outbreaks on human health, accurate detection of airborne pathogens is essential. Among many methods available for bioaerosol sampling, electrostatic precipitation (ESP) has been used to directly collect bioaerosols as hydrosols. The performance of an ESP sampler depends on its design, operational and environmental parameters such as air relative humidity (RH), air temperature, sampling liquid type and liquid temperature. Thus, it is essential to identify and maintain optimal conditions throughout sampling process to operate the sampler at its highest capacity. This study provides crucial insights into parameters that affect the collection efficiency of the aerosol-to-hydrosol ESP sampler and its virus recovery. The results indicate that air temperature does not affect collection efficiency, meanwhile, air RH, sampling liquid temperature, and salt concentration are the main parameters that significantly affect collection efficiency. Likewise, when deionized water is used as sampling liquid, hydrogen peroxide concentration increases proportionally with increasing air RH, resulting in significant decrease of virus viability. Consequently, for ESP samplers similar to our study, the following conditions are recommended: air RH of 55–65%, air and sampling liquid temperature of 37 °C, and a mixture of 10–20 mM ascorbic acid in PBS as sampling liquid.

Microfluidics-based condensation bioaerosol sampler for multipoint airborne virus monitoring

To facilitate rapid monitoring of airborne viruses, they must be collected with high efficiency and concentrated in a small volume of a liquid sample. In addition, the development of low-cost miniaturized samplers is essential for multipoint monitoring. Thus, in an attempt to fulfill these requirements, this study developed a microfluidic condensation bioaerosol sampler (MCBS). The developed sampler comprised two parts: a virus growth section and a virus droplet-to-liquid sample conversion section, each of which was fabricated on a chip using microfluidic technology. The condensation nucleus growth technique used in the virus growth section grew nanometer-sized airborne viruses into micro-sized droplets, making it possible to collection of viruses easier and with high efficiency. In addition, the virus droplet-to-liquid sample conversion section controlled the transport of droplets based on electrowetting technology. This enabled the collected airborne viruses to be concentrated in tens of microliters of the liquid sample. To evaluate the performance of both the sections, the virus dropletization, virus collection efficiency, and virus droplet-to-liquid sample conversion efficiency were evaluated through quantitative experiments. H1N1 and HCOV-229E viruses were used to conduct quantitative experiments on MCBS. We could obtain virus liquid samples with at 72.8- and 89.9-times higher concentration through 1:1 evaluation with a commercial sampler. Thus, the developed sampler facilitated efficient collection and concentration of airborne viruses in a compact, cost-effective manner. This is expected to facilitate rapid and accurate multipoint monitoring of viral aerosols.

Concentrating viable airborne pathogens using a virtual impactor with a compact water-based condensation air sampler.

Pathogens can be collected from air and detected in samples by many methods. However, merely detecting pathogens does not answer whether they can spread disease. To fully assess health risks from exposure to airborne pathogens, the infectivity of those agents must be assessed. Air samplers which operate by growing particles through water vapor condensation and subsequently collecting them into a liquid medium have proven effective at conserving the viability of microorganisms. We present a study that assessed performance improvement of one such sampler, BioSpot-GEM™, gained by augmenting it with an upstream virtual impactor (VI) designed to concentrate particles in aerosols. We demonstrate that such an integrated unit improved the collection of live Escherichia coli by a median Concentration Factor (C F ) of 1.59 and increased the recovery of viable human coronavirus OC43 (OC43) by a median C F of 12.7 as compared to the sampler without the VI. Our results also show that OC43 can be concentrated in this way without significant loss of infectivity. We further present that the small BioSpot-GEM™ bioaerosol sampler can collect live E. coli at an efficiency comparable to the larger BioSpot-VIVAS™ bioaerosol sampler. Our analyses show potential benefits toward improving the collection of viable pathogens from the air using a more portable water-based condensation air sampler while also highlighting challenges associated with using a VI with concentrated bioaerosols. This work can aid further investigation of VI usage to improve the collection of pathogens from air ultimately to better characterize health risks associated with airborne pathogen exposures.

First observations on airborne prokaryotes in a subArctic Atlantic marine area

Among extreme environments, bioaerosol includes a wide range of primary atmospheric organic particles associated with and emitted by living and dead organisms. Bioaerosol samples were collected along two transects at a subArctic Atlantic spatial scale, including the eastern Fram Strait and the Greenland, Norwegian, and North Seas. This study was aimed at first estimating microscopically the prokaryotic abundance, biomass and phenotypic traits, along with the number of potential viable and respiring cells. Moreover, physiological profiles at community level were assessed. Prokaryotic abundance ranged from 104 to 107 cells m−3, with the predominance of small sized cells (0.1 μm3). Prokaryotic biomass reached higher values (mean value 233 μg C m−3) in relation to the occurrence of large sized rods. Overall, the percentage of the viable cells was lower than the dead ones, while respiring cells were in lower abundance than total cells. The physiological profiles revealed various potential metabolic pathways among the samples, highlighting the utilization of phosphate-carbon, carboxylic and amino acids. These first results on the metabolism and physiology of microbes, which survived transport in the atmosphere of the Northern Hemisphere, suggest that bioaerosol constitutes an extremely dynamic environment of remarkable ecological interest, also considering future global warming scenarios.

High Prevalence of Highly Pathogenic Avian Influenza: A Virus in Vietnam's Live Bird Markets.

In recent years, Vietnam has suffered multiple epizootics of influenza in poultry. From 10 January 2019 to 26 April 2021, we employed a One Health influenza surveillance approach at live bird markets (LBMs) and swine farms in Northern Vietnam. When the COVID-19 pandemic permitted, each month, field teams collected oral secretion samples from poultry and pigs, animal facility bioaerosol and fecal samples, and animal worker nasal washes at 4 LBMs and 5 swine farms across 5 sites. Initially samples were screened with molecular assays followed by culture in embryonated eggs (poultry swabs) or Madin-Darby canine kidney cells (human or swine swabs). Many of the 3493 samples collected had either molecular or culture evidence for influenza A virus, including 314 (37.5%) of the 837 poultry oropharyngeal swabs, 144 (25.1%) of the 574 bioaerosol samples, 438 (34.9%) of the 1257 poultry fecal swab samples, and 16 (1.9%) of the 828 human nasal washes. Culturing poultry samples yielded 454 influenza A isolates, 83 of which were H5, and 70 (84.3%) of these were highly pathogenic. Additionally, a positive human sample had a H9N2 avian-like PB1 gene. In contrast, the prevalence of influenza A in the swine farms was much lower with only 6 (0.4%) of the 1700 total swine farm samples studied, having molecular evidence for influenza A virus. This study suggests that Vietnam's LBMs continue to harbor high prevalences of avian influenza A viruses, including many highly pathogenic H5N6 strains, which will continue to threaten poultry and humans.

Assessment of the partial partition effect on the control of bioaerosol transmission in a one bed ward

The partition is a boundary that can effectively suppress the spread of bioaerosols. It has been widely used in public places such as restaurants and shopping malls during COVID-19 pandemic. Inspired by this, this study explored the effect of partial partitions in limiting the diffusion of bioaerosols in a typical isolation ward. In bioaerosol experiments, Serratia marcescens was used as bioaerosol releasing material to study the bioaerosol distribution. Simultaneously, the result of bioaerosol sampling was used to validate the numerical model. Furthermore, the effect of five partitions strategies on aerosol diffusion was further explore by Computational fluid dynamics. The flow patterns in the ward and the spatiotemporal distribution of aerosols were obtained and analyzed. The results showed that the best strategy could increase bioaerosol removal effectiveness index from 0.569 to 0.696, which accelerated the bioaerosol removal. Nevertheless, only selecting a reasonable size and installation position, can the partial partition play a positive role in the interruption of aerosol diffusion. Moreover, small-area partial partition can achieve efficient control of bioaerosol diffusion as well.

Identification of antibiotic-resistant, gram-negative bacteria in sewage and bioaerosols from a wastewater treatment plant: A genotypic and phenotypic study

ABSTRACT Antimicrobial resistance (AMR) is a growing global public health concern. This study aimed to investigate the presence of bacteria and antibiotic resistance genes in the environment, more specifically in sewage and bioaerosol samples collected at a Wastewater Treatment Plant (WWTP). Bacterial species were identified and tested for antibiotic sensitivity. In addition, bla CTX-M, bla SHV, bla TEM and bla KPC genotypes, including those related to resistance to carbapenems, were detected by PCR. The results of this research are extremely important for understanding the mechanisms involved in antimicrobial resistance and for preventing the dissemination of these antibiotic-resistant microorganisms. This study contributes to the identification of sources of antibiotic resistance in the environment and implementation of AMR control and prevention strategies while preserving the effectiveness of antibiotics and public health.

Impact of cultivating different Ocimum species on bioaerosol bacterial communities and functional genome at an agricultural site

The effects of the surrounding environment on the bacterial composition of bioaerosol were well documented for polluted and contaminated sites. However, there is limited data on the impact of plant species, especially those that produce aromas, on bioaerosol composition at agricultural sites. Hence, the aim of this study is to evaluate the variability in bacterial communities present in bioaerosol samples collected from agricultural sites with aroma-producing crops. For this, PM2.5, PM10, and bioaerosol samples were collected from agricultural fields growing Ocimum [two varieties of O. sanctum (CIM-Aayu and CIM-Angana)] and O. kilimandscharicum (Kapoor), nearby traffic junctions and suburban areas. PM2.5 and PM10 concentrations at the agricultural site were in between the other two polluted sites. However, bioaerosol concentration was lower at agricultural sites than at other sites. The culturable bacteria Bacillus subtilis, Bacillus tequilensis, and Staphylococcus saprophyticus were more prevalent in agricultural sites than in other areas. However, the composition of non-culturable bacteria varied between sites and differed in three fields where Ocimum was cultivated. The CIM-Aayu cultivated area showed a high bacterial richness, lower Simpson and Shannon indices, and a distinctive metabolic profile. The sites CIM-Angana and CIM-Kapoor had a higher abundance of Aeromonas, while Pantoea and Pseudomonas were present at CIM-Aayu. Acinetobacter, Staphylococcus, and Bacillus were the dominant genera at the other two sites. Metabolic profiling showed that the CIM-Aayu site had a higher prevalence of pathways related to amino acid and carbohydrate metabolism and environmental information processing compared to other sites. The composition of bioaerosol among the three different Ocimum sites could be due to variations in the plant volatile and cross-feeding nature of bacterial isolates, which further needs to be explored.

Prevalence and antibiotic susceptibility patterns of Escherichia coli and Staphylococcus aureus isolated from bioaerosols of local markets located in Agulu, Anambara State

There is a need for improved understanding of aerosol-acquired disease due to the increasing environmental load of antibiotic resistant bacteria. Multidrug-resistant Escherichia coli and Staphylococcus aureus are examples of bacteria for which the role of exposure to aerosolized organisms in disease transmission should be more closely evaluated globally. This study is aimed at evaluating the prevalence and antibiogram of S. aureus and E. coli present in market bioaerosols, located in Nwagu-Agulu, Anambra Nigeria. Impaction sample collection method was used to collect bioaerosol samples within the market using nutrient agar as the collection media, A combination of the cultural and microscopic features as well as selected biochemical tests were employed to identify the isolates. The Kirby-Bauer disk diffusion method was used to determine antibiotic resistance to selected commonly used antibiotics. A total of 76 S. aureus and 24 E. coli were isolated. Both organisms have been implicated in community-acquired infections. The most common bacteria isolated were Staphylococcus aureus. Varying levels of resistance were observed by these isolates to selected commonly use antibiotics. There is high prevalence of S. aureus and E. coli within Nwagu market Agulu, Anambra State, Eastern region of Nigeria. The organisms have very high rates of resistance to antibiotics (MDR) as reported by other workers. The government as well as Health professionals must play an important role in the prevention of community acquired infections via regular environmental monitoring of open markets bioaerosol and establishing a common permissible limits or concentration for bioaerosols within the markets.

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.

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.

Bioaerosol Sampling Devices and Pretreatment for Bacterial Characterization: Theoretical Differences and a Field Experience in a Wastewater Treatment Plant.

Studies on bioaerosol bacterial biodiversity have relevance in both ecological and health contexts, and molecular methods, such as 16S rRNA gene-based barcoded sequencing, provide efficient tools for the analysis of airborne bacterial communities. Standardized methods for sampling and analysis of bioaerosol DNA are lacking, thus hampering the comparison of results from studies implementing different devices and procedures. Three samplers that use gelatin filtration, swirling aerosol collection, and condensation growth tubes for collecting bioaerosol at an aeration tank of a wastewater treatment plant in Trieste (Italy) were used to determine the bacterial biodiversity. Wastewater samples were collected directly from the untreated sewage to obtain a true representation of the microbiological community present in the plant. Different samplers and collection media provide an indication of the different grades of biodiversity, with condensation growth tubes and DNA/RNA shieldTM capturing the richer bacterial genera. Overall, in terms of relative abundance, the air samples have a lower number of bacterial genera (64 OTUs) than the wastewater ones (75 OTUs). Using the metabarcoding approach to aerosol samples, we provide the first preliminary step toward the understanding of a significant diversity between different air sampling systems, enabling the scientific community to orient research towards the most informative sampling strategy.

Comparison of the density and composition of bacteria and fungi in the air of anoxic and aeration tanks in the wastewater treatment plant

The bioaerosols from the wastewater treatment plant can be propagated into the air, affecting the health of residents, animals, and plants around the area. This study was conducted to evaluate the density and composition of bioaerosols (bacteria and fungi) in anoxic and aeration tanks of a domestic wastewater treatment plant. Bioaerosol samples were taken by the active method with SKC Biostage. The agar dishes were placed at a height of 1.5 m above the ground. After sample collection, the dishes were incubated at 37 ±1 °C for 24h-48h for bacteria and at 25 ±1 °C for 72h-120h for fungi. Then, these microorganisms were identified by 16S rRNA (16S Metabarcoding) method for bacteria and taxonomy method for fungi. The results showed that the density of bacteria and mold in the anoxic tank ranged from 233.2 ±7.8 to 339.2 ±13.9 CFU/m3 and 91.8 ±3.2 to 197.9 ±10.1 CFU/m3, respectively. Meanwhile, bacterial and mold densities of aeration tank ranged from 275.6 ±10.4 to 424.0 ±17.3 CFU/m3 and 176.7 ±5.8 to 254.4 ±12.1 CFU/m3, respectively. The density of bacteria and mold in the aeration tank is higher than in the anoxic tank. On the other hand, the 16S Metabarcoding analysis for the microbial diversity detects more than 100 species in two tanks, the predominant bacterial species are: Bacillus megaterium (B. megaterium), Bacillus cereus (B. cereus), Staphylococcus sciuri (S. sciuri) and two common fungi Aspergillus tamarii (A. tamarii), and Penicillium rubers (P. rubers). This preliminary survey in Viet Nam reveals the existence and diversity of bioaerosols in the air from the biological treatment tanks of the wastewater treatment plant.

Monitoring of enteropathogenic Gram-negative bacteria in wastewater treatment plants: a multimethod approach

The wastewater treatment processes are associated with the emission of microbial aerosols, including enteropathogenic bacteria. Their presence in this work environment poses a real threat to the health of employees, both through the possibility of direct inhalation of the contaminated air and indirectly through the pollution of all types of surfaces with such bioaerosol particles. This study aimed to investigate the prevalence of enteropathogenic bacteria in the air, on surfaces, and in wastewater samples collected in four wastewater treatment plants (WWTPs). The effectiveness of conventional culture-biochemical, as well as spectrometric and molecular methods for the rapid detection of enteropathogenic bacteria at workstations related to particular stages of wastewater processing, was also evaluated. Bioaerosol, surface swab, and influent and effluent samples were collected from wastewater plants employing mechanical–biological treatment technologies. The air samples were collected using MAS-100 NT impactor placed at a height of 1.5 m above the floor or ground, simulating aspiration from the human breathing zone. Surface samples were collected with sterile swabs from different surfaces (valves, handles, handrails, and coveyor belts) at workplaces. The raw influent and treated effluent wastewater samples were aseptically collected using sterile bottles. The identification of bacterial entheropathogens was simultaneously conducted using a culture-based method supplemented with biochemical (API) tests, mass-spectrometry (MALDI TOF MS), and molecular (multiplex real-time PCR) methods. This study confirmed the common presence of bacterial pathogens (including enteropathogenic and enterotoxigenic Escherichia coli, Salmonella spp., Campylobacter spp., and Yersinia enterocolitica) in all air, surface, and wastewater samples at studied workplaces. Higher concentrations of enteropathogenic bacteria were observed in the air and on surfaces at workplaces where treatment processes were not hermetized. The results of this study underline that identification of enteropathogenic bacteria in WWTPs is of great importance for the correct risk assessment at workplaces. From the analytical point of view, the control of enteropathogenic bacterial air and surface pollution using rapid multiplex-PCR method should be routinely performed as a part of hygienic quality assessment in WWTPs.

Characteristics and health impacts of bioaerosols in animal barns: A comprehensive study

Bioaerosols produced during animal production have potential adverse effects on the health of workers and animals. Our objective was to investigate characteristics, antibiotic-resistance genes (ARGs), and health risks of bioaerosols in various animal barns. Poultry and swine barns had high concentrations of airborne bacteria (11156 and 10917 CFU/m3, respectively). Acinetobacter, Clostridium sensu stricto, Corynebacterium, Pseudomonas, Psychrobacter, Streptococcus, and Staphylococcus were dominant pathogenic bacteria in animal barns, with Firmicutes being the most abundant bacterial phylum. Based on linear discriminant analysis effect size (LEfSe), there were more discriminative biomarkers in cattle barns than in poultry or swine barns, although the latter had the highest abundance of bacterial pathogens and high abundances of ARGs (including tetM, tetO, tetQ, tetW sul1, sul2, ermA, ermB) and intI1). Based on network analyses, there were higher co-occurrence patterns between bacteria and ARGs in bioaerosol from swine barns. Furthermore, in these barns, relative abundance of bacteria in bioaerosol samples was greatly affected by environmental factors, mainly temperature, relative humidity, and concentrations of CO2, NH3, and PM2.5. This study provided novel data regarding airborne bio-contaminants in animal enclosures and an impetus to improve management to reduce potential health impacts on humans and animals.

Eco-Friendly Sanitization of Indoor Environments: Effectiveness of Thyme Essential Oil in Controlling Bioaerosol Levels and Disinfecting Surfaces.

Bioaerosols and pathogens in indoor workplaces and residential environments are the primary culprits of several infections. Techniques for sanitizing air and surfaces typically involve the use of UV rays or chemical sanitizers, which may release chemical residues harmful to human health. Essential oils, natural substances derived from plants, which exhibit broad antimicrobial properties, could be a viable alternative for air and surface sanitation. The objective of this study has been to investigate the efficacy of thyme essential oil (TEO) in environmental sanitation processes. In Vitro assays through agar well diffusion, disk volatilization and tube dilution methods revealed significant antimicrobial activity of TEO 100% against foodborne and environmental isolates, with both bacteriostatic/fungistatic and bactericidal/fungicidal effects. Therefore, aqueous solutions of TEO 2.5% and 5% were formulated for air sanitation through nebulization and surface disinfection via direct contact. Bioaerosol samples and surface swabs were analyzed before and after sanitation, demonstrating the efficacy of aqueous solutions of TEO in reducing mesophilic and psychrophilic bacteria and environmental fungi levels in both air and on surfaces. The obtained results prove the antimicrobial potential of aqueous solutions of TEO in improving indoor air quality and surface cleanliness, suggesting thyme essential oil as an effective and safe natural sanitizer with minimal environmental impact compared to dangerous chemical disinfectants.