Versatile filter membrane for effective sampling and real-time quantitative detection of airborne pathogens.

Construction of air filter membranes bearing prominent collecting and transferring capability is highly desirable for detecting airborne pathogens but remains challenging. Here, a hyaluronic acid air filter membrane (HAFM) with tunable heterogeneous micro-nano porous structures is straightforwardly constructed through the ethanol-induced phase separation strategy. Airborne pathogens can be trapped and collected by HAFM with high performance due to the ideal trade-off between removal efficiency and pressure drop. By exempting the sample elution and extraction processes, the HAFM after filtration sampling can not only directly disperse on the agar plate for colony culture but also turn to an aqueous solution for centrifugal enrichment, which significantly reduces the damage and losses of the captured microorganisms. The following combination with ATP bioluminescence endows the HAFM with a real-time quantitative detection function for the captured airborne pathogens. Benefiting from high-efficiency sampling and non-traumatic transfer of airborne pathogens, the real-world bioaerosol concentration can be facilely evaluated by the HAFM-based ATP assay. This work thus not only provides a feasible strategy to fabricate air filter membranes for efficient microbial collection and enrichment but also sheds light on designing advanced protocols for real-time detection of bioaerosols in the field.

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.

Comparison of Plenum and Cage-level Filter Exhaust Dust PCR Testing to Soiled Bedding Sentinel Mice (Mus musculus) on an IVC Rack.

The use of soiled-bedded sentinels (SBSs) has historically been the standard for colony health surveillance monitoring at our institution. With the advent of newer technologies in which dust collected from filters is tested by PCR, we compared traditional SBS with PCR testing of both exhaust air dust collected from a filter in the downstream vertical plenum (exhaust dust test [EDT]) and the SBS cage-level exhaust filter (SCEF). Our hypothesis was that both methods of filter testing would identify more pathogens than SBS testing. Twenty-five individually ventilated mouse racks that used disposable caging were sanitized and placed into rotation. Rack plenums were tested by PCR to verify negative results before the study start. Exhaust dust collection media were placed in the exhaust plenum (n = 25). SBS cages were placed on each side of the rack with 2 mice per cage (n = 42 mice), with the remaining cage slots occupied by research animals. At each triweekly cage change, the exhaust air filters were carefully removed from the cage top, placed in sterile 50-mL conical tubes, and pooled for submission. After 3mo, the SBS mice were tested via serology for bacterial and viral agents and by PCR for Helicobacter species, pinworms, and ectoparasites. In addition, the EDT filter and SCEF were collected for PCR to evaluate for the same agents. Our results indicate that the SCEF consistently detected agents more frequently than the EDT filter placed in the plenum and that the EDT filter media detected agents more frequently than did the SBS mice. Our data suggest that both PCR methods of detection are superior to SBS for individually ventilated disposable rodent cages and that the SCEF is superior to EDT. These data supported our movement of institution toward environmental monitoring as a method of rodent colony health surveillance.

Relationship Between Airborne Fungi Presence and the Position of the High Efficiency Particulate Air Filter in the Heating, Ventilation, and Air Conditioning System.

AIM: Establish the influence of the terminal or nonterminal position of High Efficiency Particulate Air (HEPA) filters in the Heating, Ventilation, and Air Conditioning (HVAC) system on the presence of airborne fungi in controlled environment rooms. BACKGROUND: Fungal infections are an important cause of morbidity and mortality in hospitalized patients. METHODS: This study was realized from 2010 to 2017, in rooms with terminal and nonterminal HEPA filters, in eight Spanish hospitals. In rooms with terminal HEPA filters, 2,053 and 2,049 samples were recollected, and in rooms with nonterminal HEPA filters, 430 and 428 samples were recollected in the air discharge outlet (Point 1) and in the center of the room (Point 2), respectively. Temperature, relative humidity, air changes per hour, and differential pressure were recollected. RESULTS: Multivariable analysis showed higher odds ratio (OR) of airborne fungi presence when HEPA filters were in nonterminal position (OR: 6.78; 95% CI [3.77, 12.20]) in Point 1 and (OR: 4.43; 95% CI [2.65, 7.40]) in Point 2. Other parameters influenced airborne fungi presence, such as temperature (OR: 1.23; 95% CI [1.06, 1.41]) in Point 2 differential pressure (OR: 0.86; 95% CI [0.84, 0.90]) and (OR: 0.88; 95% CI [0.86, 0.91]) in Points 1 and 2, respectively. CONCLUSIONS: HEPA filter in terminal position of the HVAC system reduces the presence of airborne fungi. To decrease the presence of airborne fungi, adequate maintenance of the environmental and design parameters is necessary in addition to the terminal position of the HEPA filter.

Efficacy of antimicrobial and anti-viral coated air filters to prevent the spread of airborne pathogens.

The COVID-19 pandemic has demonstrated the real need for mechanisms to control the spread of airborne respiratory pathogens. Thus, preventing the spread of disease from pathogens has come to the forefront of the public consciousness. This has brought an increasing demand for novel technologies to prioritise clean air. In this study we report on the efficacy of novel biocide treated filters and their antimicrobial activity against bacteria, fungi and viruses. The antimicrobial filters reported here are shown to kill pathogens, such as Candida albicans, Escherichia coli and MRSA in under 15 min and to destroy SARS-CoV-2 viral particles in under 30 s following contact with the filter. Through air flow rate testing, light microscopy and SEM, the filters are shown to maintain their structure and filtration function. Further to this, the filters are shown to be extremely durable and to maintain antimicrobial activity throughout the operational lifetime of the product. Lastly, the filters have been tested in field trials onboard the UK rail network, showing excellent efficacy in reducing the burden of microbial species colonising the air conditioning system.

Positive and Negative Ions Potently Inhibit the Viability of Airborne Gram-Positive and Gram-Negative Bacteria.

Positive and negative ions (PAIs and NAIs, respectively) generated by air ionizers curb indoor spread of airborne pathogens through cellular oxidative damage. Thus, here, we asked whether ion exposure of Staphylococcus aureus and Escherichia coli bacteria-either plated on agar or trapped in air filters-would affect their viability and whether this effect would be influenced by variations in bacterial type and load, action area, distance from the ion generator, exposure time, or filter type. We selected these two vegetative bacterium species because, besides being representative of Gram-positive and Gram-negative strains, respectively, they are widely recognized as the two most common airborne pathogens. We observed a robust ion inhibitory effect on the viability of free bacteria regardless of the experimental condition employed. Specifically, 12-h ion exposure of plated S. aureus and E. coli, at either 5 cm or 10 cm from the ion source, reduced bacterial viability by ∼95% and 70%, respectively. Furthermore, 3-h ion exposure was sufficient to reduce the viability of both bacterial species trapped in filters. Our results showing a strong antibacterial activity of PAI and NAI under all experimental conditions tested further support the use of air ionizers for preventing and/or containing airborne infection in domestic and nondomestic settings. IMPORTANCE Indoor air is a well-established vehicle for direct and indirect spread of a wide variety of human pathogens-as bioaerosols are composed of bacteria, viruses, fungi, and other types of organisms-that may trigger some pathologies. Plasmacluster ionizers are known for their ability to generate positively or negatively charged air ions (PAIs and NAIs, respectively) that can kill/inactivate indoor airborne pathogens, through oxidative stress-induced damage, in various environments. Given these premises, the aim of this study was to evaluate the viability of Gram-positive and Gram-negative bacteria exposed to PAI and NAI under different experimental variables such as bacterial type and load, action area, distance from the ion generator, ion exposure time, and filter type. Altogether, our findings, demonstrating a remarkable PAI and NAI antibacterial activity, stress the importance of using air ionizers to prevent indoor airborne infection.

Novel non intrusive continuous use ZeBox technology to trap and kill airborne microbes.

Preventing nosocomial infection is a major unmet need of our times. Existing air decontamination technologies suffer from demerits such as toxicity of exposure, species specificity, noxious gas emission, environment-dependent performance and high power consumption. Here, we present a novel technology called "ZeBox" that transcends the conventional limitations and achieves high microbicidal efficiency. In ZeBox, a non-ionizing electric field extracts naturally charged microbes from flowing air and deposits them on engineered microbicidal surfaces. The surface's three dimensional topography traps the microbes long enough for them to be inactivated. The electric field and chemical surfaces synergistically achieve rapid inactivation of a broad spectrum of microbes. ZeBox achieved near complete kill of airborne microbes in challenge tests (5-9 log reduction) and [Formula: see text] efficiency in a fully functional stem cell research facility in the presence of humans. Thus, ZeBox fulfills the dire need for a real-time, continuous, safe, trap-and-kill air decontamination technology.

Triboelectrification induced self-powered microbial disinfection using nanowire-enhanced localized electric field.

Air-transmitted pathogens may cause severe epidemics showing huge threats to public health. Microbial inactivation in the air is essential, whereas the feasibility of existing air disinfection technologies meets challenges including only achieving physical separation but no inactivation, obvious pressure drops, and energy intensiveness. Here we report a rapid disinfection method toward air-transmitted bacteria and viruses using the nanowire-enhanced localized electric field to damage the outer structures of microbes. This air disinfection system is driven by a triboelectric nanogenerator that converts mechanical vibration to electricity effectively and achieves self-powered. Assisted by a rational design for the accelerated charging and trapping of microbes, this air disinfection system promotes microbial transport and achieves high performance: >99.99% microbial inactivation within 0.025 s in a fast airflow (2 m/s) while only causing low pressure drops (<24 Pa). This rapid, self-powered air disinfection method may fill the urgent need for air-transmitted microbial inactivation to protect public health.

Should homes and workplaces purchase portable air filters to reduce the transmission of SARS-CoV-2 and other respiratory infections? A systematic review.

Respiratory infections, including SARS-CoV-2, are spread via inhalation or ingestion of airborne pathogens. Airborne transmission is difficult to control, particularly indoors. Manufacturers of high efficiency particulate air (HEPA) filters claim they remove almost all small particles including airborne bacteria and viruses. This study investigates whether modern portable, commercially available air filters reduce the incidence of respiratory infections and/or remove bacteria and viruses from indoor air. We systematically searched Medline, Embase and Cochrane for studies published between January 2000 and September 2020. Studies were eligible for inclusion if they included a portable, commercially available air filter in any indoor setting including care homes, schools or healthcare settings, investigating either associations with incidence of respiratory infections or removal and/or capture of aerosolised bacteria and viruses from the air within the filters. Dual data screening and extraction with narrative synthesis. No studies were found investigating the effects of air filters on the incidence of respiratory infections. Two studies investigated bacterial capture within filters and bacterial load in indoor air. One reported higher numbers of viable bacteria in the HEPA filter than in floor dust samples. The other reported HEPA filtration combined with ultraviolet light reduced bacterial load in the air by 41% (sampling time not reported). Neither paper investigated effects on viruses. There is an important absence of evidence regarding the effectiveness of a potentially cost-efficient intervention for indoor transmission of respiratory infections, including SARS-CoV-2. Two studies provide 'proof of principle' that air filters can capture airborne bacteria in an indoor setting. Randomised controlled trials are urgently needed to investigate effects of portable HEPA filters on incidence of respiratory infections.

Construction and validation of UV-C decontamination cabinets for filtering facepiece respirators.

We present evidence-based design principles for three different UV-C based decontamination systems for N95 filtering facepiece respirators (FFRs) within the context of the SARS-CoV-2 outbreak of 2019-2020. The approaches used here were created with consideration for the needs of low- and middle-income countries (LMICs) and other under-resourced facilities. As such, a particular emphasis is placed on providing cost-effective solutions that can be implemented in short order using generally available components and subsystems. We discuss three optical designs for decontamination chambers, describe experiments verifying design parameters, validate the efficacy of the decontamination for two commonly used N95 FFRs (3M, #1860 and Gerson #1730), and run mechanical and filtration tests that support FFR reuse for at least five decontamination cycles.

Salt coatings functionalize inert membranes into high-performing filters against infectious respiratory diseases.

Respiratory protection is key in infection prevention of airborne diseases, as highlighted by the COVID-19 pandemic for instance. Conventional technologies have several drawbacks (i.e., cross-infection risk, filtration efficiency improvements limited by difficulty in breathing, and no safe reusability), which have yet to be addressed in a single device. Here, we report the development of a filter overcoming the major technical challenges of respiratory protective devices. Large-pore membranes, offering high breathability but low bacteria capture, were functionalized to have a uniform salt layer on the fibers. The salt-functionalized membranes achieved high filtration efficiency as opposed to the bare membrane, with differences of up to 48%, while maintaining high breathability (> 60% increase compared to commercial surgical masks even for the thickest salt filters tested). The salt-functionalized filters quickly killed Gram-positive and Gram-negative bacteria aerosols in vitro, with CFU reductions observed as early as within 5 min, and in vivo by causing structural damage due to salt recrystallization. The salt coatings retained the pathogen inactivation capability at harsh environmental conditions (37 °C and a relative humidity of 70%, 80% and 90%). Combination of these properties in one filter will lead to the production of an effective device, comprehensibly mitigating infection transmission globally.

Bacterial community analysis of floor dust and HEPA filters in air purifiers used in office rooms in ILAS, Beijing.

Air purifiers with high-efficiency particulate air (HEPA) filters remove not only particulate matter but also airborne microorganisms in indoor environments. We investigated the bacterial community in HEPA filters (used for 1 year) and that in the floor dust of 12 office rooms in Beijing. We found that the viable bacteria proportion in the filter was significantly higher than that in the floor dust (p < 0.001). The Non-Metric Multi-Dimensional Scaling analysis showed that the bacterial communities in the filters and dust were significantly different (p = 0.001). The Chao1, Shannon-Wiener and phylogenetic diversity values in the filter were significantly higher than those in the dust (p < 0.001). The predominant bacterial classes in the filter were Alphaproteobacteria and Actinobacteria, whereas those in the dust were Bacteroidia, Clostridia and Bacilli. Human occupancy contributed more to the bacterial community in the filter than that in the dust. Klebsiella and Alloprevotella in the dust and filters positively correlated with the occupancy density. Soil bacteria contributed to a significantly higher proportion of the bacteria in the HEPA filter (p < 0.001). In contrast, human oral, indoor air and outdoor haze contributed to a higher proportion of the bacteria in the dust samples (p < 0.001, p < 0.01 and p < 0.05, respectively). As HEPA filters serve as an ecological niche for indoor bacteria, they should be carefully investigated during the assessment of indoor environmental health.

Enhancement of using combined packing materials on the removal of mixed sulfur compounds in a biotrickling filter and analysis of microbial communities.

BACKGROUND: Packing materials is a critical design consideration when employing biological reactor to treat malodorous gases. The acidification of packing bed usually results in a significant drop in the removal efficiency. In the present study, a biotrickling filter (BTF2) packed with plastic balls in the upper layer and with lava rocks in the bottom layer, was proposed to mitigate the acidification. RESULTS: Results showed that using combined packing materials efficiently enhanced the removal performance of BTF2 when compared with BTF1, which was packed with sole lava rocks. Removal efficiencies of more than 92.5% on four sulfur compounds were achieved in BTF2. Average pH value in its bottom packing bed was about 4.86, significantly higher than that in BTF1 (2.85). Sulfate and elemental sulfur were observed to accumulate more in BTF1 than in BTF2. Analysis of principal coordinate analysis proved that structure of microbial communities in BTF2 changed less after the shutdown but more when the initial pH value was set at 5.5. Network analysis of significant co-occurrence patterns based on the correlations between microbial taxa revealed that BTF2 harbored more diverse microorganisms involving in the bio-oxidation of sulfur compounds and had more complex interactions between microbial species. CONCLUSIONS: Results confirmed that using combined packing materials effectively improved conditions for the growth of microorganisms. The robustness of reactor against acidification, adverse temperature and gas supply shutdown was greatly enhanced. These provided a theoretical basis for using mixed packing materials to improve removal performance.

Long-term microbial community dynamics at two full-scale biotrickling filters treating pig house exhaust air.

In this study, the microbial community structure of two full-scale biotrickling filters treating exhaust air from a pig housing facility were evaluated using 16S metabarcoding. The effect of inoculation with activated sludge of a nearby domestic waste water treatment plant was investigated, which is a cheap procedure and easy to apply in practice. The study was performed at a three-stage and a two-stage full-scale biotrickling filter; of which, only the latter was inoculated. Both biotrickling filters evolved towards a rather similar community over time, which differed from the one in the activated sludge used for inoculation. However, the bacterial population at both biotrickling filters showed small differences on the family level. A large population of heterotrophic bacteria, including denitrifying bacteria, was present in both biotrickling filters. In the non-inoculated biotrickling filter, nitrite-oxidizing bacteria (NOB) could not be detected, which corresponded with the incomplete nitrification leading to high nitrite accumulation observed in this system. Inoculation with the wide spectrum inoculum activated sludge had in this study a positive effect on the biotrickling filter performance (higher ammonia removal and lower nitrous oxide production). It could thus be beneficial to inoculate biotrickling filters in order to enrich NOB at the start-up, making it easier to keep the free nitrous acid concentration low enough to not be inhibited by it.

Inactivation of bacterial and fungal spores by UV irradiation and gaseous iodine treatment applied to air handling filters.

Exposure to viable bacterial and fungal spores re-aerosolized from air handling filters may create a major health risk. Assessing and controlling this exposure have been of interest to the bio-defense and indoor air quality communities. Methods are being developed for inactivating stress-resistant viable microorganisms collected on ventilation filters. Here we investigated the inactivation of spores of Bacillus thuringiensis var. kurstaki (Btk), a recognized simulant for B. antracis, and Aspergillus fumigatus, a common opportunistic pathogen used as an indicator for indoor air quality. The viability change was measured on filters treated with ultraviolet (UV) irradiation and gaseous iodine. The spores were collected on high-efficiency particulate air (HEPA) and non-HEPA filters, both flattened for testing purposes to represent "surface" filters. A mixed cellulose ester (MCE) membrane filter was also tested as a reference. Additionally, a commercial HEPA unit with a deep-bed (non-flattened) filter was tested. Combined treatments of Btk spores with UV and iodine on MCE filter produced a synergistic inactivation effect. No similar synergy was observed for A. fumigatus. For spores collected on an MCE filter, the inactivation effect was about an order of magnitude greater for Btk compared to A. fumigatus. The filter type was found to be an important factor affecting the inactivation of Btk spores while it was not as influential for A. fumigatus. Overall, the combined effect of UV irradiation and gaseous iodine on viable bacterial and fungal spores collected on flat filters was found to be potent. The benefit of either simultaneous or sequential treatment was much lower for Btk spores embedded inside the deep-bed (non-flattened) HEPA filter, but for A. fumigatus the inactivation on flattened and non-flattened HEPA filters was comparable. For both species, applying UV first and gaseous iodine second produced significantly higher inactivation than when applying them simultaneously or in an opposite sequence.

Prevalence of Antibiotic Resistance Genes in Air-Conditioning Systems in Hospitals, Farms, and Residences.

High-throughput quantitative PCR combined with Illumina sequencing and network analysis were used to characterize the antibiotic resistance gene (ARG) profiles in air-conditioning filters from different environments. In total, 177 ARGs comprising 10 ARG types were determined. The detectable numbers and the relative abundance of ARGs in hospitals and farms were significantly higher than those in city and village residences. Compared to hospitals, farms had a higher level of tetracycline, multidrug, integrase, and macrolide⁻lincosamide⁻streptogramin (MLS) B resistance genes but a lower level of beta-lactam resistance genes. The bl3_cpha gene was the most abundant resistance gene subtype in hospital samples with an abundance of 2.01 × 10-4 copies/16S rRNA, while a level of only 5.08 × 10-12 copies/16S rRNA was observed in farm samples. There was no significant difference in bacterial diversity among the hospitals, farms, and residences, and Proteobacteria was the most abundant phylum. Network analysis revealed that Proteobacteria and Actinobacteria were possible hosts of the beta-lactam, MLSB, aminoglycoside, multidrug, sulfonamide, and tetracycline resistance genes. The results demonstrate that ARGs exist in indoor environments and that farms and hospitals are important sources. This study provides a useful reference for understanding the distribution patterns and risk management of ARGs in indoor environments.

Quantitative filter forensics with residential HVAC filters to assess indoor concentrations.

Analysis of the dust from heating, ventilation, and air conditioning (HVAC) filters is a promising long-term sampling method to characterize airborne particle-bound contaminants. This filter forensics (FF) approach provides valuable insights about differences between buildings, but does not allow for an estimation of indoor concentrations. In this investigation, FF is extended to quantitative filter forensics (QFF) by using measurements of the volume of air that passes through the filter and the filter efficiency, to assess the integrated average airborne concentrations of total fungal and bacterial DNA, 36 fungal species, endotoxins, phthalates, and organophosphate esters (OPEs) based on dust extracted from HVAC filters. Filters were collected from 59 homes located in central Texas, USA, after 1 month of deployment in each summer and winter. Results showed considerable differences in the concentrations of airborne particle-bound contaminants in studied homes. The airborne concentrations for most of the analytes are comparable with those reported in the literature. In this sample of homes, the HVAC characterization measurements varied much less between homes than the variation in the filter dust concentration of each analyte, suggesting that even in the absence of HVAC data, FF can provide insight about concentration differences for homes with similar HVAC systems.

Effectiveness of Airborne Fungi Removal by using a HEPA Air Purifier Fan in Houses.

Few studies have evaluated the performance of air purifiers in removing airborne fungi in houses. Here, we evaluated the ability of a HEPA air purifier fan to remove airborne fungi in six houses in Japan. In each house, the number of airborne fungi decreased more rapidly when the air purifier fan was on (test measurement) than when it was off (control) , demonstrating its ability to decrease the fungal concentration. The number of airborne fungi decreased between 1.5 and 6 times faster when the air purifier fan was on than when it was off (spontaneous decrease) . Clean air change rates, calculated from measurements taken 15 min after the test equipment operation began, ranged from 2.9 to 5.4 (h-1) , indicating adequate air cleaning. One of the six test houses contained a much greater concentration of airborne fungi than the standard set by the Architectural Institute of Japan. When the air purifier fan was operated in the house, the indoor/outdoor (I/O) ratio decreased from 77.5, equating to a fungal concentration of 53,000 cfu/m3 at 0 min to 0.72 or 620 cfu/m3 after 45 min, which is below the standard. This reduction clearly demonstrated the antifungal effect of the air purifier fan.