Autoclave sterilization and ethanol treatment of re-used surgical masks and N95 respirators during COVID-19: impact on their performance and integrity.

BACKGROUND: An exceptionally high demand for surgical masks and N95 filtering facepiece respirators (FFRs) during the COVID-19 pandemic has considerably exceeded their supply. These disposable devices are generally not approved for routine decontamination and re-use as a standard of care, while this practice has widely occurred in hospitals. The US Centers for Disease Control and Prevention allowed it "as a crisis capacity strategy". However, limited testing was conducted on the impact of specific decontamination methods on the performance of N95 FFRs and no data was presented for surgical masks. AIM: We evaluated common surgical masks and N95 respirators with respect to the changes in their performance and integrity resulting from autoclave sterilization and a 70% ethanol treatment; these methods are frequently utilized for re-used filtering facepieces in hospitals. METHODS: The filter collection efficiency and pressure drop were determined for unused masks and N95 FFRs, and for those subjected to the treatments in a variety of ways. The collection efficiency was measured for particles of approximately 0.037-3.2 µm to represent aerosolized single viruses, their agglomerates, bacteria and larger particle carriers. FINDINGS: The initial collection efficiency and the filter breathability may be compromised by sterilization in an autoclave and ethanol treatment. The effect depends on a protective device, particle size, breathing flow rate, type of treatment and other factors. Additionally, physical damages were observed in N95 respirators after autoclaving. CONCLUSION: Strategies advocating decontamination and re-use of filtering facepieces in hospitals should be re-assessed considering the data obtained in this study.

Concentration gradient patterns of traffic and non-traffic-generated fine and coarse aerosol particles.

The research project described in this article was undertaken to establish baseline information for a Health Impact Assessment (HIA) project of Interstate 75 road construction in Cincinnati, Ohio. The objective of the authors' study was to evaluate the concentrations of elemental and organic carbon (EC and OC), as well as characterize particle number concentrations using devices that measure the fine fraction in the range of 0.02-1 microm and the coarse fraction up to 20 pm. The measurements were conducted at two sites located in the proximity of an interstate highway (at 124 and 277 m) as well as at a remote control site (at >2000 m from any interstate highway). Samples were collected for 24 hours over 12 days in each season (i.e., summer, fall, and winter). Wind data were obtained from the area weather station. Data were analyzed using mixed linear models. Significant increases in concentrations of EC, OC, and fine particles as well as in EC/OC ratios were observed with decreased distance to the highway; this difference was more pronounced in the fall. These results suggest that residents and workers in areas near high-traffic highways may be exposed to elevated levels of airborne fine particles. The results can be used as a baseline for future HIAs of road construction in the area.

Stenotrophomonas, Mycobacterium, and Streptomyces in home dust and air: associations with moldiness and other home/family characteristics.

Respiratory illnesses have been linked to children's exposures to water-damaged homes. Therefore, understanding the microbiome in water-damaged homes is critical to preventing these illnesses. Few studies have quantified bacterial contamination, especially specific species, in water-damaged homes. We collected air and dust samples in twenty-one low-mold homes and twenty-one high-mold homes. The concentrations of three bacteria/genera, Stenotrophomonas maltophilia, Streptomyces sp., and Mycobacterium sp., were measured in air and dust samples using quantitative PCR (QPCR). The concentrations of the bacteria measured in the air samples were not associated with any specific home characteristic based on multiple regression models. However, higher concentrations of S. maltophilia in the dust samples were associated with water damage, that is, with higher floor surface moisture and higher concentrations of moisture-related mold species. The concentrations of Streptomyces and Mycobacterium sp. had similar patterns and may be partially determined by human and animal occupants and outdoor sources of these bacteria.

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.

Evaluation of ionic air purifiers for reducing aerosol exposure in confined indoor spaces.

UNLABELLED: Numerous techniques have been developed over the years for reducing aerosol exposure in indoor air environments. Among indoor air purifiers of different types, ionic emitters have gained increasing attention and are presently used for removing dust particles, aeroallergens and airborne microorganisms from indoor air. In this study, five ionic air purifiers (two wearable and three stationary) that produce unipolar air ions were evaluated with respect to their ability to reduce aerosol exposure in confined indoor spaces. The concentration decay of respirable particles of different properties was monitored in real time inside the breathing zone of a human manikin, which was placed in a relatively small (2.6 m3) walk-in chamber during the operation of an ionic air purifier in calm air and under mixing air condition. The particle removal efficiency as a function of particle size was determined using the data collected with a size-selective optical particle counter. The removal efficiency of the more powerful of the two wearable ionic purifiers reached about 50% after 15 min and almost 100% after 1.5 h of continuous operation in the chamber under calm air conditions. In the absence of external ventilation, air mixing, especially vigorous one (900 CFM), enhanced the air cleaning effect. Similar results were obtained when the manikin was placed inside a partial enclosure that simulated an aircraft seating configuration. All three stationary ionic air purifiers tested in this study were found capable of reducing the aerosol concentration in a confined indoor space. The most powerful stationary unit demonstrated an extremely high particle removal efficiency that increased sharply to almost 90% within 5-6 min, reaching about 100% within 10-12 min for all particle sizes (0.3-3 microm) tested in the chamber. For the units of the same emission rate, the data suggest that the ion polarity per se (negative vs. positive) does not affect the performance but the ion emission rate does. The effects of particle size (within the tested range) and properties (NaCl, PSL, Pseudomonas fluorescens bacteria) as well as the effects of the manikin's body temperature and its breathing on the ionic purifier performance were either small or insignificant. The data suggest that the unipolar ionic air purifiers are particularly efficient in reducing aerosol exposure in the breathing zone when used inside confined spaces with a relatively high surface-to-volume ratio. PRACTICAL IMPLICATIONS: Ionic air purifiers have become increasingly popular for removing dust particles, aeroallergens and airborne microorganisms from indoor air in various settings. While the indoor air cleaning effect, resulting from unipolar and bipolar ion emission, has been tested by several investigators, there are still controversial claims (favorable and unfavorable) about the performance of commercially available ionic air purifiers. Among the five tested ionic air purifiers (two wearable and three stationary) producing unipolar air ions, the units with a higher ion emission rate provided higher particle removal efficiency. The ion polarity (negative vs. positive), the particle size (0.3-3 microm) and properties (NaCl, PSL, Pseudomonas fluorescens bacteria), as well as the body temperature and breathing did not considerable affected the ionization-driven particle removal. The data suggest that the unipolar ionic air purifiers are particularly efficient in reducing aerosol exposure in the breathing zone when they are used inside confined spaces with a relatively high surface-to-volume ratio (such as automobile cabins, aircraft seating areas, bathrooms, cellular offices, small residential rooms, and animal confinements). Based on our experiments, we proposed that purifiers with a very high ion emission rate be operated in an intermittent mode if used indoors for extended time periods. As the particles migrate to and deposit on indoor surfaces during the operation of ionic air purifiers, some excessive surface contamination may occur, which introduces the need of periodic cleaning these surfaces.

Assessment of the aerosolization potential for fungal spores in moldy homes.

UNLABELLED: The airborne fungal concentration measured with air samplers during specific time intervals may not adequately represent the indoor air quality because of the sporadic nature of spore release from sources. The conventional source evaluation (e.g. swab and tape sampling) characterizes the mold source but does not relate to the fraction of spores that can be aerosolized from a contaminated material. As an alternative to these methods, we have recently developed and laboratory-tested a novel Fungal Spore Source Strength Tester (FSSST). It allows assessing the potential of aerosolization of fungal spores from contaminated surfaces under the most favorable release conditions. In this study, the FSSST was used to characterize the release of spores from four building materials in mold-problem homes. The spores of different species were efficiently aerosolized by the FSSST, exhibiting a total spore release rate ranging approximately from 10(2) to 10(3) cm2/min. For all tested materials, <2% of the spores on the contaminated surface were released during the tests. The airborne spore concentration estimated from the release rate data was found in most cases to be significantly greater than the concentration actually measured in these environments with simultaneous air sampling. The results suggest that the FSSST can be used for the assessment of maximum potential exposure to airborne spores released from identified sources in homes. PRACTICAL IMPLICATIONS: A recently developed FSSST was found to be suitable to measure the aerosolization potential of indoor fungal sources at the most favorable release conditions. The FSSST generates the data that allows assessing the strength of mold sources in homes with respect to their maximum ability to contaminate indoor air with fungi. The novel approach bridges two conventional methods, the air sampling and the direct source evaluation (e.g. swab sampling), thus providing a better representation of the airborne fungal exposure than these methods individually. The device prototype can be used for evaluating the effectiveness of environmental interventions by taking samples before and after the intervention. As a broader application, the FSSST can be utilized for assessing the release of various hazardous biological and non-biological particles from contaminated surfaces.

Performance of personal inhalable aerosol samplers in very slowly moving air when facing the aerosol source.

While personal aerosol samplers have been characterized primarily based on wind tunnel tests conducted at relatively high wind speeds, modern indoor occupational environments are usually represented by very slow moving air. Recent surveys suggest that elevated levels of occupational exposure to inhalable airborne particles are typically observed when the worker, operating in the vicinity of the dust source, faces the source. Thus, the first objective of this study was to design and test a new, low cost experimental protocol for measuring the sampling efficiency of personal inhalable aerosol samplers in the vicinity of the aerosol source when the samplers operate in very slowly moving air. In this system, an aerosol generator, which is located in the centre of a room-sized non-ventilated chamber, continuously rotates and omnidirectionally disperses test particles of a specific size. The test and reference samplers are equally distributed around the source at the same distance from the centre and operate in parallel (in most of our experiments, the total number of simultaneously operating samplers was 15). Radial aerosol transport is driven by turbulent diffusion and some natural convection. For each specific particle size and the sampler, the aerosol mass concentration is measured by weighing the collection filter. The second objective was to utilize the new protocol to evaluate three widely used aerosol samplers: the IOM Personal Inhalable Sampler, the Button Personal Inhalable Aerosol Sampler and the 25 mm Millipore filter holder (closed-face C25 cassette). The sampling efficiencies of each instrument were measured with six particle fractions, ranging from 6.9 to 76.9 micro m in their mass median aerodynamic diameter. The Button Sampler efficiency data demonstrated a good agreement with the standard inhalable convention and especially with the low air movement inhalabilty curve. The 25 mm filter holder was found to considerably under-sample the particles larger than 10 micro m; its efficiency did not exceed 7% for particles of 40-100 micro m. The IOM Sampler facing the source was found to over-sample compared with the data obtained previously with a slowly rotating, freely suspended sampler in a low air movement environment. It was also found that the particle wall deposition in the IOM metallic cartridge was rather significant and particle size-dependent. For each sampler (IOM, Button and C25) the precision was characterized through the relative standard deviation (RSD) of the aerosol concentration obtained with identical samplers in a specific experiment. The average RSD was 14% for the IOM Sampler, 11% for the Button Sampler and 35% for the 25 mm filter cassette. A separate set of experiments, performed with the Simplified Torso showed that in very slowly moving air a personal sampler can be adequately evaluated even when it is not attached to a body but freely suspended (confirming the data reported previously).

Limitations of monoclonal antibodies for monitoring of fungal aerosols using Penicillium brevicompactum as a model fungus.

Molds are ubiquitous in every environment and many species have been recently associated with an increase in opportunistic infections in immunocompromised patients or the exacerbation of asthmatic episodes in allergic patients. The degree of environmental contamination with fungi thus needs to be monitored and in this study we report the development of a monoclonal antibody (mAb)-mediated enzyme-linked immunosorbent assay (ELISA) for the detection of spores of Penicillium brevicompactum in experimental model aerosols. In addition, we have investigated the influence of different parameters of air sampling and sample recovery on ELISA performance. MAbs were produced with standard hybridoma techniques and cross-reactivities were determined against spores of 53 fungal species by indirect ELISA. Standardized experimental fungal aerosols were collected with the Button Personal Inhalable Aerosol Sampler onto polycarbonate or polytetrafluoroethylene filters (PTFE) and the effects of different extraction buffers and filter agitation methods during sample processing on spore recovery and ELISA detection were investigated. Five mAbs were produced and all of them cross-reacted with several of 31 related Aspergillus, Penicillium and Eurotium species. However, cross-reactivities with 21 non-related fungi were rare. Spores were recovered in much higher numbers from polycarbonate filters (PFs) than from polytetrafluoroethylene filters. Optical densities (ODs) in ELISA were higher for spores collected into carbonate coating buffer (CCB) than phosphate-buffered saline (PBS). Filter bath sonication following filter vortexing had no positive effects on ELISA sensitivity. The cross-reactivity patterns of mAbs suggest that Aspergillus and Penicillium species share multiple antigens. Quantitative ELISA results for fungal aerosols were found to be influenced by differential sample processing and thus method standardization will be essential to maintain the comparability of immunometric monitoring results.

Comparison of filter bag, cyclonic, and wet dust collection methods in vacuum cleaners.

In this study, methods were developed for comparative evaluation of three primary dust collection methods employed in vacuum cleaners: filter bag, cyclonic, and wet primary dust collection. The dry collectors were evaluated with KCl test aerosols that are commonly used in filter testing. However, these aerosols cannot be used for evaluating wet collectors due to their hygroscopicity. Therefore, the wet collectors were evaluated with nonhygroscopic test particles. Both types of test aerosol indicated similar collection efficiencies in tests with dry collectors. The data show that high initial collection efficiency can be achieved by any one of the three dust collection methods: up to 50% for 0.35 microm particles, and close to 100% for 1.0 microm and larger particles. The degree of dependence of the initial collection efficiency on airflow rate was strongly related to the type and manufacturing of the primary dust collector. Collection efficiency decreased most with decreasing flow rate for the tested wet collectors. The tested cyclonic and wet collectors showed high reentrainment of already collected dust particles. After the filter bag collectors had been loaded with test dust, they also reemitted particles. The degree of reentrainment from filter bags depends on the particulate load and the type of filter material used. Thus, the overall particle emissions performance of a vacuum cleaner depends not only on the dust collection efficiency of the primary collector and other filtration elements employed, but also on the degree of reentrainment of already collected particles.

Particle emission characteristics of filter-equipped vacuum cleaners.

Industrial vacuum cleaners with final high-efficiency particulate air (HEPA) filters traditionally have been used for cleanup operations in which all of the nozzle-entrained dust must be collected with high efficiency, for example, after lead-based paint abatement in homes. In this study household vacuum cleaners ranging from $70 to $650 and an industrial vacuum cleaner costing more than $1400 were evaluated relative to their collection efficiency immediately after installing new primary dust collectors in them. Using newly developed testing technology, some of the low-cost household vacuum cleaners equipped with a final HEPA filter were found to have initial overall filtration efficiencies comparable to those of industrial vacuum cleaners equipped with a final HEPA filter. The household vacuum cleaners equipped with a final HEPA filter efficiently collect about 100% of the dry dust entrained by the nozzle. For extensive cleaning efforts and for vacuum cleaning of wet surfaces, however, industrial vacuum cleaners may have an advantage, including ruggedness and greater loading capacity. The methods and findings of this study are applicable to field evaluations of vacuum cleaners.

Test methods for evaluating the filtration and particulate emission characteristics of vacuum cleaners.

The overall filtration efficiency of a vacuum cleaner traditionally has been tested by placing the vacuum cleaner in a test chamber and measuring aerosol concentrations at the chamber inlet and outlet. The chamber test method was refined and validated in this study. However, this chamber test method shows an overall filtration efficiency of close to 100% for most of the industrial vacuum cleaners and for most of the newly developed household vacuum cleaners of midprice range or higher because all these vacuum cleaners have a high-efficiency particulate air (HEPA) or other highly efficient filter installed at the exhaust. A new test method was therefore developed through which the vacuum cleaner was probed in various internal locations so that the collection efficiency of the individual components could be determined. For example, the aerosol concentration upstream of the final HEPA filter can thus be measured, which permits one to estimate the life expectancy of this expensive component. The probed testing method is particularly suitable for field evaluations of vacuum cleaners because it uses compact, battery-operated optical particle size spectrometers with internal data storage. Both chamber and probed tests gave the same results for the aerosol filtration efficiency. The probed testing method, however, also gives information on the performance of the individual components in a vacuum cleaner. It also can be used to determine the dust pickup efficiency and the degree of reaerosolization of particles collected in the vacuum cleaner.

Performance characteristics of the button personal inhalable aerosol sampler.

The button inhalable aerosol sampler with a curved porous inlet recently was developed and evaluated as a stationary sampler in the laboratory and in the field. The present study focused on investigating its suitability for personal inhalable aerosol sampling. The button sampler was tested at two wind velocities (0.5 and 2.0 m/sec), three particle sizes (7, 29, and 70 microm) and three orientations to the wind (0, 90, and 180 degrees). The performance characteristics of the button sampler were compared with those of three other personal samplers--the IOM (Institute of Occupational Medicine), GSP, and 37-mm closed-face filter cassette. The experiments were conducted in a wind tunnel with the samplers mounted on a full-size manikin. The direction-specific sampling efficiency of the button sampler was found to be essentially independent of the wind direction and dependent on the wind velocity to a much smaller degree than that of the three other samplers. When direction-averaged, the fit of its sampling efficiency curve to the inhalability curve was found to be better than that of the 37-mm closed-face cassette, comparable with that of the GSP sampler, and less than that of the IOM sampler. The precision of the button sampler was found to be generally equal to or better than the precision of the comparison samplers. It was concluded that the button sampler can be successfully used as a personal inhalable aerosol sampler.

Particle settling after lead-based paint abatement work and clearance waiting period.

This study investigated the evolution of airborne particle concentration and size distribution following abatement work in a controlled environment utilizing direct real-time particle monitoring and used it to project potential lead loadings as those particles settle. An 860 ft3 environmental test chamber with sophisticated ventilation and air purifying systems was built. Wooden doors with lead-based paint were dry sanded or scraped to generate the highest feasible airborne lead concentrations. Size-fractional airborne particle concentrations decreased exponentially with time in all tests, even with no air exchange, consistent with the stirred model of constantly mixed air, which predicts longer settling than for tranquil settling. Very low levels of air mixing generated by temperature gradients and initial room air turbulence affected particle settling. About 90% of airborne lead mass settled within 1 hour after active abatement, before final cleaning began. During the second waiting period of 1 hour, which followed cleaning of the floor, additional dust settled so that the additional potential lead loading from remaining airborne lead was less than 20 microg/ft2. For this worst case scenario, the underestimate of the lead loading done by the clearance sampling did not exceed about 30%. For more realistic conditions, the underestimates are projected to be much lower than the new 40 microg/ft2 Housing and Urban Development (HUD) clearance standards for floor dust lead. These results were obtained for the first waiting period (between the end of active abatement and the beginning of cleaning) of 1 hour, as recommended by HUD guidelines. Thus, this study demonstrates no need to increase either the first or second waiting period.

Performance of Air-O-Cell, Burkard, and Button Samplers for total enumeration of airborne spores.

Performance of three devices used for the total enumeration of airborne spores-the Air-O-Cell sampling cassette, the Burkard personal volumetric air sampler, and the Button Aerosol Sampler--was evaluated under controlled laboratory conditions. The first two are glass-slide impactors; the third collects spores on a filter. The samplers were challenged with 0.44-5.10 microm polystyrene latex particles and five microorganisms of 0.84-3.07 microm mean aerodynamic diameter: Streptomyces albus, Bacillus subtilis, Cladosporium cladosporioides, Penicillium brevicompactum, and Penicillium melinii. An optical particle counter measured the particle concentrations upstream and downstream of each sampler, and thus determined the physical collection efficiency of the three samplers. Collection efficiency of the Button Aerosol Sampler was close to 100% for the entire particle size range studied. The cut-off size of each impactor was 2.3-2.4 microm. Acridine orange (with epifluorescent microscopy) and lactophenol cotton blue (with bright light microscopy) staining techniques were used for the microscopic enumeration of spores. No significant difference in microscopic counts was found (at the 95% significance level) when using these two techniques with the Button Aerosol Sampler filters. When the lactophenol cotton blue staining was used to compare total microbial counts yielded by all three samplers, the Button Sampler showed significantly higher counts for the smaller size microorganisms (B. subtilis and C. cladosporioides). For the larger microorganisms (P. brevicompactum and P. melinii) all three samplers yielded similar results. Uniformity of particle deposition on the collection surface was highest for the Button Aerosol Sampler due to the design of its inlet. Thus, the filter collection method used with the Button Aerosol Sampler is suitable and can be advantageous for the enumeration of total airborne spores.

Survival of mycobacteria on N95 personal respirators.

OBJECTIVES: The overall aim of this study was to investigate the survival and possible growth of Mycobacterium tuberculosis simulant bacteria on respirator filters. METHODS: Mycobacterium smegmatis was used as a biochemical simulant for M tuberculosis. Bacterial survival was tested on National Institute for Occupational Safety and Health-certified N95 respirators from three manufacturers. The first experiments simulated one-time respirator use and subsequent storage for 1 to 9 days under ideal conditions for the growth of mycobacteria: 37 degrees C and 85% relative humidity. The bacteria were loaded on the respirator filters under three different nutritional conditions: in the absence of nutrients; in the presence of human saliva (simulating conditions when the respirator is worn); and in the presence of nutrient broth (for ideal growth potential). The subsequent experiments simulated respirator wear for 2 hours under medium work-load conditions at a breathing rate of 56 L/min. RESULTS: It was found that M smegmatis did not grow on the tested respirators, even when the respirators were stored at temperature, humidity, and nutrition conditions most favorable for microbial growth. However, these bacteria could survive on respirators for up to 3 days during storage. The culturability of M smegmatis was not affected by airflow that simulated the breathing rate associated with medium work-load conditions for 2 hours. CONCLUSIONS: This study shows that M tuberculosis surrogate bacteria collected on a respirator are not able to grow and are able to survive only in ideal (ie, not clinically relevant) conditions. Based on these experiments, we conclude that M tuberculosis is unlikely ever to become an infectious problem in the air again, once it is removed by a respirator.

Characteristics of airborne actinomycete spores.

Airborne actinomycete spores, important contaminants in occupational and residential environments, were studied with respect to their (i) release into the air, (ii) aerodynamic and physical size while airborne, and (iii) survival after collection onto agar with an impactor. Three actinomycete species were selected for the tests to exemplify the three main spore types: Streptomyces albus for arthrospores, Micromonospora halophytica for aleuriospores, and Thermoactinomyces vulgaris for endospores. The results show that the incubation conditions (temperature, time, and nutrients) needed for the development of spores for their release into air are different from the conditions that are needed for colony growth only. Additional drying of M. halophytica and T. vulgaris cultures was needed before spores could be released from the culture. The aerodynamic sizes of the spores, measured with an aerodynamic particle sizer, ranged from 0.57 (T. vulgaris) to 1.28 micron (M. halophytica). The physical sizes of the spores, when measured with a microscope and an image analysis system, were found to be smaller than previously reported in the literature. The relative recovery of the spores on agar media ranged from 0.5 (T. vulgaris) to 35% (S. albus). The results indicate that the culturability of the collected airborne actinomycete spores varies widely and is affected by several variables, such as the species and the sampling flow rate. Therefore, alternatives to commonly used cultivation methods need to be developed for the enumeration of actinomycete spores.

Analysis of airborne actinomycete spores with fluorogenic substrates.

The reactions between seven fluorogenic substrates and different groups of enzymes, esterases, lipases, phosphatases, and dehydrogenases, were studied in a search for a new method for the detection of actinomycete spores. Fluorescence measurement was chosen as a fast and sensitive method for microbial analysis. The focus of the research was on the spores of important air contaminants: Streptomyces albus and Thermoactinomyces vulgaris. For the measurement of the enzymatic activity, the chosen fluorogenic substrate was added to a mixture of spores and nutrient media, and the resulting fluorescence was measured with a spectrofluorometer. Fluorogenic substrates were found to show enzymatic activities even for dormant spores. Comparison of the enzymatic activities of dormant spores with those of vegetative cells showed similarity of the enzymatic profiles but higher activity for vegetative cells. The increase of enzymatic activity from dormant spores to vegetative cells was not linear but fluctuating. The largest fluctuations were found after 4 to 5 h of incubation. The enzymatic activities of S. albus were 10 to 50 times lower than those of T. vulgaris, except for the dehydrogenase activity, which was seven times higher. These results indicate that analysis with fluorogenic substrates has the potential for becoming a fast and sensitive method for the enumeration and identification of airborne actinomycete spores.

Performance of N95 respirators: filtration efficiency for airborne microbial and inert particles.

In 1995 the National Institute for Occupational Safety and Health issued new regulations for nonpowered particulate respirators (42 CFR Part 84). A new filter certification system also was created. Among the new particulate respirators that have entered the market, the N95 respirator is the most commonly used in industrial and health care environments. The filtration efficiencies of unloaded N95 particulate respirators have been compared with those of dust/mist (DM) and dust/fume/mist (DFM) respirators certified under the former regulations (30 CFR Part 11). Through laboratory tests with NaCl certification aerosols and measurements with particle-size spectrometers, N95 respirators were found to have higher filtration efficiencies than DM and DFM respirators and noncertified surgical masks. N95 respirators made by different companies were found to have different filtration efficiencies for the most penetrating particle size (0.1 to 0.3 micron), but all were at least 95% efficient at that size for NaCl particles. Above the most penetrating particle size the filtration efficiency increases with size; it reaches approximately 99.5% or higher at about 0.75 micron. Tests with bacteria of size and shape similar to Mycobacterium tuberculosis also showed filtration efficiencies of 99.5% or higher. Experimental data were used to calculate the aerosol mass concentrations inside the respirator when worn in representative work environments. The penetrated mass fractions, in the absence of face leakage, ranged from 0.02% for large particle distributions to 1.8% for submicrometer-size welding fumes. Thus, N95 respirators provide excellent protection against airborne particles when there is a good face seal.

Field testing of new aerosol sampling method with a porous curved surface as inlet.

A new aerosol sampling method, utilizing a porous curved surface as the sampling inlet, has recently been developed. Previous laboratory evaluations of this method have demonstrated its important features, such as low wind sensitivity and good filter collection uniformity. In this study a prototype incorporating the new method was evaluated in the field as a stationary and personal sampling device. The small sampler, utilizing a 25-mm filter is called the button aerosol sampler and was evaluated for collecting total airborne dust and fungal spores. The study was performed in nine poorly maintained inner-city houses during environmental cleanups at different cleanliness levels. The button sampler was used in parallel with the standard 37-mm closed-face filter cassette. Four collocated samplers of each type were tested at all sites as stationary samplers, and three samplers of each type were tested at two sites as personal samplers. Aerosol samples were collected on filters and analyzed using the gravimetric method for total dust and epifluorescence microscopy for fungal spores. The average particle concentration values measured with the button sampler and with the standard sampling cassette were found to correlate well within ranges of 10(1)-10(3) micrograms/m3 for total dust and 10(3)-10(5) spores/m3 for airborne fungi. The measurement results obtained with the new button sampler showed lower intersample variations of the measured concentration levels and higher uniformity of the particle deposits on the filters than those obtained with the standard cassette.

Performance of N95 respirators: reaerosolization of bacteria and solid particles.

If a respirator does not contain an exhalation value, and the respirator wearer sneezes or coughs, one may expect previously collected particles to be reaerosolized. This may be of special concern in environments contaminated with airborne microorganisms. The percentages of reaerosolization were measured in a test setup where the number of reaerosolized particles were registered by dynamic aerosol size spectrometry relative to the number of previously collected particles or bacteria. Experiments at low relative humidity have shown that the reaerosolization of particles below 1 micron, including Mycobacterium tuberculosis surrogate bacteria, does not exceed 0.025%, even if the re-entrainment air velocity is as high as 300 cm/sec (i.e., 37 times the air velocity through the respirator during breathing under heavy workload conditions). The reaerosolization of larger particles into dry air was significant at the highest re-entrainment velocity of 300 cm/sec, which simulates violent sneezing or coughing: 0.1% for 3 microns and about 6% for 5-micron test particles. No reaerosolization was detected at relative humidity levels exceeding 35% at these conditions. Thus, it is concluded that the reaerosolization of particles and bacteria, collected on the fibrous filters of N95 respirators, is insignificant at conditions encountered in respirator wear.