The effect of filter material on bioaerosol collection of Bacillus subtilis spores used as a Bacillus anthracis simulant

The objective of this study was to determine filter materials and extraction methods that are appropriate to use for environmental sampling of B. anthracis. Four types of filters were tested: mixed cellulose ester (MCE) with a pore size of 3 microm, polytetrafluoroethylene (PTFE) with pore sizes of 1 and 3 microm, and gelatin with a pore size of 3 microm. Bacillus subtilis var. niger endospores (also known as Bacillus globigii[BG]) were used as a surrogate for B. anthracis. Endospores were collected into Button Inhalable Aerosol Samplers with sampling times of 15 minutes, 1 hour, and 4 hours. Physical collection efficiency was determined by measuring upstream and downstream B. subtilis concentrations with an optical particle counter. Vortexing with ultrasonic agitation and vortexing with shaker agitation extraction methods were evaluated. The MCE, 1 microm PTFE, and gelatin filters provided physical collection efficiencies of 94% or greater. The 3 microm PTFE filter showed inconsistent physical efficiency characteristics between filters. Epifluorescence microscopic analysis of the gelatin filter extraction fluid revealed the presence of contamination by non-culturable bacteria. Mean differences for microbial culturability were not statistically significant for filter materials and extraction methods. However, the vortexing with shaker agitation extraction method resulted in higher total microbial counts in the extraction fluids for MCE and 1 microm PTFE filters when compared to vortexing with ultrasonic agitation. In summary, the MCE and 1 microm PTFE filters in combination with vortexing and shaker extraction demonstrated the best performance for the filter collection and extraction of BG spores.     

Effect of gaseous chlorine dioxide on indoor microbial contaminants

Traditional and modern techniques for bioaerosol enumeration were used to evaluate the relative efficiency of gaseous chlorine dioxide (ClO2) in reducing the indoor microbial contamination under field and laboratory conditions. The field study was performed in a highly microbially contaminated house, which had had an undetected roof leak for an extended period of time and exhibited large areas of visible microbial growth. Air concentrations of culturable fungi and bacteria, total fungi determined by microscopic count and polymerase chain reaction (PCR) assays, endotoxin, and (1 --> 3)-beta-D-glucan were determined before and after the house was tented and treated with ClO2. The laboratory study was designed to evaluate the efficiency of ClO2 treatment against known concentrations of spores of Aspergillus versicolor and Stachybotrys chartarum on filter paper (surrogate for surface treatment). These species are commonly found in damp indoor environments and were detected in the field study. Upon analysis of the environmental data from the treated house, it was found that the culturable bacteria and fungi as well as total count of fungi (as determined by microscopic count and PCR) were decreased at least 85% after the ClO2 application. However, microscopic analyses of tape samples collected from surfaces after treatment showed that the fungal structures were still present on surfaces. There was no statistically significant change in airborne endotoxin and (1 --> 3)-beta-D-glucan concentration in the field study. The laboratory study supported these results and showed a nonsignificant increase in the concentration of (1 --> 3)-beta-D-glucan after ClO2 treatment.     

Estimating sampling frequency in pollen exposure assessment over time

A time series model was fitted to the pollen concentration data collected in the Greater Cincinnati area for the Cincinnati Childhood Allergy and Air Pollution Study (CCAAPS). A traditional time series analysis and temporal variogram approach were applied to the regularly spaced databases (collected in 2003) and irregularly spaced ones (collected in 2002), respectively. The aim was to evaluate the effect of the sampling frequency on the sampling precision in terms of inverse of standard error of the overall level of mean value across time. The presence of high autocorrelation in the data was confirmed and indicated some degree of temporal redundancy in the pollen concentration data. Therefore, it was suggested that sampling frequency could be reduced from once a day to once every several days without a major loss of sampling precision of the overall mean over time. Considering the trade-offs between sampling frequency and the possibility of sampling bias increasing with larger sampling interval, we recommend that the sampling interval should take values from 3 to 5 days for the pollen monitoring program, if the goal is to track the long-term average.     

Performance of the Button Personal Inhalable Sampler for the measurement of outdoor aeroallergens

No personal aerosol sampler has been evaluated for monitoring aeroallergens in outdoor field conditions and compared to conventional stationary aerobiological samplers. Recently developed Button Personal Inhalable Aerosol Sampler has demonstrated high sampling efficiency for non-biological particles and low sensitivity to the wind direction and velocity. The aim of the present study was to evaluate the Button Sampler for the measurement of outdoor pollen grains and fungal spores side-by-side with the widely used Rotorod Sampler. The sampling was performed for 8 months (spring, summer and fall) at a monitoring station on the roof of a two-storied office building located in the center of the city of Cincinnati. Two identical Button Samplers, one oriented towards the most prevalent wind and the other towards the opposite wind and a Rotorod Sampler were placed side-by-side. The total fungal spore concentration ranged from 129 to 12,980 spores m(-3) (number per cubic meter of air) and the total pollen concentration from 4 to 4536 pollen m(-3). The fungal spore concentrations obtained with the two Button Samplers correlated well (r = 0.95; p<0.0001). The pollen data also showed positive correlation. These findings strongly support the results of earlier studies conducted with non-biological aerosol particles, which demonstrated a low wind dependence of the performance of the Button Sampler compared to other samplers. The Button Sampler's inlet efficiency was found to be more dependent on wind direction when sampling larger sized Pinaceae pollen grains (aerodynamic diameter approximately 65 mum). Compared to Rotorod, both Button Samplers measured significantly higher total fungal spore concentrations. For total pollen count, the Button Sampler facing the prevalent wind showed concentrations levels comparable to that of the Rotorod, but the Button Sampler oriented opposite to the prevalent wind demonstrated lower concentration levels. Overall, it was concluded that the Button Sampler is efficient for the personal sampling of outdoor aeroallergens, and is especially beneficial for aeroallergens of small particle size.     

Culturability and concentration of indoor and outdoor airborne fungi in six single-family homes

In this study, the culturability of indoor and outdoor airborne fungi was determined through long-term sampling (24-h) using a Button Personal Inhalable Aerosol Sampler. The air samples were collected during three seasons in six Cincinnati area homes that were free from moisture damage or visible mold. Cultivation and total microscopic enumeration methods were employed for the sample analysis. The geometric means of indoor and outdoor culturable fungal concentrations were 88 and 102 colony-forming units (CFU) m(-3), respectively, with a geometric mean of the I/O ratio equal to 0.66. Overall, 26 genera of culturable fungi were recovered from the indoor and outdoor samples. For total fungal spores, the indoor and outdoor geometric means were 211 and 605 spores m(-3), respectively, with a geometric mean of I/O ratio equal to 0.32. The identification revealed 37 fungal genera from indoor and outdoor samples based on the total spore analysis. Indoor and outdoor concentrations of culturable and total fungal spores showed significant correlations (r = 0.655, p<0.0001 and r = 0.633, p<0.0001, respectively). The indoor and outdoor median viabilities of fungi were 55% and 25%, respectively, which indicates that indoor environment provides more favorable survival conditions for the aerosolized fungi. Among the seasons, the highest indoor and outdoor culturability of fungi was observed in the fall. Cladosporium had a highest median value of culturability (38% and 33% for indoor and outdoor, respectively) followed by Aspergillus/Penicillium (9% and 2%) among predominant genera of fungi. Increased culturability of fungi inside the homes may have important implications because of the potential increase in the release of allergens from viable spores and pathogenicity of viable fungi on immunocompromised individuals.     

Correlation of ambient inhalable bioaerosols with particulate matter and ozone: a two-year study

In this study, we have examined the relationships between the concentrations of ambient inhalable airborne fungi and pollen with PM10, PM2.5, ozone, organic carbon, selected trace metals (cadmium, copper, lead, and zinc), temperature, and relative humidity. The database was collected in Cincinnati, Ohio, USA, during two consecutive years. Measurements of all environmental variables were performed at the same site continuously 5 days a week except during winter months. The airborne concentrations of biological and non-biological pollutants ranged as follows: total fungi: 184-16 979 spores m(-3); total pollen: 0-6692 pollen m(-3); PM10: 6.70-65.38 microg m(-3); PM2.5: 5.04-45.02 microg m(-3); and ozone: 2.54-64.17 ppb. Higher levels of total inhalable fungi and particulate matter were found during fall and summer months. In contrast, total pollen concentration showed elevated levels in spring. Peak concentrations of ozone were observed during summer and beginning of fall. Our study concluded that several types of inhalable airborne fungi and pollen, particulate matter, and ozone could be positively correlated as a result of the atmospheric temperature influence.     

Airborne rice pollen and pollen allergen in an agricultural field: aerobiological and immunochemical evidence

Grass pollens are well known among the health hazardous bioaerosols causing respiratory allergy. Being an important member of the grass family, the rice plants contribute a huge pollen load in agricultural fields during flowering. This results in a seasonal trigger of hay fever and respiratory allergy in the field workers and people living in the vicinity. Studies on the monitoring of airborne rice pollen and the intensity of the released allergen in agricultural fields are largely lacking. The aims of the present study were: (1) daily and hourly monitoring of airborne rice pollen in an agricultural field during the flowering period of plants in a winter crop season by using the Burkard 7-Day Volumetric Sampler and (2) the measurement of hourly airborne allergen intensity in the field in a peak rice pollen period by the double-antibody and chemiluminescence techniques to find out its relationship with the airborne rice pollen concentration. The monthly average concentration of rice pollen was 95 pollen m(-3) and the range of daily average pollen concentration was 0 to 386 pollen m(-3). A bimodal diurnal periodicity showed that the airborne rice pollen concentrations remained high at two different times of the day: between 08:00 h to 12:00 h and 14:00 h to 16:00 h. Deposition of airborne rice pollen allergen showed darker intensities on the immunostained tapes from the Burkard Sampler at the same two positions corresponding to higher pollen counts. These observations provided direct evidence of the allergenicity of airborne rice pollen in field conditions.     

Volumetric assessment of airborne fungi in two sections of a rural indoor dairy cattle shed

There is increasing concern about the exposure to fungal aerosols in occupational environments and associated respiratory allergic diseases and asthma. A large number of people work in cattle sheds around the world, pulmonary function impairments and higher frequency of respiratory symptoms have been reported in dairy farmers; however, it appears that adequate information on the fungal aerosols from the cattle sheds are largely lacking. Volumetric assessment of airborne culturable and nonculturable fungal spores was performed in two sections of a large rural indoor cattle shed of West Bengal, India for 2 consecutive years. An Andersen Two Stage Viable Sampler was used for sampling culturable fungi and a Burkard Personal Slide Sampler was used to collect the total airborne fungal spores including both the culturable and nonculturable types. A total of 31 spore types and 35 types of viable colony-forming units were recorded. Average concentration range of total fungal spores was 233-2985/m(3) and concentration of viable colony-forming units ranged between 165 and 2225 CFU/m(3). Burkard Sampler showed higher frequencies of Aspergilli/Penicilli, Cladosporium, Alternaria, and smut spores. Andersen Sampler showed the prevalence of Aspergillus niger, Aspergillus flavus and Cladosporium cladosporioides colonies. Some recorded fungi were earlier reported as allergenic, toxic, and pathogenic for occupational workers as well as cattle population. Higher concentration levels of airborne total and culturable fungal spores were recorded during the months of November through February (winter) and June through September (late summer and rainy season).     

Influence of home characteristics on airborne and dustborne endotoxin and β-D-glucan

The aim of this study was to assess the associations between airborne and dustborne microbial contaminants (endotoxin and β-D-glucan) and estimate the effects of home characteristics on exposure levels of these microbial contaminants. Endotoxin and β-D-glucan concentrations in airborne inhalable particles, airborne PM1 (<1 μm) and vacuumed dust from 184 residential homes were determined using specific Limulus amebocyte assays. Home characteristics were recorded by visual inspection and questionnaires. Linear regression and correlation analyses were performed. Inhalable endotoxin correlated with dust endotoxin (r = 0.34, p < 0.001) and PM1 endotoxin (r = 0.33, p < 0.001). Inhalable β-D-glucan correlated with dust β-D-glucan (r = 0.18, p < 0.01), but not with PM1 β-D-glucan. Significant correlation was also found between PM1 and dust concentrations for endotoxin (r = 0.26, p < 0.001), but not for β-D-glucan. Multivariate regression analyses showed only one significant association between airborne contaminants and environmental characteristics: inhalable β-D-glucan was positively associated with relative humidity with an effect size (change in the dependent variable corresponding to a unit increase in the independent variable) of 2.32 and p < 0.05. In contrast, several associations were found between dust concentrations and environmental characteristics. Dust endotoxin was positively associated with temperature (2.87, p < 0.01) and number of inhabitants (2.76, p < 0.01), whereas dust β-D-glucan was inversely associated with the presence of dogs (-2.24, p < 0.05) and carpet (-3.05, p < 0.01) in the home. In conclusion, dustborne contaminants were more strongly affected by home characteristics than airborne contaminants. Furthermore, even though statistically significant, the correlations between airborne and dustborne contaminants were weak. This indicates that airborne concentrations cannot be reliably predicted based on dustborne concentrations.     

A small change in the design of a slit bioaerosol impactor significantly improves its collection characteristics

While several methods are available for bioaerosol monitoring, impaction remains the most common one, particularly for collecting fungal spores. Earlier studies have shown that the collection efficiency of many conventional single-stage bioaerosol impactors falls below 50% for spores with an aerodynamic diameter between 1.7 and 2.5 microm because their cut-off size is 2.5 microm or greater. The cut-off size reduction is primarily done by substantially increasing the sampling flow rate or decreasing the impaction jet size, W, to a fraction of a millimetre, with both measures often impractical to implement. Some success has recently been reported on the utilization of an ultra-low jet-to-plate distance, S (S/W < 0.1), in circular impactors. This paper describes a laboratory evaluation and some field testing of two single-stage, single-nozzle, slit bioaerosol impactors, Allergenco-D and Air-O-Cell, which feature the same jet dimensions and flow rate but have some design configuration differences that were initially thought to be of low significance. The collection efficiency and the spore deposit characteristics were determined in the laboratory using real-time aerosol spectrometry and different microscopic enumeration methods as the test impactors were challenged with the non-biological polydisperse NaCl aerosol and the aerosolized fungal spores of Cladosporium cladosporioides, Aspergillus versicolor, and Penicillium melinii. The tests showed that a relatively small reduction in the jet-to-plate distance of a single-stage, single-nozzle impactor with a tapered inlet nozzle, combined with adding a straight section of sufficient length, can significantly decrease the cut-off size to the level that is sufficient to efficiently collect spores of all fungal species. Furthermore, it appears that the slit jet design may improve the application of partial spore counting methodologies with respect to those applied to circular deposits. Data from a demonstration field study, conducted with the two samplers in environments containing a variety of fungal species, supported the laboratory findings.