Real-Time Monitoring of Air Pollution Health Impacts Using Breath-Borne Gaseous Biomarkers from Rats.

Offline techniques are adopted for studying air pollution health impacts, thus failing to provide in situ observations. Here, we have demonstrated their real-time monitoring by online analyzing an array of gaseous biomarkers from rats' exhaled breath using an integrated exhaled breath array sensor (IEBAS) developed. The biomarkers include total volatile organic compounds (TVOC), CO2, CO, NO, H2S, H2O2, O2, and NH3. Specific breath-borne VOCs were also analyzed by a gas chromatography-ion mobility spectrometer (GC-IMS). After real-life ambient air pollution exposures (2 h), the pollution levels of PM2.5 and O3 were both found to significantly affect the relative levels of multiple gaseous biomarkers in rats' breath. Eleven biomarkers, especially NO, H2S, and 1-propanol, were detected as significantly correlated with PM2.5 concentration, while heptanal was shown to be significantly correlated with O3. Likewise, significant changes were also detected in multiple breath-borne biomarkers from rats under lab-controlled O3 exposures with levels of 150, 300, and 1000 µg/m3 (2 h), compared to synthetic air exposure. Importantly, heptanal was experimentally confirmed as a reliable biomarker for O3 exposure, with a notable dose-response relationship. In contrast, conventional biomarkers of inflammation and oxidative stress in rat sera exhibited insignificant differences after the 2 h exposures. The results imply that breath-borne gaseous biomarkers can serve as an early and sensitive indicator for ambient pollutant exposure. This work pioneered a new research paradigm for online monitoring of air pollution health impacts while obtaining important candidate biomarker information for PM2.5 and O3 exposures.

A Near-Explicit Reaction Mechanism of Chlorine-Initiated Limonene: Implications for Health Risks Associated with the Concurrent Use of Cleaning Agents and Disinfectants.

Limonene, a key volatile chemical product (VCP) commonly found in personal care and cleaning agents, is emerging as a major indoor air pollutant. Recently, elevated levels of reactive chlorine species during bleach cleaning and disinfection have been reported to increase indoor oxidative capacity. However, incomplete knowledge of the indoor transformation of limonene, especially the missing chlorine chemistry, poses a barrier to evaluating the environmental implications associated with the concurrent use of cleaning agents and disinfectants. Here, we investigated the reaction mechanisms of chlorinated limonene peroxy radicals (Cl-lim-RO2•), key intermediates in determining the chlorine chemistry of limonene, and toxicity of transformation products (TPs) using quantum chemical calculations and toxicology modeling. The results indicate that Cl-lim-RO2• undergoes a concerted autoxidation process modulated by RO2• and alkoxy radicals (RO•), particularly emphasizing the importance of RO• isomerization. Following this generalized autoxidation mechanism, Cl-lim-RO2• can produce low-volatility precursors of secondary organic aerosols. Toxicological findings further indicate that the majority of TPs exhibit increased respiratory toxicity, mutagenicity, and eye/skin irritation compared to limonene, presenting an occupational hazard for indoor occupants. The proposed near-explicit reaction mechanism of chlorine-initiated limonene significantly enhances our current understanding of both RO2• and RO• chemistry while also highlighting the health risks associated with the concurrent use of cleaning agents and disinfectants.

On-site airborne pathogen detection for infection risk mitigation.

Human-infecting pathogens that transmit through the air pose a significant threat to public health. As a prominent instance, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that caused the COVID-19 pandemic has affected the world in an unprecedented manner over the past few years. Despite the dissipating pandemic gloom, the lessons we have learned in dealing with pathogen-laden aerosols should be thoroughly reviewed because the airborne transmission risk may have been grossly underestimated. From a bioanalytical chemistry perspective, on-site airborne pathogen detection can be an effective non-pharmaceutic intervention (NPI) strategy, with on-site airborne pathogen detection and early-stage infection risk evaluation reducing the spread of disease and enabling life-saving decisions to be made. In light of this, we summarize the recent advances in highly efficient pathogen-laden aerosol sampling approaches, bioanalytical sensing technologies, and the prospects for airborne pathogen exposure measurement and evidence-based transmission interventions. We also discuss open challenges facing general bioaerosols detection, such as handling complex aerosol samples, improving sensitivity for airborne pathogen quantification, and establishing a risk assessment system with high spatiotemporal resolution for mitigating airborne transmission risks. This review provides a multidisciplinary outlook for future opportunities to improve the on-site airborne pathogen detection techniques, thereby enhancing the preparedness for more on-site bioaerosols measurement scenarios, such as monitoring high-risk pathogens on airplanes, weaponized pathogen aerosols, influenza variants at the workplace, and pollutant correlated with sick building syndromes.

Quantitatively Visualizing Airborne Disease Transmission Risks of Different Exhalation Activities through CO2 Imaging.

Aerosol transmission has played a leading role in COVID-19 pandemic. However, there is still a poor understanding about how it is transmitted. This work was designed to study the exhaled breath flow dynamics and transmission risks under different exhaling modes. Using an infrared photography device, exhaled flow characteristics of different breathing activities, such as deep breathing, dry coughing, and laughing, together with the roles of mouth and nose were characterized by imaging CO2 flow morphologies. Both mouth and nose played an important role in the disease transmission though in the downward direction for the nose. In contrast to the trajectory commonly modeled, the exhaled airflows appeared with turbulent entrainments and obvious irregular movements, particularly the exhalations involving mouth were directed horizontal and had a higher propagation capacity and transmission risk. While the cumulative risk was high for deep breathing, those transient ones from dry coughing, yawning, and laughing were also shown to be significant. Various protective measures including masks, canteen table shields, and wearable devices were visually demonstrated to be effective for altering the exhaled flow directions. This work is useful to understanding the risk of aerosol infection and guiding the formulation of its prevention and control strategies. Experimental data also provide important information for refining model boundary conditions.

Exhaled VOC Biomarkers from Rats Injected with PMs from Thirty-One Major Cities in China.

Particulate matter (PMs) of different origins can cause diverse health effects. Here, a homemade box was used to facilitate real-time measurements of breath-borne volatile organic compounds (VOCs) by gas chromatography-ion mobility spectrometry. We have tracked exhaled VOC changes in 228 Wistar rats that were injected with water-soluble PM suspension filtrates (after 0.45 µm) from 31 China cities for 1 h to up to 1-6 days during the experiments. Rats exposed to the filtrates exhibited significant changes in breath-borne VOCs within hours, featuring dynamic fluctuations in the levels of acetone, butan-2-one, heptan-2-one-M, acetic acid-M, and ethanol. Subsequently, on the fifth to sixth day after the injection, there was a notable increase in the proportion of aldehydes (including hexanal-M, hexanal-D, pentanal, heptanal-M, and (E)-2-hexenal). The 10 dynamic VOC fingerprint patterns mentioned earlier showcased the capability to indirectly differentiate urban PM toxicity and categorize the 31 cities into four distinct groups based on their health effects. This study provides valuable insights into the mechanisms of exhaled VOCs and underscores their critical role as biomarkers for differentiating the toxicity of different PMs and detecting the early signs of potential diseases. The results from this work also provide a scientific basis for city-specific air pollution control and policy development.

Haze Air Pollution Health Impacts of Breath-Borne VOCs.

Here, we investigated the use of breath-borne volatile organic compounds (VOCs) for rapid monitoring of air pollution health effects on humans. Forty-seven healthy college students were recruited, and their exhaled breath samples (n = 235) were collected and analyzed for VOCs before, on, and after two separate haze pollution episodes using gas chromatography-ion mobility spectrometry (GC-IMS). Using a paired t-test and machine learning model (Gradient Boosting Machine, GBM), six exhaled VOC species including propanol and isoprene were revealed to differ significantly among pre-, on-, and post-exposure in both haze episodes, while none was found between clean control days. The GBM model was shown capable of differentiating between pre- and on-exposure to haze pollution with a precision of 90-100% for both haze episodes. However, poor performance was detected for the same model between two different clean days. In addition to gender and particular haze occurrence influences, correlation analysis revealed that NH4+, NO3-, acetic acid, mesylate, CO, NO2, PM2.5, and O3 played important roles in the changes in breath-borne VOC fingerprints following haze air pollution exposure. This work has demonstrated direct evidence of human health impacts of haze pollution while identifying potential breath-borne VOC biomarkers such as propanol and isoprene for haze air pollution exposure.

Gene-Regulated Release of Distinctive Volatile Organic Compounds from Stressed Living Cells.

Breath-borne volatile organic compounds (VOCs) have been increasingly studied as non-invasive biomarkers in both medical diagnosis and environmental health research. Recently, changes in breath-borne VOC fingerprints were demonstrated in rats and humans following pollutant exposures. In this study, the eukaryotic model Saccharomyces cerevisiae was used to study the release of cellular VOCs resulting from toxicant exposures (i.e., O3, H2O2, and CO2) and its underlying biological mechanism. Our results showed that different toxicant exposures caused the release of distinctive VOC profiles of yeast cells. The levels of ethyl acetate and ethyl n-propionate were altered in response to all the toxicants used in this study and could thus be targeted for future environmental toxicity monitoring. The RNA-seq results revealed significant changes in the metabolic or signaling pathways related to the ribosome, carbohydrate, and amino acid metabolisms after exposures. Notably, the shift from glycolysis to the pentose phosphate pathway of carbohydrate metabolism and the inhabitation of the aspartate pathway in the lysine synthesis was essential to the cellular antioxidation by providing reduced nicotinamide adenine dinucleotide phosphate (NADPH). The reprogrammed metabolisms could have resulted in the observed changes of VOCs released, e.g., the production of ethyl acetate for detoxification from yeast cells. This study provides further evidence that VOCs released from living organisms could be used to monitor and guard against toxic exposures while providing better mechanistic insights of the changes in breath-borne VOCs previously observed in rats and humans exposed to air toxicants.

What were the historical reasons for the resistance to recognizing airborne transmission during the COVID-19 pandemic?

The question of whether SARS-CoV-2 is mainly transmitted by droplets or aerosols has been highly controversial. We sought to explain this controversy through a historical analysis of transmission research in other diseases. For most of human history, the dominant paradigm was that many diseases were carried by the air, often over long distances and in a phantasmagorical way. This miasmatic paradigm was challenged in the mid to late 19th century with the rise of germ theory, and as diseases such as cholera, puerperal fever, and malaria were found to actually transmit in other ways. Motivated by his views on the importance of contact/droplet infection, and the resistance he encountered from the remaining influence of miasma theory, prominent public health official Charles Chapin in 1910 helped initiate a successful paradigm shift, deeming airborne transmission most unlikely. This new paradigm became dominant. However, the lack of understanding of aerosols led to systematic errors in the interpretation of research evidence on transmission pathways. For the next five decades, airborne transmission was considered of negligible or minor importance for all major respiratory diseases, until a demonstration of airborne transmission of tuberculosis (which had been mistakenly thought to be transmitted by droplets) in 1962. The contact/droplet paradigm remained dominant, and only a few diseases were widely accepted as airborne before COVID-19: those that were clearly transmitted to people not in the same room. The acceleration of interdisciplinary research inspired by the COVID-19 pandemic has shown that airborne transmission is a major mode of transmission for this disease, and is likely to be significant for many respiratory infectious diseases.

Coronavirus Disease 2019 Patients in Earlier Stages Exhaled Millions of Severe Acute Respiratory Syndrome Coronavirus 2 Per Hour.

Coronavirus disease 2019 (COVID-19) patients exhaled millions of severe acute respiratory syndrome coronavirus 2 RNA copies per hour, which plays an important role in COVID-19 transmission. Exhaled breath had a higher positive rate (26.9%, n = 52) than surface (5.4%, n = 242) and air (3.8%, n = 26) samples.

Breath-, air- and surface-borne SARS-CoV-2 in hospitals.

The COVID-19 pandemic has brought an unprecedented crisis to the global health sector. When discharging COVID-19 patients in accordance with throat or nasal swab protocols using RT-PCR, the potential risk of reintroducing the infection source to humans and the environment must be resolved. Here, 14 patients including 10 COVID-19 subjects were recruited; exhaled breath condensate (EBC), air samples and surface swabs were collected and analyzed for SARS-CoV-2 using reverse transcription-polymerase chain reaction (RT-PCR) in four hospitals with applied natural ventilation and disinfection practices in Wuhan. Here we discovered that 22.2% of COVID-19 patients (n = 9), who were ready for hospital discharge based on current guidelines, had SARS-CoV-2 in their exhaled breath (~105 RNA copies/m3). Although fewer surface swabs (3.1%, n = 318) tested positive, medical equipment such as face shield frequently contacted/used by healthcare workers and the work shift floor were contaminated by SARS-CoV-2 (3-8 viruses/cm2). Three of the air samples (n = 44) including those collected using a robot-assisted sampler were detected positive by a digital PCR with a concentration level of 9-219 viruses/m3. RT-PCR diagnosis using throat swab specimens had a failure rate of more than 22% in safely discharging COVID-19 patients who were otherwise still exhaling the SARS-CoV-2 by a rate of estimated ~1400 RNA copies per minute into the air. Direct surface contact might not represent a major transmission route, and lower positive rate of air sample (6.8%) was likely due to natural ventilation (1.6-3.3 m/s) and regular disinfection practices. While there is a critical need for strengthening hospital discharge standards in preventing re-emergence of COVID-19 spread, use of breath sample as a supplement specimen could further guard the hospital discharge to ensure the safety of the public and minimize the pandemic re-emergence risk.

Rats Sniff Off Toxic Air.

Breathing air is a fundamental human need, yet its safety, when challenged by various harmful or lethal substances, is often not properly guarded. For example, air toxicity is currently monitored only for a single or a limited number of known toxicants, thus failing to warn against possible hazardous air fully. Here, we discovered that, within minutes, living rats emitted distinctive profiles of volatile organic compounds (VOCs) via breath when exposed to various airborne toxicants such as endotoxin, O3, ricin, and CO2. Compared to background indoor air, when exposed to ricin or endotoxin aerosols, breath-borne VOC levels, especially that of carbon disulfide, were shown to decrease, while their elevated levels were observed for exposure to O3 and CO2. A clear contrast in breath-borne VOC profiles of rats exposed to different toxicants was observed with a statistical significance. Differences in microRNA regulations such as miR-33, miR-146a, and miR-155 from rats' blood samples revealed different mechanisms used by rats in combating different air toxicant challenges. Similar to dogs, rats were found here to be able to sniff off toxic air by releasing a specific breath-borne VOC profile. The discovered science opens a new arena for online monitoring of air toxicity and health effects of pollutants.

Ambient PM Toxicity Is Correlated with Expression Levels of Specific MicroRNAs.

Uncertainties regarding optimized air pollution control remain as the underlying mechanisms of city-specific ambient particulate matter (PM)-induced health effects are unknown. Here, water-soluble extracts of PMs collected from four global cities via automobile air-conditioning filters were consecutively injected three times by an amount of 1, 2, and 2 mg into the blood circulation of Wistar rats after filtration by a 0.45 µm pore size membrane. Acute health effects, such as immune and inflammatory responses and hemorrhage in alveoli, were observed right after the PM extraction injection. Significant differences between cities in biomarker tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) levels were detected following the second and third PM injections. Rats' inflammatory responses varied substantially with the injections of city-specific PMs. Repeated PM extract exposure rendered the rats more vulnerable to subsequent challenges, and downregulation of certain microRNAs was observed in rats. Among the studied miRNAs, miR-125b, and miR-21 were most sensitive to the PM exposure, exhibiting a negative dose-response-type relationship with a source-specific PM (oxidative potential) toxicity (r2 = 0.63 and 0.57; p-values < 0.05). The results indicated that city-specific PMs could induce different health effects by selectively regulating different miRNAs, and that certain microRNAs, e.g., miR-125b and miR-21, may be externally mediated to neutralize PM-related health damages.

Radical Formation by Fine Particulate Matter Associated with Highly Oxygenated Molecules.

Highly oxygenated molecules (HOMs) play an important role in the formation and evolution of secondary organic aerosols (SOA). However, the abundance of HOMs in different environments and their relation to the oxidative potential of fine particulate matter (PM) are largely unknown. Here, we investigated the relative HOM abundance and radical yield of laboratory-generated SOA and fine PM in ambient air ranging from remote forest areas to highly polluted megacities. By electron paramagnetic resonance and mass spectrometric investigations, we found that the relative abundance of HOMs, especially the dimeric and low-volatility types, in ambient fine PM was positively correlated with the formation of radicals in aqueous PM extracts. SOA from photooxidation of isoprene, ozonolysis of α- and ß-pinene, and fine PM from tropical (central Amazon) and boreal (Hyytiälä, Finland) forests exhibited a higher HOM abundance and radical yield than SOA from photooxidation of naphthalene and fine PM from urban sites (Beijing, Guangzhou, Mainz, Shanghai, and Xi'an), confirming that HOMs are important constituents of biogenic SOA to generate radicals. Our study provides new insights into the chemical relationship of HOM abundance, composition, and sources with the yield of radicals by laboratory and ambient aerosols, enabling better quantification of the component-specific contribution of source- or site-specific fine PM to its climate and health effects.

Automated in Vivo Nanosensing of Breath-Borne Protein Biomarkers.

Toxicology and bedside medical condition monitoring is often desired to be both ultrasensitive and noninvasive. However, current biomarker analyses for these purposes are mostly offline and fail to detect low marker quantities. Here, we report a system called dLABer (detection of living animal's exhaled breath biomarker) that integrates living rats, breath sampling, microfluidics, and biosensors for the automated tracking of breath-borne biomarkers. Our data show that dLABer could selectively detect (online) and report differences (of up to 103-fold) in the levels of inflammation agent interleukin-6 (IL-6) exhaled by rats injected with different ambient particulate matter (PM). The dLABer system was further shown to have an up to 104 higher signal-to-noise ratio than that of the enzyme-linked immunosorbent assay (ELISA) when analyzing the same breath samples. In addition, both blood-borne IL-6 levels analyzed via ELISA in rats injected with different PM extracts and PM toxicity determined by a dithiothreitol (DTT) assay agreed well with those determined by the dLABer system. Video recordings further verified that rats exposed to PM with higher toxicity (according to a DTT assay and as revealed by dLABer) appeared to be less physically active. All the data presented here suggest that the dLABer system is capable of real-time, noninvasive monitoring of breath-borne biomarkers with ultrasensitivity. The dLABer system is expected to revolutionize pollutant health effect studies and bedside disease diagnosis as well as physiological condition monitoring at the single-protein level.

Size-Resolved Endotoxin and Oxidative Potential of Ambient Particles in Beijing and Zürich.

PM2.5 pollution has become a global health concern, however its size-resolved health impact remains to be poorly elucidated. Here, ambient particulate matter (PM) were collected into 13 different size ranges (10 nm to 18 µm) and the mass, metal, endotoxin distributions, and related oxidative potential were investigated in two regions (Zürich, Switzerland and Beijing, China). Results showed that the two regions had remarkably different PM distribution patterns. Swiss urban samples had a mode around 40 nm with 23.3% of total PM mass, while Chinese samples featured two modes around 0.75 and 4.23 µm with 13.8-18.6% and 13.7-20.4% of total PM mass, respectively. Two peaks for endotoxin at 40-100 nm and 1-4 µm were observed in different regions. For PM-borne metals, Chinese samples had 67.6-100% of total Cd, As, and Pb in the size range of 0.1-1 µm, and Swiss samples had similar distributions of Cd and Pb but much lower total metals than Chinese samples. The PM oxidative potential varied greatly with sizes for different regions. Accordingly, the current practice, i.e., sole use of the mass concentration, could lead to inadequate health protection for one region, but unnecessary economic costs for another without achieving significant extra health benefits.

Global Survey of Antibiotic Resistance Genes in Air.

Despite its emerging significant public health concern, the presence of antibiotic resistance genes (ARGs) in urban air has not received significant attention. Here, we profiled relative abundances (as a fraction, normalized by 16S rRNA gene) of 30 ARG subtypes resistant to seven common classes of antibiotics, which are quinolones, ß-lactams, macrolides, tetracyclines, sulfonamides, aminoglycosides, and vancomycins, in ambient total particulate matter (PM) using a novel protocol across 19 world cities. In addition, their longitudinal changes in PM2.5 samples in Xi'an, China as an example were also studied. Geographically, the ARGs were detected to vary by nearly 100-fold in their abundances, for example, from 0.07 (Bandung, Indonesia) to 5.6 (San Francisco, USA). The ß-lactam resistance gene blaTEM was found to be most abundant, seconded by quinolone resistance gene qepA; and their corresponding relative abundances have increased by 178% and 26%, respectively, from 2004 to 2014 in Xi'an. Independent of cities, gene network analysis indicates that airborne ARGs were differentially contributed by bacterial taxa. Results here reveal that urban air is being polluted by ARGs, and different cities are challenged with varying health risks associated with airborne ARG exposure. This work highlights the threat of urban airborne transmission of ARGs and the need of redefining our current air quality standards in terms with public health.

Reprint of bioaerosol: A bridge and opportunity for many scientific research fields.

Bioaerosol is a concept that is used to describe all biological materials suspended in the air, including bacteria, fungi, viruses, pollen, and their derivatives such as allergens, endotoxin, mycotoxins and etc. In some studies, primary biological aerosol particle (PBAP) is also coined to refer to intact microbes in the air. Bioaerosol is a multidisciplinary research subject, involving many different fields such as microbiology, mechanical engineering, air pollution, medical science, epidemiology, immunological science, biochemistry, physics, nanotechnologies and etc. The bioaerosol field has undergone about 200 years' research history since 1833 when mold spores were first detected in the air by Charles Darwin on the Cape Verde Islands. In recent decades, there has been a research boom in bioaerosol field, thus triggering many outstanding research opportunities. Visible progress has already been made in understanding bioaerosol roles in human health, atmospheric and ecological impacts as well as their respective technologies: bioaerosol capture, monitoring and also inactivation. Most recently, researchers from different fields start to bridge together for solving bioaerosol challenges and addressing key scientific problems, e.g., bioaerosol spread, real-time detection, indoor microbes, human bioaerosol emissions, and bio-defense. Toward this effort, a "Bioaerosol Xiangshan Science Conference-the 600th" has been successfully held in the summer in Beijing, China. A total of 47 scientists and funding agency officials including leading bioaerosol experts from overseas were invited and two-day long extensive discussions on bioaerosol progress and problems were carried out. Future bioaerosol directions have been outlined by the attendees during the conference. Some of the participants have also contributed to this bioaerosol special issue. This special issue consists of a total of 20 bioaerosol articles from eight countries including one review, and contributes to the advances in bioaerosol emission, transmission, health effects, ambient bioaerosols, method development and instrumentation, and control. Through this special issue, the bioaerosol community has obtained a better understanding of bioaerosol health risks and developed the corresponding strategies to confront the threats. This special issue might serve as a starting point to not only link bioaerosol scientists from different continents, but also bring together people from various fields yet with an interest in bioaerosol to collectively advance the field further.

A high-flow portable biological aerosol trap (HighBioTrap) for rapid microbial detection.

Bioaerosols exposure can lead to many adverse health effects and even result in death if highly infectious agents involved. Apparently, there is a great need for rapid detection of bioaerosols, for which air sampling often is the first critical step. However, currently available samplers often either require an external power and/or with low sampling flow rate, thus falling short of providing a practical solution when response time is of great concern. Here, we have designed and evaluated a new portable high volume bioaerosol sampler named as HighBioTrap through optimizing its operating parameters. The sampler was operated at a sampling flow rate of 1200 L/min, with an impaction velocity of about 10.2 m/s (S/W = 1.5, T/W = 1), while the weight of the sampler is about 1.9 kg. The performances of the HighBioTrap sampler were tested both in lab controlled and natural environments (outdoor and indoor environments in a university building) along with the reference sampler-the BioStage impactor using aerosolized Polystyrene (PS) uniform microspheres of various sizes, aerosolized bacteria and also ambient air particles. The microbial community structures of collected culturable bacterial aerosol particles both by the HighBioTrap and the BioStage impactor in the natural environments were analyzed using gene sequence method. Experimental results with PS particles showed the HighBioTrap has a cutoff size of ~ 2 µm. The widely used impactor design equation was found to be not applicable for predicting the performance of the HighBioTrap due to its large Reynolds number. When sampling aerosolized individual Pseudomonas fluorescens and Bacillus subtilis bacterial particles, the HighBioTrap had physical collection efficiencies of 10% and 20%, respectively. Despite the higher desiccation effects introduced by higher flow rate, the HighBioTrap was shown to obtain a higher microbial diversity than the BioStage impactor for both in outdoor and indoor environments given the same sampling time (p < 0.01). Our data also showed that most of the desiccation effects might have occurred between 3 and 5 min of the sampling and an impaction velocity of around 10 m/s might be a close-to-optimal impaction velocity for collecting most environmental bacterial aerosols while maximally preserving their culturability. This work contributes to our understanding of microbial sampling stress (impaction velocity and sampling time), while developing a portable high volume sampler. The HighBioTrap sampler could find its great efficiencies in qualitative microbial aerosol detection and analysis, such as investigation of microbial aerosol diversity for a particular environment, or when the low level of pathogens is present and detection time is of great concern.

Bacterial pathogens were detected from human exhaled breath using a novel protocol.

It is generally believed that influenza outbreak is associated with breath-borne transmission of viruses, however relevant evidence is little for that of respiratory bacterial infections. On another front, point-of-care infection diagnostic methods at the bedside are significantly lacking. Here, we used a newly developed protocol of integrating an exhaled breath condensate (EBC) collection device (PKU BioScreen) and Loop Mediated Isothermal Amplification (LAMP) to investigate what bacterial pathogens can be directly exhaled out from humans. Exhaled breath condensates were collected from human subjects with respiratory infection symptoms at Peking University 3rd hospital using the BioScreen. The screened bacterial pathogens included Streptococcus pneumoniae, Staphylococcus aureus, Methicillin-resistant Stphylococcus aureus (MRSA), Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, Haemophilus influenzae, Legionella pneumophila, Mycoplasma Pneumonia, Chlamydia pneumonia, and Mycobacterium tuberculosis. The results were further compared and validated using throat swabs from the same patients by a PCR method. Here, human bacterial pathogens such as H. influenzae, P. aeruginosa, E. coli, S. aureus and MRSA were detected in exhaled breath using the developed protocol that integrates the EBC collection and LAMP. For the patients recruited from the hospital, seven types of pathogens were detected from 36.5% of them, and for the remaining subjects none of those screened bacterial pathogens was detected. Importantly, some super resistant bacteria such as MRSA were detected from the exhaled breath, suggesting that breathing might be also an important bacterial transmission route. Results from throat swabs showed that 36.2% of the subjects were found to be infected with H. influenzae, P. aeruginosa, E. coli, S. maltophilia, S. aureus and MRSA. For the EBC samples, 33.3% were found to be infected with MRSA, E. coli and P. aeruginosa. Depending on the initial pathogen load in the sample, the entire protocol (EBC-LAMP) only takes 20-60 min to complete for a respiratory infection diagnosis. For different detection methods and pathogens, the agreements between the EBC and throat swabs from the same patients were found to range from 35% to 65%. Here, we have detected several bacterial pathogens including MRSA from exhaled breath, and the developed protocol could be very useful for the bedside pathogen screening particularly in remote areas where resources are significantly limited or prohibited.