Fungal spore source strength tester: laboratory evaluation of a new concept

The airborne fungal spore concentration measured with air samplers during specific time intervals does not always adequately represent the maximum spore concentration levels, because of the sporadic nature of spore release. Hence, a reliable method is needed to directly assess the indoor fungal sources with respect to their spore aerosolization potential. In this study, the newly developed fungal spore source strength tester (FSSST), which aerosolizes spores from growth surfaces and samples the airborne fungi into a bioaerosol sampler, was evaluated in the laboratory. The FSSST's operational flow rates of 30 and 12.5 l/min were tested. The fungal spores released from moldy surfaces were measured with an optical particle counter. Simultaneously, the spores were collected by a bioaerosol sampler: either with a 37-mm filter cassette or with the BioSampler. Three material types, ceiling tile, gypsum board and plastic sheet coated with agar, were tested after they were inoculated with the fungus Aspergillus versicolor. In addition, gypsum board naturally contaminated with various fungi (obtained from a mold-problem home) was tested in the laboratory using the FSSST. In all three laboratory-inoculated materials, the release rate of A. versicolor was found to be higher when the FSSST operated at 30 l/min than at 12.5 l/min. Nevertheless, even at 12.5 l/min the number of spores aerosolized from the source during 10 min was found sufficient to reflect the highest level of release that may occur in indoor environments. At 12.5 l/min, the release rate of A. versicolor during the first 10-min period was (23.9 +/- 17.7)x10(4) cm(-2) for ceiling tile, (1.3 +/- 0.3)x10(4) cm(-2) for gypsum board and (0.13 +/- 0.08)x10(4) cm(-2) for agar surface (based on the samples collected with the BioSampler). The spore release rate was higher during the first 10 min than during the second 10 min of the FSSST application. It was observed that the particles aerosolized from the A. versicolor culture included spore aggregates and single spores, as well as mycelial fragments. Overall, 0.6 +/- 0.3% of spores detected on 1 cm2 of ceiling tile inoculated with A. versicolor were aerosolized during the 10-min source testing. The respective number was 9.2 +/- 1.0% for the laboratory-inoculated gypsum board, 0.002 +/- 0.001% for the laboratory-inoculated plastic covered with agar and 1.8 +/- 0.2% for naturally contaminated gypsum board. Our data suggest that the FSSST provides very favorable conditions for the spore aerosolization and thus can be used in the field to assess the maximum potential spore release from a fungal source.     

Exposure matrices of endotoxin, (1 → 3)-β-d-glucan, fungi, and dust mite allergens in flood-affected homes of New Orleans

This study examined: (i) biocontaminant levels in flooded homes of New Orleans two years after the flooding; (ii) seasonal changes in biocontaminant levels, and (iii) correlations between biocontaminant levels obtained by different environmental monitoring methods. Endotoxin, (1 → 3)-β-d-glucan, fungal spores, and dust mite allergens were measured in 35 homes during summer and winter. A combination of dust sampling, aerosolization-based microbial source assessment, and long-term inhalable bioaerosol sampling aided in understanding exposure matrices. On average, endotoxin found in the aerosolized fraction accounted for < 2% of that measured in the floor dust, suggesting that vacuuming could overestimate inhalation exposures. In contrast, the (1 → 3)-β-d-glucan levels in the floor dust and aerosolized fractions were mostly comparable, and 25% of the homes showed aerosolizable levels even higher than the dust-borne levels. The seasonal patterns for endotoxin in dust and the aerosolizable fraction were different from those found for (1 → 3)-β-d-glucan, reflecting the temperature and humidity effects on bacterial and fungal contamination. While the concentration of airborne endotoxin followed the same seasonal trend as endotoxin aerosolized from surfaces, no significant seasonal difference was identified for the concentrations of airborne (1 → 3)-β-d-glucan and fungal spores. This was attributed to the difference in the particle size; smaller endotoxin-containing particles can remain airborne for longer time than larger fungal spores or (1 → 3)-β-d-glucan-containing particles. It is also possible that fungal aerosolization in home environments did not reach its full potential. Detectable dust mite allergens were found only in dust samples, and more commonly in occupied homes. Levels of endotoxin, (1 → 3)-β-d-glucan, and fungi in air had decreased during the two-year period following the flooding as compared to immediate measurements; however, the dust-borne endotoxin and (1 → 3)-β-d-glucan levels remained elevated. No conclusive correlations were found between the three environmental monitoring methods. The findings support the use of multiple methods when assessing exposure to microbial contaminants.     

Relationship between indoor and outdoor bio-aerosols collected with a button inhalable aerosol sampler in urban homes

This field study investigated the relationship between indoor and outdoor concentrations of airborne actinomycetes, fungal spores, and pollen. Air samples were collected for 24 h with a button inhalable aerosol sampler inside and outside of six single-family homes located in the Cincinnati area (overall, 15 pairs of samples were taken in each home). The measurements were conducted during three seasons - spring and fall 2004, and winter 2005. The concentration of culturable actinomycetes was mostly below the detection limit. The median indoor/outdoor ratio (I/O) for actinomycetes was the highest: 2.857. The indoor of fungal and pollen concentrations followed the outdoor concentrations while indoor levels were mostly lower than the outdoor ones. The I/O ratio of total fungal spores (median=0.345) in six homes was greater than that of pollen grains (median=0.025). The low I/O ratios obtained for pollen during the peak ambient pollination season (spring) suggest that only a small fraction penetrated from outdoor to indoor environment. This is attributed to the larger size of pollen grains. Higher indoor concentration levels and variability in the I/O ratio observed for airborne fungi may be associated with indoor sources and/or higher outdoor-to-indoor penetration of fungal spores compared to pollen grains. Practical Implication This study addresses the relationship between indoor and outdoor concentrations of three different types of bio-aerosols, namely actinomycetes, fungal spores, and pollen grains. The results show that actinomycetes are rare in indoor and outdoor air in Midwest, USA. Exposure to pollen occurs mainly in the outdoor air even during peak pollen season. Unexpectedly high fungal spore concentrations were measured outdoors during winter. The presented pilot database on the inhalable levels of indoor and outdoor bio-aerosols can help apportion and better characterize the inhalation exposure to these bio-aerosols. Furthermore, the data can be incorporated into existing models to quantify the penetration of biological particles into indoor environments from outdoors.     

Association between indoor mold and asthma among children in Buffalo, New York

Asthma is a leading chronic disease among children and places a significant burden on public health. Exposure to indoor mold has been associated with asthma symptoms. However, many mold assessments have relied on visual or other identification of damp conditions and mold presence, thus have not examined associations with specific fungal genera. The objective of this case-control study was to examine the relationship between airborne mold concentrations and asthma status among children and to identify the contribution from specific mold genera in air. Participants completed a questionnaire of home environmental conditions and underwent indoor air sampling in the home, from which viable and total-count fungal spores were quantified. The most prevalent fungi in the homes were the allergenic molds Cladosporium (98% and 87% of homes from viable and total count samples, respectively) and Penicillium (91% and 73%). There were no significant differences in mean fungal concentrations between the homes of cases and controls, although the observed rate of exposure to several molds was higher among the cases. Among children who lacked a family history of asthma, cases had significantly higher exposures to viable Aspergillus. Measured humidity levels in the home corresponded with some self-reported indicators of mold and dampness. PRACTICAL IMPLICATIONS: The results of this study support existing literature that indoor fungal exposures play a role in current asthma status and that some qualitative assessments of mold exposure correspond to fungi present in indoor air.     

The effects of meteorological factors on atmospheric bioaerosol concentrations--a review

Over land surfaces a quarter of the total airborne particulate may be made up of biological material in the form of pollen, fungal spores, bacteria, viruses, or fragments of plant and animal matter. Meteorological variables affect the initial release of this material and its dispersal once airborne. Temperature and water availability will affect the size of the source and will control the release of some actively released fungal spores. Inertly released material will become airborne when the drying of the surface reduces bonding forces, and when the material is disrupted by sufficiently strong air movement or by mechanical disturbance. The wind speed necessary to disrupt material is noted to be less on a plant surface than on the ground surface. Measurements of the concentrations of airborne material near dominant sources are reviewed for both area sources, and for point sources such as sewage and waste treatment works, agricultural practices, and diseased animals. The concentration of airborne material remote from sources is considered along with the effects of on and off shore winds and some examples of long distance transport of material. The vertical concentration of bacteria is noted to decline less rapidly than that of fungal spores. The short-term variation of pollen, fungal spore, and bacterial concentrations are also considered.     

A pilot study to investigate the effects of combined dehumidification and HEPA filtration on dew point and airborne mold spore counts in day care centers

Meteorological factors such as relative humidity directly correlate with airborne fungal levels outdoors and indoors. While dehumidification alone is effective at reducing moisture necessary for mold growth, it is inadequate as a single intervention as it does not remove viable and non-viable fungal spores that are potentially allergenic. The purpose of this pilot study was to investigate whether dehumidification in combination with high-efficiency particulate arrestance (HEPA) filtration is effective at reducing airborne mold spore levels in day care centers. Two day care centers within a 2-mile radius of each other were selected. Day care center A was 2 years old with eight rooms while day care center B was 15 years old with six rooms. A high efficiency Santa Fe dehumidification unit equipped with a HEPA filter was installed in half the rooms (intervention) of each day care facility. Electronic HOBO data loggers continuously measured outdoor and indoor room dew point and temperature every 2 h throughout the study. Dew point and airborne fungal spore measurements from selected rooms with controlled air conditions were analyzed by comparing baseline measurements to those obtained at subsequent time periods over 1 year. Regression models accounted for correlations between measurements in the same room over time. Intervention resulted in a lowered average dew point from baseline by 8.8 degrees C compared with a decrease of 7.1 degrees C in non-intervention rooms across all time periods in both facilities (P<0.001). Fungal analyses demonstrated lower baseline (P=0.06) and follow-up means in intervention rooms (P<0.05), however the change from baseline to end of follow-up differed between intervention and non-intervention rooms in the two facilities. Log transformation was applied to approximate normality of fungal measurements. Dehumidification with HEPA filtration was effective at controlling indoor dew point in both facilities and at reducing airborne culturable fungal spore levels in one of the two facilities. These preliminary results provide a scientific rationale for using this intervention in future studies designed to investigate the impact of indoor mold exposure on health outcomes. PRACTICAL IMPLICATIONS: Poor indoor air quality is a recognized cause or contributing factor to health effects. Dampness and humidity have been linked to upper and lower respiratory symptoms in children and adults. This study indicates that reducing indoor relative humidity and airborne mold spore levels using high-efficiency dehumidification units equipped with HEPA filtration is feasible even in work facilities such as day care centers where traffic in and out of the building is difficult to regulate. Clinicians should emphasize to their patients the importance of dehumidification and HEPA filtration to improve indoor air quality in the home and workplace.     

Measuring detachment of Aspergillus niger spores from colonies with an atomic force microscope

Detachment of fungal spores from moldy surfaces and the subsequent aerosolization can lead to adverse health effects. Spore aerosolization occurs when the forces for aerosolization exceed the binding forces of spores with their colonies. The threshold force to detach a spore from a growing colony remains unknown. This investigation measured the detachment of spores of Aspergillus niger from a colony using an atomic force microscope (AFM). The spores were first affixed to the cantilever of the AFM with ultraviolet curing glue, and then, the colony was moved downward until the spores detached. The threshold detachment forces were inferred from the deflection of the cantilever. In addition, the spores were aerosolized in a wind tunnel by a gradual increase of the blowing air speed. The forces measured by the AFM were compared with the hydrodynamic forces for aerosolization. The AFM measurements revealed that a force of 3.27 ± 0.25 nN was required to detach a single spore from the 4‐day‐old colony, while 1.98 ± 0.13 nN was sufficient for the 10‐day‐old colony. Slightly smaller detachment forces were observed by the AFM than were determined by the aerosolization tests.     

Microbial contamination of indoor air due to leakages from crawl space: a field study

Mechanical exhaust ventilation system is typical in apartment buildings in Finland. In most buildings the base floor between the first floor apartments and crawl space is not air tight. As the apartments have lower pressure than the crawl space due to ventilation, contaminated air may flow from the crawl space to the apartments. The object of this study was to find out whether a potential air flow from crawl space has an influence on the indoor air quality. The results show that in most cases the concentration of fungal spores was clearly higher in the crawl space than inside the building. The size distribution of fungal spores depended on the fungal species. Correlation between the fungal spores in the crawl space and indoors varied with microbial species. Some species have sources inside the building, which confounds the possible relation between crawl pace and indoor concentrations. Some species, such as Acremonium, do not normally have a source indoors, but its concentration in the crawl space was elevated; our measurements showed also elevated concentrations of Acremonium in the air of the apartments. This consistent finding shows a clear linkage between fungal spores in the indoor air and crawl space. We conclude that a building with a crawl space and pressure difference over the base floor could be a potential risk for indoor air quality in the first floor apartments.     

Determination of fungal spore release from wet building materials

The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species of typical indoor fungi has been measured under controlled conditions. The fungi were cultivated for a period of 4-6 weeks on sterilized wet wallpapered gypsum boards at a relative humidity (RH) of approximately 97%. A specially designed small chamber (P-FLEC) was placed on the gypsum board. The release of fungal spores was induced by well-defined jets of air impacting from rotating nozzles. The spores and other particles released from the surface were transported by the air flowing from the chamber through a top outlet to a particle counter and sizer. For two of the fungi (Penicillium chrysogenum and Trichoderma harzianum), the number of spores produced on the gypsum board and subsequently released was quantified. Also the relationship between air velocities from 0.3 to 3 m/s over the surface and spore release has been measured. The method was found to give very reproducible results for each fungal isolate, whereas the spore release is very different for different fungi under identical conditions. Also, the relationship between air velocity and spore release depends on the fungus. For some fungi a significant number of particles smaller than the spore size were released. The method applied in the study may also be useful for field studies and for generation of spores for exposure studies.     

A comparison of airborne ergosterol, glucan and Air-O-Cell data in relation to physical assessments of mold damage and some other parameters

We report here a comparison of long duration air samples in 110 homes where the material collected on open faced filter cassettes was analyzed for beta 1,3-d glucan, ergosterol, cholesterol and endotoxin. These data were then compared to careful estimates of visible mold and Air-O-Cell data. All the values found except cholesterol were of a similar magnitude to values reported in the limited number of studies available. Glucan was measured with a factor G based assay of the Limulus Amebocyte Lysate followed by size exclusion chromatography. This showed that the majority of airborne glucan found in these houses was fungal in origin arising from both yeasts and intact spores, as well as hyphal and spore fragments. Hyphal and spore fragments together represented 16% of the intact spore counts but over a broad range. Correlations between airborne glucan were strong for ergosterol and visible mold. However, airborne ergosterol was more highly correlated to visible mold than glucan. Endotoxin and Air-O-Cell measurements were poorly or not related to the other measures in the study. This study provides confidence that long duration air samples of the toxin glucan and ergosterol are related to building damage. PRACTICAL IMPLICATIONS: Some studies of damp buildings have shown a relationship between extent of water/mold damage and symptoms. This study compared long duration air samples for glucan and ergosterol to extent of visible mold in houses measuring also the nature of the glucans present. Both measures were highly correlated to extent of visible mold damage in the houses; ergosterol was somewhat superior. Spore counts or prevalence of Asp/Pen in Air-O-Cell samples was not related to extent of visible mold damage but the observation of hyphal fragments was more likely when mold damage was present. This indicates that rigorous assessment of mold damage is a useful measure.     

Use of a robotic sampling platform to assess young children's exposure to indoor bioaerosols

Indoor exposures to allergens, mold spores, and endotoxin have been suggested as etiological agents of asthma; therefore, accurate determination of those exposures, especially in young children (6-36 months), is important for understanding the development of asthma. Because use of personal sampling equipment in this population is difficult, and in children <1 year of age impossible, we developed a personal sampling surrogate: the Pretoddler Inhalable Particulate Environmental Robotic (PIPER) sampler to better estimate their exposures. During sampling, PIPER simulates the activity patterns, speed of motion, and the height of the breathing zones of young children, and mechanically resuspends the deposited dust just as a young child does during running and crawling. The concentrations of allergens, mold spores, and endotoxin measured by PIPER were compared to those measured using traditional stationary air sampling method in 75 homes in central New Jersey, United States. Endotoxin was detected in all homes with median concentrations of 1.0 and 0.55 EU/m(3) for PIPER and stationary sampler, respectively. The difference in median concentrations obtained using the two methods was statistically significant for homes with carpeted floors (P = 0.0001) in the heating season. For such homes, the average ratio of endotoxin concentration measured by PIPER to the stationary sampler was 2.96 (95% CI 2.29-3.63). Fungal spores were detected in all homes, with median fungal concentrations of 316 and 380 spores/m(3) for PIPER and stationary sampler, respectively. For fungi, the difference between the two sampling methods was not statistically significant. For both sampling methods, the total airborne mold levels were statistically significantly higher in the non-heating season than in the heating season. Allergens were detected in ~15% of investigated homes. The data indicate that the traditional stationary air-sampling methods may substantially underestimate personal exposures to endotoxin, especially due to resuspension of dust from carpeted floor surfaces. A personal sampling surrogate, such as PIPER, is a feasible approach to estimate personal exposures in young children. PIPER should be seriously considered as the sampling platform for future exposure studies in young children. PRACTICAL IMPLICATIONS: This study investigated potential indoor bioaerosol exposure of young children using a Pretoddler Inhalable Particulate Environmental Robotic (PIPER) sampler platform. The results show that the traditional stationary air-sampling methods can substantially underestimate personal exposures to resuspended material, and that a personal sampling surrogate, such as PIPER, offers a feasible surrogate for measuring personal inhalation exposures of young children.     

Home Humidifiers as a Potential Source of Exposure to Microbial Pathogens,Endotoxins, and Allergens

The propensity of various types of home humidifiers to support and disseminate microbial contaminants into indoor air was tested. Reservoir water and air discharged from humidifiers seeded in the laboratory or naturally contaminated in the home were analyzed by standard microbiological methods. Clinically insignificant as well as overt or potentially pathogenic microorganisms were found to colonize the reservoirs of all types of humidifiers, but only cool mist and ultrasonic units readily aerosolized bacteria and endotoxin. Only cool mist units emitted hydrophobic fungal spores. Cool mist units discharged the greatest number of water particles in the inhalabk size range (4–16 μm) while ultrasonic units were more likely to emit respirable-sized water particles (< 0.2–4 μm). Overt pathogens isolated from humidifiers in homes included Legionella and a pathogenic Acanthamoeba. Aerosolizing humidifiers should thus be avoided if frequent, thorough cleaning of the units is not practical.     

Changes in profiles of airborne fungi in flooded homes in southern Taiwan after Typhoon Morakot

In August 2009, the historic Typhoon Morakot brought extreme rainfall and resulted in flooding which spread throughout southern Taiwan. This study compared the difference between fungal concentrations before and after the disaster in selected homes of the Tainan metropolitan area, which were hit hardest by the catastrophe. A group of 83 households available from a prior cohort established with random sampling out of a regional population in southern Taiwan was contacted successfully by telephone. Twenty-five of these reported to have suffered from floods of various degrees at this time. Around 2 weeks after the event, at which time most of the remedial process had been completed by self-efforts and public health endeavours, 14 of these 25 (56%) agreed to participate in measurements of the airborne microbial concentrations. The averages (standard deviation) of the total culturable fungal concentrations in children's bedrooms and flooded rooms were 18,181 (25,854) colony-forming units per cubic metre (CFU/m³) and 13,440 (11,033) CFU/m³, respectively. The airborne fungal spore levels in the 2 above-mentioned indoor sites were 221,536 (169,640) spores/m³ and 201,582 (137,091) spores/m³, respectively. The average indoor/outdoor ratios in the children's bedrooms were 4.2 for culturable fungi and 1.4 for fungal spores. These values were higher than the respective values measured in the same homes during the previous year: 1.1 and 0.6. In terms of the specific fungal profile, the percentages of Aspergillus spp. increased significantly in both the indoor and outdoor environments after the event. To this date, this study is among the limited research that has been conducted to quantitatively demonstrate that fungal manifestation is likely to persist in flooded homes even after seemingly robust remedial measures have been put into place. Studies to examine the potential health implications and effectiveness of better remedial technology remain much needed.     

Airborne viable, non-viable, and allergenic fungi in a rural agricultural area of India: a 2-year study at five outdoor sampling stations

The information on airborne allergenic fungal flora in rural agricultural areas is largely lacking. Adequate information is not available to the bioaerosol researchers regarding the choice of single versus multiple sampling stations for the monitoring of both viable and non-viable airborne fungi. There is no long-term study estimating the ratios of viable and non-viable fungi in the air and earlier studies did not focus on the fractions of airborne allergenic fungi with respect to the total airborne fungal load. To fill these knowledge gaps, volumetric paired assessments of airborne viable and non-viable fungi were performed in five outdoor sampling stations during two consecutive years in a rural agricultural area of India. Samples were collected at 10-day intervals by the Burkard Personal Slide Sampler and the Andersen Two-Stage Viable Sampler. The data on the concentrations of total and individual fungal types from five stations and 2 different years were analyzed and compared by statistical methods. The allergenicity of the prevalent airborne viable fungi was estimated by the skin-prick tests of >100 rural allergy patients using the antigenic fungal extracts from isolates collected with the Andersen sampler. The ranges of total fungal spore concentration were 82-2365 spores per cubic meter of air (spores/m3) in the first sampling year and 156-2022 spores/m3 in the second sampling year. The concentration ranges of viable fungi were 72-1796 colony-forming units per cubic meter of air (CFU/m3) in the first sampling year and 155-1256 CFU/m3 in the second sampling year. No statistically significant difference was observed between the total spore data of the 2 years, however, the data between five stations showed a significant difference (P<0.0001). No statistically significant difference existed between stations and years with respect to the concentration of viable fungi. When the data of individual allergenic fungal concentrations were compared between stations and years, no statistically significant difference was observed in all cases except for Aspergillus japonicus and Rhizopus nigricans, which showed significant difference in case of stations and years, respectively. The ratios between the total fungal spores collected by the Burkard sampler and the viable fungi collected by the Andersen sampler from all sampling stations ranged between 0.29 and 7.61. The antigenic extracts of eight prevalent viable airborne fungi (A. flavus, A. japonicus, A. fumigatus, Alternaria alternata, Cladosporium cladosporioides, Curvularia pallescens, Fusarium roseum, and R. nigricans) demonstrated >60% positive reactions in the skin prick test. These selected allergenic fungi collectively represented 31.7-63.2% of the total airborne viable fungi in different stations. The study concluded that: (i) a rich fungal airspora existed in the rural study area, (ii) to achieve representative information on the total airborne fungal spores of an area, the monitoring in multiple sampling stations is preferable over a single sampling station; for viable fungi, however, one station can be considered, (iii) the percentage of airborne fungal viability is higher in rural agricultural areas, and (iv) approximately 52% of the viable airborne fungi in the rural study area were allergenic.     

Characterization and pro-inflammatory potential of indoor mold particles

A number of epidemiological studies find an association between indoor air dampness and respiratory health effects. This is often suggested to be linked to enhanced mold growth. However, the role of mold is obviously difficult to disentangle from other dampness-related exposure including microbes as well as non-biological particles and chemical pollutants. The association may partly be due to visible mycelial growth and a characteristic musty smell of mold. Thus, the potential role of mold exposure should be further explored by evaluating information from experimental studies elucidating possible mechanistic links. Such studies show that exposure to spores and hyphal fragments may act as allergens and pro-inflammatory mediators and that they may damage airways by the production of toxins, enzymes, and volatile organic compounds. In the present review, we hypothesize that continuous exposure to mold particles may result in chronic low-grade pro-inflammatory responses contributing to respiratory diseases. We summarize some of the main methods for detection and characterization of fungal aerosols and highlight in vitro research elucidating how molds may induce toxicity and pro-inflammatory reactions in human cell models relevant for airway exposure. Data suggest that the fraction of fungal hyphal fragments in indoor air is much higher than that of airborne spores, and the hyphal fragments often have a higher pro-inflammatory potential. Thus, hyphal fragments of prevalent mold species with strong pro-inflammatory potential may be particularly relevant candidates for respiratory diseases associated with damp/mold-contaminated indoor air. Future studies linking of indoor air dampness with health effects should assess the toxicity and pro-inflammatory potential of indoor air particulate matter and combined this information with a better characterization of biological components including hyphal fragments from both pathogenic and non-pathogenic mold species. Such studies may increase our understanding of the potential role of mold exposure.     

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.     

Fungal aerosol composition in moldy basements

Experimental aerosolization studies revealed that fungal fragments including small fragments in the submicrometer size are released from fungal cultures and have been suggested to represent an important fraction of overall fungal aerosols in indoor environments. However, their prevalence indoors and outdoors remains poorly characterized. Moldy basements were investigated for airborne fungal particles including spores, submicron fragments, and larger fragments. Particles were collected onto poly‐L‐lysine‐coated polycarbonate filters and qualitatively and quantitatively analyzed using immunogold labeling combined with field emission scanning electron microscopy. We found that the total fungal aerosol levels including spores, submicrometer, and larger fragments in the moldy basements (median: 80 × 10³ m^?3) were not different from that estimated in control basements (63 × 10³ m^?3) and outdoor (90 × 10³ m^?3). However, mixed effect modeling of the fungal aerosol composition revealed that the fraction of fragments increased significantly in moldy basements, versus the spore fraction that increased significantly in outdoor air. These findings provide new insight on the compositional variation of mixed fungal aerosols in indoor as compared to outdoor air. Our results also suggest that further studies, aiming to investigate the role of fungal aerosols in the fungal exposure‐disease relationships, should consider the mixed composition of various types of fungal particles.     

Collection efficiencies of an electrostatic sampler with superhydrophobic surface for fungal bioaerosols

We recently developed an electrostatic precipitator with superhydrophobic surface (EPSS), which collects particles into a 10- to 40-μl water droplet allowing achievement of very high concentration rates (defined as the ratio of particle concentration in the collection liquid vs. the airborne particle concentration per time unit) when sampling airborne bacteria. Here, we analyzed the performance of this sampler when collecting three commonly found fungal spores--Cladosporium cladosporioides, Penicillium melinii, and Aspergillus versicolor--under different operating conditions. We also adapted adenosine triphosphate (ATP)-based bioluminescence for the analysis of collection efficiency and the concentration rates. The collection efficiency ranged from 10 to 36% at a sampling flow rate of 10 l/min when the airborne fungal spore concentration was approximately 10(5)-10(6) spores/m(3) resulting in concentration rates in the range of 1 × 10(5)-3 × 10(5)/min for a 10-μl droplet. The collection efficiency was inversely proportional to the airborne spore concentration and it increased to above 60% for common ambient spore concentrations, e.g., 10(4)-10(5) spores/m(3). The spore concentrations determined by the ATP-based method were not statistically different from those determined by microscopy and allowed us to analyze spore concentrations that were too low to be reliably detected by microscopy. PRACTICAL IMPLICATIONS: The new electrostatic precipitator with superhydrophobic surface (EPSS) collects airborne fungal spores into small water droplets (10 and 40 μl) allowing achievement of concentration rates that are higher than those of most currently available bioaerosol samplers. Biosamplers with high concentration rates enable detection of low ambient aerial bioaerosol concentrations in various environments, including indoors air, and would be useful for improved exposure assessment. A successful adaptation of the adenosine triphosphate (ATP)-based bioluminescence assay for the quantification of fungal spores from a specific species enables fast sample analysis in laboratory investigations. This rapid assay could be especially useful when investigating the performance of biological samplers as a function of multiple operational parameters.     

Isolation of Aspergillus fumigatus from sputum is associated with elevated airborne levels in homes of patients with asthma

Indoor bioaerosols, such as mold spores, have been associated with respiratory symptoms in patients with asthma; however, dose–response relationships and guidelines on acceptable levels are lacking. Furthermore, a causal link between mold exposure and respiratory infections or asthma remains to be established. The aim of this study was to determine indoor concentrations of Aspergillus fumigatus and a subset of clinically relevant fungi in homes of people with asthma, in relation to markers of airways colonization and sensitization. Air and dust samples were collected from the living room of 58 properties. Fungal concentrations were quantified using mold‐specific quantitative PCR and compared with traditional microscopic analysis of air samples. Isolation of A. fumigatus from sputum was associated with higher airborne concentrations of the fungus in patient homes (P = 0.04), and a similar trend was shown with Aspergillus/Penicillium‐type concentrations analyzed by microscopy (P = 0.058). No association was found between airborne levels of A. fumigatus and sensitization to this fungus, or dustborne levels of A. fumigatus and either isolation from sputum or sensitization. The results of this study suggest that the home environment should be considered as a potential source of fungal exposure, and elevated home levels may predispose people with asthma to airways colonization.