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1.  Development and Performance Evaluation of an Exhaled-Breath Bioaerosol Collector for Influenza Virus 
The importance of the aerosol mode for transmission of influenza is unknown. Understanding the role of aerosols is essential to developing public health interventions such as the use of surgical masks as a source control to prevent the release of infectious aerosols. Little information is available on the number and size of particles generated by infected persons, which is partly due to the limitations of conventional air samplers, which do not efficiently capture fine particles or maintain microorganism viability. We designed and built a new sampler, called the G-II, that collects exhaled breath particles that can be used in infectivity analyses. The G-II allows test subjects to perform various respiratory maneuvers (i.e. tidal breathing, coughing, and talking) and allows subjects to wear a mask or respirator during testing. A conventional slit impactor collects particles > 5.0 μm. Condensation of water vapor is used to grow remaining particles, including fine particles, to a size large enough to be efficiently collected by a 1.0 μm slit impactor and be deposited into a buffer-containing collector. We evaluated the G-II for fine particle collection efficiency with inert particle aerosols and evaluated infective virus collection using influenza A virus aerosols. Testing results showed greater than 85% collection efficiency for particles greater than 50nm and influenza virus collection comparable with a reference SKC BioSampler®. The new design will enable determination of exhaled infectious virus generation rate and evaluate control strategies such as wearing a surgical type mask to prevent the release of viruses from infected persons.
doi:10.1080/02786826.2012.762973
PMCID: PMC3570155  PMID: 23418400
2.  Measurements of Airborne Influenza Virus in Aerosol Particles from Human Coughs 
PLoS ONE  2010;5(11):e15100.
Influenza is thought to be communicated from person to person by multiple pathways. However, the relative importance of different routes of influenza transmission is unclear. To better understand the potential for the airborne spread of influenza, we measured the amount and size of aerosol particles containing influenza virus that were produced by coughing. Subjects were recruited from patients presenting at a student health clinic with influenza-like symptoms. Nasopharyngeal swabs were collected from the volunteers and they were asked to cough three times into a spirometer. After each cough, the cough-generated aerosol was collected using a NIOSH two-stage bioaerosol cyclone sampler or an SKC BioSampler. The amount of influenza viral RNA contained in the samplers was analyzed using quantitative real-time reverse-transcription PCR (qPCR) targeting the matrix gene M1. For half of the subjects, viral plaque assays were performed on the nasopharyngeal swabs and cough aerosol samples to determine if viable virus was present. Fifty-eight subjects were tested, of whom 47 were positive for influenza virus by qPCR. Influenza viral RNA was detected in coughs from 38 of these subjects (81%). Thirty-five percent of the influenza RNA was contained in particles >4 µm in aerodynamic diameter, while 23% was in particles 1 to 4 µm and 42% in particles <1 µm. Viable influenza virus was detected in the cough aerosols from 2 of 21 subjects with influenza. These results show that coughing by influenza patients emits aerosol particles containing influenza virus and that much of the viral RNA is contained within particles in the respirable size range. The results support the idea that the airborne route may be a pathway for influenza transmission, especially in the immediate vicinity of an influenza patient. Further research is needed on the viability of airborne influenza viruses and the risk of transmission.
doi:10.1371/journal.pone.0015100
PMCID: PMC2994911  PMID: 21152051
3.  The generation of diesel exhaust particle aerosols from a bulk source in an aerodynamic size range similar to atmospheric particles 
The influence of diesel exhaust particles (DEP) on the lungs and heart is currently a topic of great interest in inhalation toxicology. Epidemiological data and animal studies have implicated airborne particulate matter and DEP in increased morbidity and mortality due to a number of cardiopulmonary diseases including asthma, chronic obstructive pulmonary disorder, and lung cancer. The pathogeneses of these diseases are being studied using animal models and cell culture techniques. Real-time exposures to freshly combusted diesel fuel are complex and require significant infrastructure including engine operations, dilution air, and monitoring and control of gases. A method of generating DEP aerosols from a bulk source in an aerodynamic size range similar to atmospheric DEP would be a desirable and useful alternative. Metered dose inhaler technology was adopted to generate aerosols from suspensions of DEP in the propellant hydrofluoroalkane 134a. Inertial impaction data indicated that the particle size distributions of the generated aerosols were trimodal, with count median aerodynamic diameters less than 100 nm. Scanning electron microscopy of deposited particles showed tightly aggregated particles, as would be expected from an evaporative process. Chemical analysis indicated that there were no major changes in the mass proportion of 2 specific aromatic hydrocarbons (benzo[a]pyrene and benzo[k]fluoranthene) in the particles resulting from the aerosolization process.
PMCID: PMC2636590  PMID: 19337412
diesel exhaust particles; aerosol; inhalation toxicology
4.  Development of a Sampler for Total Aerosol Deposition in the Human Respiratory Tract 
Annals of Occupational Hygiene  2009;53(7):731-738.
Studies that seek to associate reduced human health with exposure to occupational and environmental aerosols are often hampered by limitations in the exposure assessment process. One limitation involves the measured exposure metric itself. Current methods for personal exposure assessment are designed to estimate the aspiration of aerosol into the human body. Since a large proportion of inhaled aerosol is subsequently exhaled, a portion of the aspirated aerosol will not contribute to the dose. This leads to variable exposure misclassification (for heterogenous exposures) and increased uncertainty in health effect associations. Alternatively, a metric for respiratory deposition would provide a more physiologically relevant estimate of risk. To address this challenge, we have developed a method to estimate the deposition of aerosol in the human respiratory tract using a sampler engineered from polyurethane foam. Using a semi-empirical model based on inertial, gravitational, and diffusional particle deposition, a foam was engineered to mimic aerosol total deposition in the human respiratory tract. The sampler is comprised of commercially available foam with fiber diameter = 49.5 μm (equivalent to industry standard 100 PPI foam) of 8 cm thickness operating at a face velocity of 1.3 m s−1. Additionally, the foam sampler yields a relatively low-pressure drop, independent of aerosol loading, providing uniform particle collection efficiency over time.
doi:10.1093/annhyg/mep053
PMCID: PMC2913619  PMID: 19638392
aerosols; exposure estimation; gravimetric analysis
5.  Detection and Isolation of Airborne Influenza A H3N2 Virus Using a Sioutas Personal Cascade Impactor Sampler 
The air we breathe contains microorganisms that can cause infectious respiratory diseases. After two occupants of an apartment were diagnosed with influenza in February of 2013, efforts were made to detect and isolate airborne influenza virus using two different types of active air samplers: a Sioutas Personal Cascade Impactor Sampler (PCIS) and an SKC BioSampler. The PCIS collects size-fractionated particles by impaction on polytetrafluoroethylene filters, whereas the SKC BioSampler collects airborne particles in liquid media. Influenza H3N2 virus was collected by both types of air samplers. The PCIS collected a range of particle sizes containing influenza virus near one of the sick individuals but only ultrafine particles when the samplers were positioned farther away. Viable virus was present in the liquid collection media of the SKC BioSampler and some PCIS filters. These findings suggest that influenza patients produce ultrafine aerosol particles that contain viable virus.
doi:10.1155/2013/656825
PMCID: PMC3810434  PMID: 24224087
6.  Performance of High Flow Rate Samplers for Respirable Particle Collection 
Annals of Occupational Hygiene  2010;54(6):697-709.
The American Conference of Governmental Industrial hygienists (ACGIH) lowered the threshold limit value (TLV) for respirable crystalline silica (RCS) exposure from 0.05 to 0.025 mg m−3 in 2006. For a working environment with an airborne dust concentration near this lowered TLV, the sample collected with current standard respirable aerosol samplers might not provide enough RCS for quantitative analysis. Adopting high flow rate sampling devices for respirable dust containing silica may provide a sufficient amount of RCS to be above the limit of quantification even for samples collected for less than full shift. The performances of three high flow rate respirable samplers (CIP10-R, GK2.69, and FSP10) have been evaluated in this study. Eleven different sizes of monodisperse aerosols of ammonium fluorescein were generated with a vibrating orifice aerosol generator in a calm air chamber in order to determine the sampling efficiency of each sampler. Aluminum oxide particles generated by a fluidized bed aerosol generator were used to test (i) the uniformity of a modified calm air chamber, (ii) the effect of loading on the sampling efficiency, and (iii) the performance of dust collection compared to lower flow rate cyclones in common use in the USA (10-mm nylon and Higgins–Dewell cyclones). The coefficient of variation for eight simultaneous samples in the modified calm air chamber ranged from 1.9 to 6.1% for triplicate measures of three different aerosols. The 50% cutoff size (50dae) of the high flow rate samplers operated at the flow rates recommended by manufacturers were determined as 4.7, 4.1, and 4.8 μm for CIP10-R, GK2.69, and FSP10, respectively. The mass concentration ratio of the high flow rate samplers to the low flow rate cyclones decreased with decreasing mass median aerodynamic diameter (MMAD) and high flow rate samplers collected more dust than low flow rate samplers by a range of 2–11 times based on gravimetric analysis. Dust loading inside the high flow rate samplers does not appear to affect the particle separation in either FSP10 or GK2.69. The high flow rate samplers overestimated compared to the International Standards Organization/Comité Européen de Normalisation/ACGIH respirable convention [up to 40% at large MMAD (27.5 μm)] and could provide overestimated exposure data with the current flow rates. However, both cyclones appeared to be able to provide relatively unbiased assessments of RCS when their flow rates were adjusted.
doi:10.1093/annhyg/meq050
PMCID: PMC2918491  PMID: 20660144
calm air chamber; CIP10-R; FSP10; GK2.69; high flow rate sampler; respirable fraction; sampling efficiency
7.  Aerosol Monitoring during Carbon Nanofiber Production: Mobile Direct-Reading Sampling 
Annals of Occupational Hygiene  2010;54(5):514-531.
Detailed investigations were conducted at a facility that manufactures and processes carbon nanofibers (CNFs). Presented research summarizes the direct-reading monitoring aspects of the study. A mobile aerosol sampling platform, equipped with an aerosol instrument array, was used to characterize emissions at different locations within the facility. Particle number, respirable mass, active surface area, and photoelectric response were monitored with a condensation particle counter (CPC), a photometer, a diffusion charger, and a photoelectric aerosol sensor, respectively. CO and CO2 were additionally monitored. Combined simultaneous monitoring of these metrics can be utilized to determine source and relative contribution of airborne particles (CNFs and others) within a workplace. Elevated particle number concentrations, up to 1.15 × 106 cm−3, were found within the facility but were not due to CNFs. Ultrafine particle emissions, released during thermal treatment of CNFs, were primarily responsible. In contrast, transient increases in respirable particle mass concentration, with a maximum of 1.1 mg m−3, were due to CNF release through uncontrolled transfer and bagging. Of the applied metrics, our findings suggest that particle mass was probably the most useful and practical metric for monitoring CNF emissions in this facility. Through chemical means, CNFs may be selectively distinguished from other workplace contaminants (Birch et al., in preparation), and for direct-reading monitoring applications, the photometer was found to provide a reasonable estimate of respirable CNF mass concentration. Particle size distribution measurements were conducted with an electrical low-pressure impactor and a fast particle size spectrometer. Results suggest that the dominant CNF mode by particle number lies between 200 and 250 nm for both aerodynamic and mobility equivalent diameters. Significant emissions of CO were also evident in this facility. Exposure control recommendations were described for processes as required.
doi:10.1093/annhyg/meq015
PMCID: PMC2900095  PMID: 20447936
carbon monoxide; emissions; exposure; exposure controls; nanofibers; nanomaterial; nanoparticle; nanotubes; occupational; ultrafine; workplace
8.  Characterization of Red Tide Aerosol on the Texas Coast 
Harmful algae  2005;4(1):87-94.
The Gulf of Mexico red tide, caused by the dinoflagellate Karenia brevis (= Gymnodinium breve), occurs almost annually and has adverse economic and health effects. Exposure of people to sea spray containing aerosolized brevetoxins (PbTxs, polyether brevetoxins produced by K. brevis) causes irritation of the eyes, nose, and throat. Anecdotal reports suggest that exposed individuals can experience respiratory irritation and exacerbation of existing respiratory illnesses. There has been no systematic study of human exposure to red tide aerosols. In the fall of 2000, during a red tide episode on the Gulf Coast near Corpus Christi, Texas, we sampled at the Marine Science Institute (MSI) at Port Aransas on 25 October. Between 26–27 October we sampled at the Texas State Aquarium (TSA) near Corpus Christi. Two Hi-Vol samplers equipped with a filter and a five-stage impactor gave low concentrations of PbTxs, requiring us to develop methods to improve the minimum detection limit. An LC/MS/MS technique was used combining an HPLC and the API 365 MS/MS. PbTx-2 and PbTx-3 were detected at the TSA sampling location; however, PbTx was not detected in the samples from the MSI. The concentration of PbTx-2 was 1.5–4.9 ng m–3 but was much lower for PbTx-3. The ratio of PbTx-2 to PbTx-3 was 8.7 ± 5.2. During the highest exposure period (26–27 October), PbTx-6 was also detected. No one reported respiratory symptoms at the MSI, whereas at the TSA, several field study workers reported symptoms including nose and throat irritation, and itchy skin. A high-volume impactor was used to aerodynamically classify the particles into different size fractions. PbTx-2 was detected in all samples taken at the TSA; however, PbTx-3 was detected only between 26–27 October when the PbTx concentration was high. The mass median aerodynamic diameter (MMAD) was 7–9 mm with a relatively narrow size range (geometric standard deviation [GSD] about 1.6). In this study, much lower airborne concentrations of PbTx, 1.6–6.7 ng m–3 were reported, along with a few incidents of upper respiratory symptoms. Although the number of seven workers was too small for statistical analysis, the reported symptoms were consistent with no to low exposure at the MSI and detectable exposures at the TSA. This suggests that at lower environmental concentrations of about 2–7 ng m–3,exposure to PbTx could result in upper respiratory symptoms. This is consistent with the particle size measurement.
doi:10.1016/j.hal.2003.12.002
PMCID: PMC2845976  PMID: 20352032
9.  The Effect of Nonideal Cascade Impactor Stage Collection Efficiency Curves on the Interpretation of the Size of Inhaler-Generated Aerosols 
AAPS PharmSciTech  2013;14(2):497-510.
Cascade impactors, operating on the principle of inertial size separation in (ideally) laminar flow, are used to determine aerodynamic particle size distributions (APSDs) of orally inhaled product (OIP) aerosols because aerodynamic diameter can be related to respiratory tract deposition. Each stage is assumed typically to be an ideal size fractionator. Thus, all particles larger than a certain size are considered collected and all finer particles are treated as penetrating to the next stage (a step function stage efficiency curve). In reality, the collection efficiency of a stage smoothly increases with particle size as an “S-shaped” curve, from approximately 0% to 100%. Consequently, in some cases substantial overlap occurs between neighboring stages. The potential for bias associated with the step-function assumption has been explored, taking full resolution and two-stage abbreviated forms of the Andersen eight-stage nonviable impactor (ACI) and the next-generation pharmaceutical impactor (NGI) as example apparatuses. The behavior of unimodal, log-normal APSDs typical of OIP-generated aerosols has been investigated, comparing known input values to calculated values of central tendency (mass median aerodynamic diameter) and spread (geometric standard deviation, GSD). These calculations show that the error introduced by the step change assumption is larger for the ACI than for the NGI. However, the error is sufficiently small to be inconsequential unless the APSD in nearly monodisperse (GSD ≤1.2), a condition that is unlikely to occur with realistic OIPs. Account may need to be taken of this source of bias only for the most accurate work with abbreviated ACI systems.
Electronic supplementary material
The online version of this article (doi:10.1208/s12249-013-9936-2) contains supplementary material, which is available to authorized users.
doi:10.1208/s12249-013-9936-2
PMCID: PMC3666003  PMID: 23508617
cascade impactor; inhaler aerosol; inhaler testing; size distribution
10.  Particle Deposition in a Child Respiratory Tract Model: In Vivo Regional Deposition of Fine and Ultrafine Aerosols in Baboons 
PLoS ONE  2014;9(4):e95456.
To relate exposure to adverse health effects, it is necessary to know where particles in the submicron range deposit in the respiratory tract. The possibly higher vulnerability of children requires specific inhalation studies. However, radio-aerosol deposition experiments involving children are rare because of ethical restrictions related to radiation exposure. Thus, an in vivo study was conducted using three baboons as a child respiratory tract model to assess regional deposition patterns (thoracic region vs. extrathoracic region) of radioactive polydisperse aerosols ([d16–d84], equal to [0.15 µm–0.5 µm], [0.25 µm–1 µm], or [1 µm–9 µm]). Results clearly demonstrated that aerosol deposition within the thoracic region and the extrathoraic region varied substantially according to particle size. High deposition in the extrathoracic region was observed for the [1 µm–9 µm] aerosol (72%±17%). The [0.15 µm–0.5 µm] aerosol was associated almost exclusively with thoracic region deposition (84%±4%). Airborne particles in the range of [0.25 µm–1 µm] showed an intermediate deposition pattern, with 49%±8% in the extrathoracic region and 51%±8% in the thoracic region. Finally, comparison of baboon and human inhalation experiments for the [1 µm–9 µm] aerosol showed similar regional deposition, leading to the conclusion that regional deposition is species-independent for this airborne particle sizes.
doi:10.1371/journal.pone.0095456
PMCID: PMC4005734  PMID: 24787744
11.  Nanomaterial inhalation exposure from nanotechnology-based cosmetic powders: a quantitative assessment 
In this study we quantified exposures to airborne particles ranging from 14 nm to 20 µm due to the use of nanotechnology-based cosmetic powders. Three nanotechnology-based and three regular cosmetic powders were realistically applied to a mannequin’s face while measuring the concentration and size distribution of inhaled aerosol particles. Using these data we calculated that the highest inhaled particle mass was in the coarse aerosol fraction (2.5–10 µm), while particles <100 nm made minimal contribution to the inhaled particle mass. For all powders, 85–93 % of aerosol deposition occurred in the head airways, while <10 % deposited in the alveolar and <5 % in the tracheobronchial regions. Electron microscopy data suggest that nanomaterials were likely distributed as agglomerates across the entire investigated aerosol size range (14 nm–20 µm). Thus, investigation of nanoparticle health effects should consider not only the alveolar region, but also other respiratory system regions where substantial nanomaterial deposition during the actual nanotechnology-based product use would occur.
doi:10.1007/s11051-012-1229-2
PMCID: PMC3500971  PMID: 23175627
Nanoaerosol; Consumer products; Nanoparticles; Personal exposure; Safety of nanotechnology
12.  Characterization of Exposures to Airborne Nanoscale Particles During Friction Stir Welding of Aluminum 
Annals of Occupational Hygiene  2010;54(5):486-503.
Friction stir welding (FSW) is considered one of the most significant developments in joining technology over the last half century. Its industrial applications are growing steadily and so are the number of workers using this technology. To date, there are no reports on airborne exposures during FSW. The objective of this study was to investigate possible emissions of nanoscale (<100 nm) and fine (<1 μm) aerosols during FSW of two aluminum alloys in a laboratory setting and characterize their physicochemical composition. Several instruments measured size distributions (5 nm to 20 μm) with 1-s resolution, lung deposited surface areas, and PM2.5 concentrations at the source and at the breathing zone (BZ). A wide range aerosol sampling system positioned at the BZ collected integrated samples in 12 stages (2 nm to 20 μm) that were analyzed for several metals using inductively coupled plasma mass spectrometry. Airborne aerosol was directly collected onto several transmission electron microscope grids and the morphology and chemical composition of collected particles were characterized extensively. FSW generates high concentrations of ultrafine and submicrometer particles. The size distribution was bimodal, with maxima at ∼30 and ∼550 nm. The mean total particle number concentration at the 30 nm peak was relatively stable at ∼4.0 × 105 particles cm−3, whereas the arithmetic mean counts at the 550 nm peak varied between 1500 and 7200 particles cm−3, depending on the test conditions. The BZ concentrations were lower than the source concentrations by 10–100 times at their respective peak maxima and showed higher variability. The daylong average metal-specific concentrations were 2.0 (Zn), 1.4 (Al), and 0.24 (Fe) μg m−3; the estimated average peak concentrations were an order of magnitude higher. Potential for significant exposures to fine and ultrafine aerosols, particularly of Al, Fe, and Zn, during FSW may exist, especially in larger scale industrial operations.
doi:10.1093/annhyg/meq037
PMCID: PMC2913760  PMID: 20453001
aluminum; friction stir welding; nanoparticle exposures; occupational safety and health; size distribution; zinc
13.  Hygroscopic and Chemical Properties of Aerosols collected near a Copper Smelter: Implications for Public and Environmental Health 
Environmental science & technology  2012;46(17):9473-9480.
Particulate matter emissions near active copper smelters and mine tailings in the southwestern United States pose a potential threat to nearby environments owing to toxic species that can be inhaled and deposited in various regions of the body depending on the composition and size of the particles, which are linked by particle hygroscopic properties. This study reports the first simultaneous measurements of size-resolved chemical and hygroscopic properties of particles next to an active copper smelter and mine tailings by the towns of Hayden and Winkelman in southern Arizona. Size-resolved particulate matter samples collected near an active copper smelter were examined with inductively coupled plasma mass spectrometry, ion chromatography, and a humidified tandem differential mobility analyzer. Aerosol particles collected at the measurement site are enriched in metals and metalloids (e.g. arsenic, lead, and cadmium) and water-uptake measurements of aqueous extracts of collected samples indicate that the particle diameter range of particles most enriched with these species (0.18–0.55 µm) overlaps with the most hygroscopic mode at a relative humidity of 90% (0.10–0.32 µm). These measurements have implications for public health, microphysical effects of aerosols, and regional impacts owing to the transport and deposition of contaminated aerosol particles.
doi:10.1021/es302275k
PMCID: PMC3435440  PMID: 22852879
Particulate Matter; Soil Dust; Hygroscopicity; Smelter; Arizona; Mine Tailings; Arsenic; Lead; Oxalate; Organic Acid
14.  Metal and Metalloid Contaminants in Atmospheric Aerosols from Mining Operations 
Water, air, and soil pollution  2011;221(1-4):145-157.
Mining operations are potential sources of airborne metal and metalloid contaminants through both direct smelter emissions and wind erosion of mine tailings. The warmer, drier conditions predicted for the Southwestern US by climate models may make contaminated atmospheric dust and aerosols increasingly important, with potential deleterious effects on human health and ecology. Fine particulates such as those resulting from smelting operations may disperse more readily into the environment than coarser tailings dust. Fine particles also penetrate more deeply into the human respiratory system, and may become more bioavailable due to their high specific surface area. In this work, we report the size-fractionated chemical characterization of atmospheric aerosols sampled over a period of a year near an active mining and smelting site in Arizona. Aerosols were characterized with a 10-stage (0.054 to 18 μm aerodynamic diameter) multiple orifice uniform deposit impactor (MOUDI), a scanning mobility particle sizer (SMPS), and a total suspended particulate (TSP) collector. The MOUDI results show that arsenic and lead concentrations follow a bimodal distribution, with maxima centered at approximately 0.3 and 7.0 μm diameter. We hypothesize that the sub-micron arsenic and lead are the product of condensation and coagulation of smelting vapors. In the coarse size, contaminants are thought to originate as aeolian dust from mine tailings and other sources. Observation of ultrafine particle number concentration (SMPS) show the highest readings when the wind comes from the general direction of the smelting operations site.
doi:10.1007/s11270-011-0777-x
PMCID: PMC3576728  PMID: 23441050
Mining Operations; MOUDI; SMPS; Aerosol; Metal Contaminants; Metalloid Contaminants
15.  Enhancing Bioaerosol Sampling by Andersen Impactors Using Mineral-Oil-Spread Agar Plate 
PLoS ONE  2013;8(2):e56896.
As a bioaerosol sampling standard, Andersen type impactor is widely used since its invention in 1950s, including the investigation of the anthrax attacks in the United States in 2001. However, its related problems such as impaction and desiccation stress as well as particle bounce have not been solved. Here, we improved its biological collection efficiencies by plating a mineral oil layer (100 µL) onto the agar plate. An Andersen six-stage sampler and a BioStage impactor were tested with mineral-oil-spread agar plates in collecting indoor and outdoor bacterial and fungal aerosols. The effects of sampling times (5, 10 and 20 min) were also studied using the BioStage impactor when sampling environmental bioaerosols as well as aerosolized Bacillus subtilis (G+) and Escherichia coli (G-). In addition, particle bounce reduction by mineral-oil-plate was also investigated using an optical particle counter (OPC). Experimental results revealed that use of mineral-oil-spread agar plate can substantially enhance culturable bioaerosol recoveries by Andersen type impactors (p-values<0.05). The recovery enhancement was shown to depend on bioaerosol size, type, sampling time and environment. In general, more enhancements (extra 20%) were observed for last stage of the Andersen six-stage samplers compared to the BioStage impactor for 10 min sampling. When sampling aerosolized B. subtilis, E. coli and environmental aerosols, the enhancement was shown to increase with increasing sampling time, ranging from 50% increase at 5 min to ∼100% at 20 min. OPC results indicated that use of mineral oil can effectively reduce the particle bounce with an average of 66% for 10 min sampling. Our work suggests that enhancements for fungal aerosols were primarily attributed to the reduced impaction stress, while for bacterial aerosols reduced impaction, desiccation and particle bounce played major roles. The developed technology can readily enhance the agar-based techniques including those high volume portable samplers for bioaerosol monitoring.
doi:10.1371/journal.pone.0056896
PMCID: PMC3584084  PMID: 23460818
16.  Bioaerosol Mass Spectrometry for Rapid Detection of Individual Airborne Mycobacterium tuberculosis H37Ra Particles 
Applied and Environmental Microbiology  2005;71(10):6086-6095.
Single-particle laser desorption/ionization time-of-flight mass spectrometry, in the form of bioaerosol mass spectrometry (BAMS), was evaluated as a rapid detector for individual airborne, micron-sized, Mycobacterium tuberculosis H37Ra particles, comprised of a single cell or a small number of clumped cells. The BAMS mass spectral signatures for aerosolized M. tuberculosis H37Ra particles were found to be distinct from M. smegmatis, Bacillus atrophaeus, and B. cereus particles, using a distinct biomarker. This is the first time a potentially unique biomarker was measured in M. tuberculosis H37Ra on a single-cell level. In addition, M. tuberculosis H37Ra and M. smegmatis were aerosolized into a bioaerosol chamber and were sampled and analyzed using BAMS, an aerodynamic particle sizer, a viable Anderson six-stage sampler, and filter cassette samplers that permitted direct counts of cells. In a background-free environment, BAMS was able to sample and detect M. tuberculosis H37Ra at airborne concentrations of >1 M. tuberculosis H37Ra-containing particles/liter of air in 20 min as determined by direct counts of filter cassette-sampled particles, and concentrations of >40 M. tuberculosis H37Ra CFU/liter of air in 1 min as determined by using viable Andersen six-stage samplers. This is a first step toward the development of a rapid, stand-alone airborne M. tuberculosis particle detector for the direct detection of M. tuberculosis bioaerosols generated by an infectious patient. Additional instrumental development is currently under way to make BAMS useful in realistic environmental and respiratory particle backgrounds expected in tuberculosis diagnostic scenarios.
doi:10.1128/AEM.71.10.6086-6095.2005
PMCID: PMC1265962  PMID: 16204525
17.  Wood Dust Sampling: Field Evaluation of Personal Samplers When Large Particles Are Present 
Annals of Occupational Hygiene  2010;55(2):180-191.
Recent recommendations for wood dust sampling include sampling according to the inhalable convention of International Organization for Standardization (ISO) 7708 (1995) Air quality—particle size fraction definitions for health-related sampling. However, a specific sampling device is not mandated, and while several samplers have laboratory performance approaching theoretical for an ‘inhalable’ sampler, the best choice of sampler for wood dust is not clear. A side-by-side field study was considered the most practical test of samplers as laboratory performance tests consider overall performance based on a wider range of particle sizes than are commonly encountered in the wood products industry. Seven companies in the wood products industry of the Southeast USA (MS, KY, AL, and WV) participated in this study. The products included hardwood flooring, engineered hardwood flooring, door skins, shutter blinds, kitchen cabinets, plywood, and veneer. The samplers selected were 37-mm closed-face cassette with ACCU-CAP™, Button, CIP10-I, GSP, and Institute of Occupational Medicine. Approximately 30 of each possible pairwise combination of samplers were collected as personal sample sets. Paired samplers of the same type were used to calculate environmental variance that was then used to determine the number of pairs of samples necessary to detect any difference at a specified level of confidence. Total valid sample number was 888 (444 valid pairs). The mass concentration of wood dust ranged from 0.02 to 195 mg m−3. Geometric mean (geometric standard deviation) and arithmetic mean (standard deviation) of wood dust were 0.98 mg m−3 (3.06) and 2.12 mg m−3 (7.74), respectively. One percent of the samples exceeded 15 mg m−3, 6% exceeded 5 mg m−3, and 48% exceeded 1 mg m−3. The number of collected pairs is generally appropriate to detect a 35% difference when outliers (negative mass loadings) are removed. Statistical evaluation of the nonsimilar sampler pair results produced a finding of no significant difference between any pairing of sampler type. A practical consideration for sampling in the USA is that the ACCU-CAP™ is similar to the sampler currently used by the Occupational Safety and Health Administration for purposes of demonstrating compliance with its permissible exposure limit for wood dust, which is the same as for Particles Not Otherwise Regulated, also known as inert dust or nuisance dust (Method PV2121).
doi:10.1093/annhyg/meq075
PMCID: PMC3037778  PMID: 21036895
ACCU-CAP™; Button sampler; CIP10-I sampler; GSP sampler; inhalable sampling; IOM sampler; wood dust
18.  Particle Size Distribution of Airborne Microorganisms and Pathogens during an Intense African Dust Event in the Eastern Mediterranean 
Environmental Health Perspectives  2007;116(3):292-296.
Background
The distribution of microorganisms, and especially pathogens, over airborne particles of different sizes has been ignored to a large extent, but it could have significant implications regarding the dispersion of these microorganisms across the planet, thus affecting human health.
Objectives
We examined the microbial quality of the aerosols over the eastern Mediterranean region during an African storm to determine the size distribution of microorganisms in the air.
Methods
We used a five-stage cascade impactor for bioaerosol collection in a coastal city on the eastern Mediterranean Sea during a north African dust storm. Bacterial communities associated with aerosol particles of six different size ranges were characterized following molecular culture–independent methods, regardless of the cell culturability (analysis of 16S rRNA genes).
Results
All 16S rDNA clone libraries were diverse, including sequences commonly found in soil and marine ecosystems. Spore-forming bacteria such as Firmicutes dominated large particle sizes (> 3.3 μm), whereas clones affiliated with Actinobacteria (found commonly in soil) and Bacteroidetes (widely distributed in the environment) gradually increased their abundance in aerosol particles of reduced size (< 3.3 μm). A large portion of the clones detected at respiratory particle sizes (< 3.3 μm) were phylogenetic neighbors to human pathogens that have been linked to several diseases.
Conclusions
The presence of aerosolized bacteria in small size particles may have significant implications to human health via intercontinental transportation of pathogens.
doi:10.1289/ehp.10684
PMCID: PMC2265054  PMID: 18335093
African dust; bacterial community composition; microbial transport; particle size distribution; pathogens
19.  Methods for Sampling of Airborne Viruses 
Summary: To better understand the underlying mechanisms of aerovirology, accurate sampling of airborne viruses is fundamental. The sampling instruments commonly used in aerobiology have also been used to recover viruses suspended in the air. We reviewed over 100 papers to evaluate the methods currently used for viral aerosol sampling. Differentiating infections caused by direct contact from those caused by airborne dissemination can be a very demanding task given the wide variety of sources of viral aerosols. While epidemiological data can help to determine the source of the contamination, direct data obtained from air samples can provide very useful information for risk assessment purposes. Many types of samplers have been used over the years, including liquid impingers, solid impactors, filters, electrostatic precipitators, and many others. The efficiencies of these samplers depend on a variety of environmental and methodological factors that can affect the integrity of the virus structure. The aerodynamic size distribution of the aerosol also has a direct effect on sampler efficiency. Viral aerosols can be studied under controlled laboratory conditions, using biological or nonbiological tracers and surrogate viruses, which are also discussed in this review. Lastly, general recommendations are made regarding future studies on the sampling of airborne viruses.
doi:10.1128/MMBR.00002-08
PMCID: PMC2546863  PMID: 18772283
20.  Characterization of Airborne Bacteria at an Underground Subway Station 
The reliable detection of airborne biological threat agents depends on several factors, including the performance criteria of the detector and its operational environment. One step in improving the detector's performance is to increase our knowledge of the biological aerosol background in potential operational environments. Subway stations are enclosed public environments, which may be regarded as potential targets for incidents involving biological threat agents. In this study, the airborne bacterial community at a subway station in Norway was characterized (concentration level, diversity, and virulence- and survival-associated properties). In addition, a SASS 3100 high-volume air sampler and a matrix-assisted laser desorption ionization–time of flight mass spectrometry-based isolate screening procedure was used for these studies. The daytime level of airborne bacteria at the station was higher than the nighttime and outdoor levels, and the relative bacterial spore number was higher in outdoor air than at the station. The bacterial content, particle concentration, and size distribution were stable within each environment throughout the study (May to September 2010). The majority of the airborne bacteria belonged to the genera Bacillus, Micrococcus, and Staphylococcus, but a total of 37 different genera were identified in the air. These results suggest that anthropogenic sources are major contributors to airborne bacteria at subway stations and that such airborne communities could harbor virulence- and survival-associated properties of potential relevance for biological detection and surveillance, as well as for public health. Our findings also contribute to the development of realistic testing and evaluation schemes for biological detection/surveillance systems by providing information that can be used to mimic real-life operational airborne environments in controlled aerosol test chambers.
doi:10.1128/AEM.07212-11
PMCID: PMC3298144  PMID: 22247150
21.  Differences in estimates of size distribution of beryllium powder materials using phase contrast microscopy, scanning electron microscopy, and liquid suspension counter techniques 
Accurate characterization of the physicochemical properties of aerosols generated for inhalation toxicology studies is essential for obtaining meaningful results. Great emphasis must also be placed on characterizing particle properties of materials as administered in inhalation studies. Thus, research is needed to identify a suite of techniques capable of characterizing the multiple particle properties (i.e., size, mass, surface area, number) of a material that may influence toxicity. The purpose of this study was to characterize the morphology and investigate the size distribution of a model toxicant, beryllium. Beryllium metal, oxides, and alloy particles were aerodynamically size-separated using an aerosol cyclone, imaged dry using scanning electron microscopy (SEM), then characterized using phase contrast microscopy (PCM), a liquid suspension particle counter (LPC), and computer-controlled SEM (CCSEM). Beryllium metal powder was compact with smaller sub-micrometer size particles attached to the surface of larger particles, whereas the beryllium oxides and alloy particles were clusters of primary particles. As expected, the geometric mean (GM) diameter of metal powder determined using PCM decreased with aerodynamic size, but when suspended in liquid for LPC or CCSEM analysis, the GM diameter decreased by a factor of two (p < 0.001). This observation suggested that the smaller submicrometer size particles attached to the surface of larger particles and/or particle agglomerates detach in liquid, thereby shifting the particle size distribution downward. The GM diameters of the oxide materials were similar regardless of sizing technique, but observed differences were generally significant (p < 0.001). For oxides, aerodynamic cluster size will dictate deposition in the lung, but primary particle size may influence biological activity. The GM diameter of alloy particles determined using PCM became smaller with decreasing aerodynamic size fraction; however, when suspended in liquid for CCSEM and LPC analyses, GM particle size decreased by a factor of two (p < 0.001) suggesting that alloy particles detach in liquid. Detachment of particles in liquid could have significance for the expected versus actual size (and number) distribution of aerosol delivered to an exposure subject. Thus, a suite of complimentary analytical techniques may be necessary for estimating size distribution. Consideration should be given to thoroughly understanding the influence of any liquid vehicle which may alter the expected aerosol size distribution.
doi:10.1186/1743-8977-4-3
PMCID: PMC1810321  PMID: 17328812
22.  The Abbreviated Impactor Measurement (AIM) Concept: Part 1—Influence of Particle Bounce and Re-Entrainment—Evaluation with a “Dry” Pressurized Metered Dose Inhaler (pMDI)-Based Formulation 
AAPS PharmSciTech  2009;10(1):243-251.
The abbreviated impactor measurement concept is a potential improvement to the labor-intensive full-resolution cascade impactor methodology for inhaler aerosol aerodynamic particle size distribution (APSD) measurement by virtue of being simpler and therefore quicker to execute. At the same time, improved measurement precision should be possible by eliminating stages upon which little or no drug mass is collected. Although several designs of abbreviated impactor systems have been developed in recent years, experimental work is lacking to validate the technique with aerosols produced by currently available inhalers. In part 1 of this two-part article that focuses on aerosols produced by pressurized metered dose inhalers (pMDIs), the evaluation of two abbreviated impactor systems (Copley fast screening Andersen impactor and Trudell fast screening Andersen impactor), based on the full-resolution eight-stage Andersen nonviable cascade impactor (ACI) operating principle, is reported with a formulation producing dry particles. The purpose was to investigate the potential for non-ideal collection behavior associated with particle bounce in relation to internal losses to surfaces from which particles containing active pharmaceutical ingredient are not normally recovered. Both abbreviated impactors were found to be substantially equivalent to the full-resolution ACI in terms of extra-fine and fine particle and coarse mass fractions used as metrics to characterize the APSD of these pMDI-produced aerosols when sampled at 28.3 L/min, provided that precautions are taken to coat collection plates to minimize bounce and entrainment.
doi:10.1208/s12249-009-9202-9
PMCID: PMC2663692  PMID: 19280348
cascade impactor; inhaler testing; particle bounce; particle re-entrainment; particle size distribution
23.  Size-resolved emission rates of airborne bacteria and fungi in an occupied classroom 
Indoor Air  2012;22(4):339-351.
The role of human occupancy as a source of indoor biological aerosols is poorly understood. Size-resolved concentrations of total and biological particles in indoor air were quantified in a classroom under occupied and vacant conditions. Per-occupant emission rates were estimated through a mass-balance modeling approach, and the microbial diversity of indoor and outdoor air during occupancy was determined via rDNA gene sequence analysis. Significant increases of total particle mass and bacterial genome concentrations were observed during the occupied period compared to the vacant case. These increases varied in magnitude with the particle size and ranged from 3 to 68 times for total mass, 12–2700 times for bacterial genomes, and 1.5–5.2 times for fungal genomes. Emission rates per person-hour because of occupancy were 31 mg, 37 × 106 genome copies, and 7.3 × 106 genome copies for total particle mass, bacteria, and fungi, respectively. Of the bacterial emissions, ∼18% are from taxa that are closely associated with the human skin microbiome. This analysis provides size-resolved, per person-hour emission rates for these biological particles and illustrates the extent to which being in an occupied room results in exposure to bacteria that are associated with previous or current human occupants.
Practical Implications
Presented here are the first size-resolved, per person emission rate estimates of bacterial and fungal genomes for a common occupied indoor space. The marked differences observed between total particle and bacterial size distributions suggest that size-dependent aerosol models that use total particles as a surrogate for microbial particles incorrectly assess the fate of and human exposure to airborne bacteria. The strong signal of human microbiota in airborne particulate matter in an occupied setting demonstrates that the aerosol route can be a source of exposure to microorganisms emitted from the skin, hair, nostrils, and mouths of other occupants.
doi:10.1111/j.1600-0668.2012.00769.x
PMCID: PMC3437488  PMID: 22257156
Bioaerosols; Resuspension; qPCR; Particle size distribution; Indoor microbiome; 454 pyrosequencing; Human microbiome
24.  Experimental determination of the respiratory tract deposition of diesel combustion particles in patients with chronic obstructive pulmonary disease 
Background
Air pollution, mainly from combustion, is one of the leading global health risk factors. A susceptible group is the more than 200 million people worldwide suffering from chronic obstructive pulmonary disease (COPD). There are few data on lung deposition of airborne particles in patients with COPD and none for combustion particles.
Objectives
To determine respiratory tract deposition of diesel combustion particles in patients with COPD during spontaneous breathing.
Methods
Ten COPD patients and seven healthy subjects inhaled diesel exhaust particles generated during idling and transient driving in an exposure chamber. The respiratory tract deposition of the particles was measured in the size range 10–500 nm during spontaneous breathing.
Results
The deposited dose rate increased with increasing severity of the disease. However, the deposition probability of the ultrafine combustion particles (< 100 nm) was decreased in COPD patients. The deposition probability was associated with both breathing parameters and lung function, but could be predicted only based on lung function.
Conclusions
The higher deposited dose rate of inhaled air pollution particles in COPD patients may be one of the factors contributing to their increased vulnerability. The strong correlations between lung function and particle deposition, especially in the size range of 20–30 nm, suggest that altered particle deposition could be used as an indicator respiratory disease.
doi:10.1186/1743-8977-9-30
PMCID: PMC3464711  PMID: 22839109
Lung deposition; Toxicity; Health effects; Air pollution; Agglomerate; Nanoparticles; Aerosol; COPD; Diesel exhaust
25.  Direct Deposition of Gas Phase Generated Aerosol Gold Nanoparticles into Biological Fluids - Corona Formation and Particle Size Shifts 
PLoS ONE  2013;8(9):e74702.
An ongoing discussion whether traditional toxicological methods are sufficient to evaluate the risks associated with nanoparticle inhalation has led to the emergence of Air-Liquid interface toxicology. As a step in this process, this study explores the evolution of particle characteristics as they move from the airborne state into physiological solution. Airborne gold nanoparticles (AuNP) are generated using an evaporation-condensation technique. Spherical and agglomerate AuNPs are deposited into physiological solutions of increasing biological complexity. The AuNP size is characterized in air as mobility diameter and in liquid as hydrodynamic diameter. AuNP:Protein aggregation in physiological solutions is determined using dynamic light scattering, particle tracking analysis, and UV absorption spectroscopy. AuNPs deposited into homocysteine buffer form large gold-aggregates. Spherical AuNPs deposited in solutions of albumin were trapped at the Air-Liquid interface but was readily suspended in the solutions with a size close to that of the airborne particles, indicating that AuNP:Protein complex formation is promoted. Deposition into serum and lung fluid resulted in larger complexes, reflecting the formation of a more complex protein corona. UV absorption spectroscopy indicated no further aggregation of the AuNPs after deposition in solution. The corona of the deposited AuNPs shows differences compared to AuNPs generated in suspension. Deposition of AuNPs from the aerosol phase into biological fluids offers a method to study the protein corona formed, upon inhalation and deposition in the lungs in a more realistic way compared to particle liquid suspensions. This is important since the protein corona together with key particle properties (e.g. size, shape and surface reactivity) to a large extent may determine the nanoparticle effects and possible translocation to other organs.
doi:10.1371/journal.pone.0074702
PMCID: PMC3785473  PMID: 24086363

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