Assessment of indoor bioaerosol exposure using direct-reading versus traditional methods-potential application to home health care.

Home healthcare workers (HHCWs) can be occupationally exposed to bioaerosols in their clients' homes. However, choosing the appropriate method to measure bioaerosol exposures remains a challenge. Therefore, a systematic comparison of existing measurement approaches is essential. Bioaerosol measurements with a real-time, fluorescence-based Wideband Integrated Bioaerosol Sensor (WIBS) were compared to measurements with four traditional off-line methods (TOLMs). The TOLMS included optical microscopic counting of spore trap samples, microbial cultivation of impactor samples, qPCR, and next-generation sequencing (NGS) of filter samples. Measurements were conducted in an occupied apartment simulating the environments that HHCWs could encounter in their patients' homes. Descriptive statistics and Spearman's correlation test were computed to compare the real-time measurement with those of each TOLM. The results showed that the geometric mean number concentrations of the total fluorescent aerosol particles (TFAPs) detected with the WIBS were several orders of magnitude higher than those of total fungi or bacteria measured with the TOLMs. Among the TOLMs, concentrations obtained with qPCR and NGS were the closest to the WIBS detections. Correlations between the results obtained with the WIBS and TOLMs were not consistent. No correlation was found between the concentrations of fungi detected using microscopic counting and any of the WIBS fluorescent aerosol particle (FAP) types, either indoors or outdoors. In contrast, the total concentrations detected with microbial cultivation correlated with the WIBS TFAP results, both indoors and outdoors. Outdoors, the total concentration of culturable bacteria correlated with FAP-type AC. In addition, fungal and bacterial concentrations obtained with qPCR correlated with FAP types AB and AC. For a continuous, high-time resolution but broad scope, the real-time WIBS could be considered, whereas a TOLM would be the best choice for specific and more accurate microbial characterization. HHCWs' activities tend to re-aerosolize bioaerosols causing wide temporal variation in bioparticle concentrations. Thus, the advantage of using the real-time instrument is to capture those variations. This study lays a foundation for future exposure assessment studies targeting HHCWs.

The mycobiomes and bacteriomes of sputum, saliva, and home dust.

Respiratory microbiome is an understudied area of research compared to other microbiomes of the human body. The respiratory tract is exposed to an array of environmental pollutants, including microbes. Yet, we know very little about the relationship between environmental and respiratory microbiome. The primary aim of our study was to compare the mycobiomes and bacteriomes between three sample types from the same participants, including home dust, saliva, and sputum. Samples were collected from 40 adolescents in a longitudinal cohort. We analyzed the samples using 16s bacterial rDNA and ITS fungal rDNA gene sequencing, as well as quantitative PCR with universal fungal and bacterial primers. Results showed that home dust had the greatest alpha diversity between the three sample types for both bacteria and fungi. Dust had the highest total fungal load and the lowest total bacterial load. Sputum had greater bacterial diversity than saliva, but saliva had greater fungal diversity than sputum. The distribution of major bacterial phyla differed between all sample types. However, the distribution of major fungal classes differed only between sputum and saliva. Future research should examine the biological significance of the taxa found in each sample type based on microbial ecology and associations with health effects.

HEPA filtration improves asthma control in children exposed to traffic-related airborne particles.

Traffic-related airborne particles are associated with asthma morbidity. The aim of this study was to assess the impact of a high-efficiency particulate air (HEPA) filtration on the concentrations of traffic particles and the resultant effect on children with asthma. Forty-three children with asthma were enrolled in this double-blind, placebo-controlled crossover design. A HEPA air cleaner or a placebo "dummy" was placed in participants' homes for four weeks, interrupted by a one-month washout period, before crossing over to the other treatment arm for four weeks. Air sampling and health outcomes, including asthma control (ACQ) and quality of life (AQLQ) measures, were completed prior to and at the end of each treatment arm. Indoor concentrations of traffic particles were significantly reduced with the HEPA treatment but not with the "dummy" treatment. In participants with poorly controlled asthma and lower quality of life at baseline, ACQ and AQLQ scores were significantly improved (1.3 to 0.9, P = .003 and 4.9 to 5.5, P = .02, respectively) following the HEPA treatment. In this study, HEPA filtration is associated with improved clinical outcomes and quality of life measures in children with uncontrolled asthma.

Evaluation of personal inhalable aerosol samplers with different filters for use during anthrax responses.

Risk of inhalation exposure to viable Bacillus anthracis (B. anthracis) spores has primarily been assessed using short-term, stationary sampling methods which may not accurately characterize the concentration of inhalable-sized spores reaching a person's breathing zone. While a variety of aerosol sampling methods have been utilized during previous anthrax responses, no consensus has yet been established for personal air sampling. The goal of this study was to determine the best sampler-filter combination(s) for the collection and extraction of B. anthracis spores. The study was designed to (1) evaluate the performance of four filter types (one mixed cellulose ester, MCE (pore size = 3 µm), two polytetrafluoroethylene, PTFE (1 and 3 µm), and one polycarbonate, PC (3 µm)); and (2) evaluate the best performing filters in two commercially available inhalable aerosol samplers (IOM and Button). Bacillus thuringiensis kurstaki [Bt(k)], a simulant for B. anthracis, served as the aerosol challenge. The filters were assessed based on criteria such as ability to maintain low pressure drop over an extended sampling period, filter integrity under various environmental conditions, spore collection and extraction efficiencies, ease of loading and unloading the filters into the samplers, cost, and availability. Three of the four tested collection filters-except MCE-were found suitable for efficient collection and recovery of Bt(k) spores sampled from dry and humid as well as dusty and clean air environments for up to 8 hr. The PC (3 µm) filter was identified as the best performing filter in this study. The PTFE (3 µm) demonstrated a comparable performance, but it is more expensive. Slightly higher concentrations were measured with the IOM inhalable sampler which is the preferred sampler's performance criterion when detecting a highly pathogenic agent with no established "safe" inhalation exposure level. Additional studies are needed to address the effects of environmental conditions and spore concentration. The data obtained in this investigation are crucial for future efforts on the development and optimization of a method for assessing inhalation exposure to B. anthracis.

Culturability of Bacillus spores on aerosol collection filters exposed to airborne combustion products of Al, Mg, and B·Ti.

Destruction of bioweapon facilities due to explosion or fire could aerosolize highly pathogenic microorganisms. The post-event air quality assessment is conducted through air sampling. A bioaerosol sample (often collected on a filter for further culture-based analysis) also contains combustion products, which may influence the microbial culturability and, thus, impact the outcome. We have examined the interaction between spores deposited on collection filters using two simulants of Bacillus anthracis [B. thuringiensis (Bt) and B. atrophaeus (referred to as BG)] and incoming combustion products of Al as well as Mg and B·Ti (common ingredient of metalized explosives). Spores extracted from Teflon, polycarbonate, mixed cellulose ester (MCE), and gelatin filters (most common filter media for bioaerosol sampling), which were exposed to combustion products during a short-term sampling, were analyzed by cultivation. Surprisingly, we observed that aluminum combustion products enhanced the culturability of Bt (but not BG) spores on Teflon filters increasing the culturable count by more than an order of magnitude. Testing polycarbonate and MCE filter materials also revealed a moderate increase of culturability although gelatin did not. No effect was observed with either of the two species interacting on either filter media with products originated by combustion of Mg and B·Ti. Sample contamination, spore agglomeration, effect of a filter material on the spore survival, changes in the spore wall ultrastructure and germination, as well as other factors were explored to interpret the findings. The study raises a question about the reliability of certain filter materials for collecting airborne bio-threat agents in combustion environments.

Key determinants of the fungal and bacterial microbiomes in homes.

BACKGROUND: The microbiome of the home is of great interest because of its possible impact on health. Our goal was to identify some of the factors that determine the richness, evenness and diversity of the home's fungal and bacterial microbiomes. METHOD: Vacuumed settled dust from homes (n=35) in Cincinnati, OH, were analyzed by pyrosequencing to determine the fungal and bacterial relative sequence occurrence. The correlation coefficients between home environmental characteristics, including age of home, Environmental Relative Moldiness Index (ERMI) values, occupant number, relative humidity and temperature, as well as pets (dog and cat) were evaluated for their influence on fungal and bacterial communities. In addition, linear discriminant analysis (LDA) was used for identifying fungal and bacterial genera and species associated with those housing determinants found to be significant. RESULTS: The fungal richness was found to be positively correlated with age of home (p=0.002), ERMI value (p=0.003), and relative humidity (p=0.015) in the home. However, fungal evenness and diversity were only correlated with the age of home (p=0.001). Diversity and evenness (not richness) of the bacterial microbiome in the homes were associated with dog ownership. Linear discriminant analysis showed total of 39 putative fungal genera/species with significantly higher LDA scores in high ERMI homes and 47 genera/species with significantly higher LDA scores in homes with high relative humidity. When categorized according to the age of the home, a total of 67 fungal genera/species had LDA scores above the significance threshold. Dog ownership appeared to have the most influence on the bacterial microbiome, since a total of 130 bacterial genera/species had significantly higher LDA scores in homes with dogs. CONCLUSIONS: Some key determinants of the fungal and bacterial microbiome appear to be excess moisture, age of the home and dog ownership.

Airborne microorganisms, endotoxin, and (1→3)-ß-D-glucan exposure in greenhouses and assessment of respiratory symptoms among workers.

OBJECTIVES: Greenhouse operations are an important sector of the horticulture industry, also known as the Green Industry. The objectives of this study were (i) to investigate exposure levels to airborne culturable fungi, bacteria (total culturable bacteria and actinomycetes), endotoxin, and (1→3)-ß-D-glucan in three Midwest greenhouses during summer and winter using multiple exposure assessment methods; (ii) characterize the load of microorganisms on greenhouse floors and determine potential microbial source strengths of the floors for aerosolizing microbial biocontaminants, and (iii) to estimate the prevalence of rhinitis, wheezing, asthma, and other respiratory symptoms/conditions among greenhouse workers. METHODS: Stationary inhalable aerosol samples were collected from each greenhouse using Button Inhalable Aerosol Samplers. Control samples were collected from offices and nearby outdoor locations. A microbial source strength tester was used to examine the aerosolization potential of microbial contaminants from greenhouse floors. Additionally, surface samples were collected by sterile cotton swabs. Temperature, relative humidity, and wind velocity were recorded. Airborne culturable fungi, bacteria, and actinomycetes were analyzed in the extracts from field samples by cultivation in nutrient agar media. Endotoxin and (1→3)-ß-D-glucan in the extracts from field samples were analyzed by specific kinetic chromogenic Limulus amebocyte lysate assays. The prevalence of respiratory symptoms among greenhouse workers (n = 35) and control subjects (office workers; n = 14) was estimated with a standardized questionnaire. RESULTS AND CONCLUSIONS: The collected data indicate that workers employed in Midwest greenhouses may be exposed to elevated levels of inhalable culturable microorganisms (fungi and bacteria collectively on the order of 10(2)-10(5) CFU m(-3)), endotoxin (10(1)-10(3) EU m(-3)), and (1→3)-ß-D-glucan (10(1)-10(2) ng m(-3)). Seasonal variations were observed for some bioaerosol components. The prevalence of self-reported respiratory symptoms was generally higher among greenhouse workers compared to controls; however, the differences were not statistically significant, likely due to the relatively low statistical power of the study.

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

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

Indoor particulate matter and lung function in children.

People generally spend more time indoors than outdoors resulting in a higher proportion of exposure to particulate matter (PM) occurring indoors. Consequently, indoor PM levels, in contrast to outdoor PM levels, may have a stronger relationship with lung function. To test this hypothesis, indoor and outdoor PM2.5 and fungal spore data were simultaneously collected from the homes of forty-four asthmatic children aged 10-16 years. An optical absorption technique was utilized on the collected PM2.5 mass to obtain concentrations of black carbon (BC) and ultraviolet light absorbing particulate matter, (UVPM; a marker of light absorbing PM2.5 emitted from smoldering organics). Enrolled children completed spirometry after environmental measurements were made. Given the high correlation between PM2.5, BC, and UVPM, principal component analysis was used to obtain uncorrelated summaries of the measured PM. Separate linear mixed-effect models were developed to estimate the association between principal components of the PM variables and spirometry values, as well as the uncorrelated original PM variables and spirometry values. A one-unit increase in the first principal component variable representing indoor PM (predominantly composed of UVPM and PM2.5) was associated with 4.1% decrease (99% CI = -6.9, -1.4) in FEV1/FVC ratio. 11.3 µg/m3 increase in indoor UVPM was associated with 6.4% and 14.7% decrease (99% CI = -10.4, -2.4 and 99% CI = -26.3, -2.9, respectively) in percent predicted FEV1/FVC ratio and FEF25-75 respectively. Additionally, 17.7 µg/m3 increase in indoor PM2.5 was associated with 6.1% and 12.9% decrease (99% CI = -10.2, -1.9 and 99% CI = -24.9, -1.0, respectively) in percent predicted FEV1/FVC ratio and FEF25-75, respectively. Outdoor PM, indoor BC, and indoor fungal spores were not significantly associated with lung function. The results indicate that indoor PM is more strongly associated with lung function in children with asthma as compared with outdoor PM.

Variability of indoor fungal microbiome of green and non-green low-income homes in Cincinnati, Ohio.

"Green" housing is designed to use low-impact materials, increase energy efficiency and improve occupant health. However, little is known about the indoor mycobiome of green homes. The current study is a subset of a multicenter study that aims to investigate the indoor environment of green homes and the respiratory health of asthmatic children. In the current study, the mycobiome in air, bed dust and floor dust was compared between green (study site) and non-green (control site), low-income homes in Cincinnati, Ohio. The samples were collected at baseline (within four months following renovation), and 12months after the baseline at the study site. Parallel sample collection was conducted in non-green control homes. Air samples were collected by PM2.5 samplers over 5-days. Bed and floor dust samples were vacuumed after the air sampling was completed. The DNA sample extracts were analyzed using ITS amplicon sequencing. Analysis indicated that there was no clear trend in the fungal communities between green and non-green homes. Instead, fungal community differences were greatest between sample types - air, bed, and floor. Microbial communities also changed substantially between sampling intervals in both green and non-green homes for all sample types, potentially indicating that there was very little stability in the mycobiomes. Research gaps remain regarding how indoor mycobiome fluctuates over time. Longer follow-up periods might elucidate the effect of green renovation on microbial load in buildings.

Comparison of indoor air sampling and dust collection methods for fungal exposure assessment using quantitative PCR.

Evaluating fungal contamination indoors is complicated because of the many different sampling methods utilized. In this study, fungal contamination was evaluated using five sampling methods and four matrices for results. The five sampling methods were a 48 hour indoor air sample collected with a Button™ inhalable aerosol sampler and four types of dust samples: a vacuumed floor dust sample, newly settled dust collected for four weeks onto two types of electrostatic dust cloths (EDCs) in trays, and a wipe sample of dust from above floor surfaces. The samples were obtained in the bedrooms of asthmatic children (n = 14). Quantitative polymerase chain reaction (qPCR) was used to analyze the dust and air samples for the 36 fungal species that make up the Environmental Relative Moldiness Index (ERMI). The results from the samples were compared by four matrices: total concentration of fungal cells, concentration of fungal species associated with indoor environments, concentration of fungal species associated with outdoor environments, and ERMI values (or ERMI-like values for air samples). The ERMI values for the dust samples and the ERMI-like values for the 48 hour air samples were not significantly different. The total cell concentrations of the 36 species obtained with the four dust collection methods correlated significantly (r = 0.64-0.79, p < 0.05), with the exception of the vacuumed floor dust and newly settled dust. In addition, fungal cell concentrations of indoor associated species correlated well between all four dust sampling methods (r = 0.68-0.86, p < 0.01). No correlation was found between the fungal concentrations in the air and dust samples primarily because of differences in concentrations of Cladosporium cladosporioides Type 1 and Epicoccum nigrum. A representative type of dust sample and a 48 hour air sample might both provide useful information about fungal exposures.

Dustborne and airborne Gram-positive and Gram-negative bacteria in high versus low ERMI homes.

The study aimed at investigating Gram-positive and Gram-negative bacteria in moldy and non-moldy homes, as defined by the home's Environmental Relative Moldiness Index (ERMI) value. The ERMI values were determined from floor dust samples in 2010 and 2011 and homes were classified into low (<5) and high (>5) ERMI groups based on the average ERMI values as well as 2011 ERMI values. Dust and air samples were collected from the homes in 2011 and all samples were analyzed for Gram-positive and Gram-negative bacteria using QPCR assays, endotoxin by the LAL assay, and N-acetyl-muramic acid using HPLC. In addition, air samples were analyzed for culturable bacteria. When average ERMI values were considered, the concentration and load of Gram-positive bacteria determined with QPCR in house dust, but not air, were significantly greater in high ERMI homes than in low ERMI homes. Furthermore, the concentration of endotoxin, but not muramic acid, in the dust was significantly greater in high ERMI than in low ERMI homes. In contrast, when ERMI values of 2011 were considered, Gram-negative bacteria determined with QPCR in air, endotoxin in air, and muramic acid in dust were significantly greater in high ERMI homes. The results suggest that both short-term and long-term mold contamination in homes could be linked with the bacterial concentrations in house dust, however, only the current mold status was associated with bacterial concentrations in air. Although correlations were found between endotoxin and Gram-negative bacteria as well as between muramic acid and Gram-positive bacteria in the entire data set, diverging associations were observed between the different measures of bacteria and the home moldiness. It is likely that concentrations of cells obtained by QPCR and concentrations of cell wall components are not equivalent and represent too broad categories to understand the bacterial composition and sources of the home microbiota.

Visually observed mold and moldy odor versus quantitatively measured microbial exposure in homes.

The main study objective was to compare different methods for assessing mold exposure in conjunction with an epidemiologic study on the development of children's asthma. Homes of 184 children were assessed for mold by visual observations and dust sampling at child's age 1 (Year 1). Similar assessment supplemented with air sampling was conducted in Year 7. Samples were analyzed for endotoxin, (1-3)-ß-D-glucan, and fungal spores. The Mold Specific Quantitative Polymerase Chain Reaction assay was used to analyze 36 mold species in dust samples, and the Environmental Relative Moldiness Index (ERMI) was calculated. Homes were categorized based on three criteria: 1) visible mold damage, 2) moldy odor, and 3) ERMI. Even for homes where families had not moved, Year 7 endotoxin and (1-3)-ß-d-glucan exposures were significantly higher than those in Year 1 (p<0.001), whereas no difference was seen for ERMI (p=0.78). Microbial concentrations were not consistently associated with visible mold damage categories, but were consistently higher in homes with moldy odor and in homes that had high ERMI. Low correlations between results in air and dust samples indicate different types or durations of potential microbial exposures from dust vs. air. Future analysis will indicate which, if any, of the assessment methods is associated with the development of asthma.