Nancy Clark Burton

Physical Collection Efficiency of Filter Materials for Bacteria and Viruses

Abstract The purpose of this study was to determine the physical collection efficiency of commercially available filters for collecting airborne bacteria, viruses, and other particles in the 10–900 nm (nanometer) size range. Laboratory experiments with various polytetrafluoroethylene (PTFE), polycarbonate (PC) and gelatin filters in conjunction with Button™ Inhalable samplers and three-piece cassettes were undertaken. Both biological and non-biological test aerosols were used: Bacillus atrophaeus, MS2, polystyrene latex (PSL), and sodium chloride (NaCl). The B.atrophaeus endospores had an aerodynamic diameter of 900 nm, whereas MS2 virion particles ranged from 10 to 80 nm. Monodisperse 350 nm PSL particles were used as this size was believed to have the lowest filtration efficiency. NaCl solution (1% weight by volume) was used to create a polydisperse aerosol in the 10–600 nm range. The physical collection efficiency was determined by measuring particle concentrations size-selectively upstream and downstream of the filters. The PTFE and gelatin filters showed excellent collection efficiency (>93%) for all of the test particles. The PC filters showed lower collection efficiency for small particles especially <100 nm. Among the tested filters, the lowest collection efficiencies, 49 and 22%, were observed for 1 and 3-μm pore size PC filters at the particle sizes of 47 and 63 nm, respectively. The results indicate that the effect of filter material is more significant for the size range of single virions than for bacteria. The effect of filter loading was examined by exposing filters to mixtures of PSL particles, which aimed at mimicking typical indoor dust levels and size distributions. A 4-h loading did not cause significant change in the physical collection efficiency of the tested filters.

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.

Exposure assessment and analysis for biological agents

Airborne biological agents have become prominent safety and health issues in agriculture, biotechnology, industrial settings, and the indoor environment. Each of these environments presents unique exposure concerns due to the nature of the encountered biological agent, the microbial concentrations, the modes of exposure, and the susceptibility of the exposed population. Acceptable levels of airborne microorganisms have not been established and the sampling methods and analytical techniques employed to assess airborne biocontaminants are varied and non-standardized. This paper reviews and compares the different air sampling methods for biological agents and classical analytical methods (i.e., culture and microscopy), analysis for specific microorganism constituents (i.e., ergosterol, muramic acid, glucans, allergens, mycotoxins, endotoxins) and molecular methods (i.e., polymerase chain reactions). Each of the described methods has distinct advantages and disadvantages. Selection of sampling and analytical methods depends upon the nature of the information that is sought; there is no one ideal sampling or analytical method. Combinations of sampling and analytical methods can provide a wide range of data that can be effectively tailored to many different environmental settings.

Effect of Gaseous Chlorine Dioxide on Indoor Microbial Contaminants

Abstract 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)-β-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)-β-D-glucan concentration in the field study. The laboratory study supported these results and showed a nonsignificant increase in the concentration of (1→3)-β-D-glucan after ClO2 treatment.

Comparison of work-related symptoms and visual contrast sensitivity between employees at a severely water-damaged school and a school without significant water damage.

BACKGROUND The National Institute for Occupational Safety and Health (NIOSH) conducted a health hazard evaluation (HHE) of a water-damaged school in New Orleans (NO), Louisiana. Our aim in this evaluation was to document employee health effects related to exposure to the water-damaged school, and to determine if VCS testing could serve as a biomarker of effect for occupants who experienced adverse health effects in a water-damaged building. METHODS NIOSH physicians and staff administered a work history and medical questionnaire, conducted visual contrast sensitivity (VCS) testing, and collected sticky-tape, air, and dust samples at the school. Counting, culturing, and/or a DNA-based technology, called mold-specific quantitative PCR (MSQPCR), were also used to quantify the molds. A similar health and environmental evaluation was performed at a comparable school in Cincinnati, Ohio which was not water-damaged. RESULTS Extensive mold contamination was documented in the water-damaged school and employees (n = 95) had higher prevalences of work-related rashes and nasal, lower respiratory, and constitutional symptoms than those at the comparison school (n = 110). VCS values across all spatial frequencies were lower among employees at the water-damaged school. CONCLUSIONS Employees exposed to an extensively water-damaged environment reported adverse health effects, including rashes and nasal, lower respiratory, and constitutional symptoms. VCS values were lower in the employees at the water-damaged school, but we do not recommend using it in evaluation of people exposed to mold. Am. J. Ind. Med. 55:844-854, 2012. This article is a U.S. Government work and is in the public domain in the USA.

Microbial hazards during harvesting and processing at an outdoor United States cannabis farm

ABSTRACT Cannabis cultivation is an emerging industry within the United States. Organic dust derived in part from naturally occurring microorganisms is known to cause byssinosis in the hemp industry. In this pilot study, bacteria and fungi encountered by workers at an outdoor cannabis farm that utilized organic practices were elucidated by 16 S ribosomal RNA (rRNA) and Internal Transcribed Spacer (ITS) region sequencing, respectively. Area (n = 14) and personal air samples (n = 12) were collected during harvesting and processing activities. 16 S rRNA and ITS regions of extracted bacterial and fungal genomic DNA were amplified and sequenced using Sanger sequencing. Bacterial sequencing resolved 1,077 sequences that were clustered into 639 operational taxonomic units (OTUs) and predominantly placed in the phylum, Actinobacteria (46%). Personal air samples revealed higher bacterial and Actinobacteria diversity compared to outdoor area samples collected within the facility (p < 0.05). A high degree of dissimilarity between bacteria was identified within and between samples. Fungal sequences (n = 985) were identified and predominantly clustered in the phylum Ascomycota (53%). Of the 216 fungal OTUs elucidated, the cannabis plant pathogenic species, Botrytis cinerea, was the most prevalent and accounted for 34% of all fungal sequences. The relative abundance of B. cinerea was highest in personal air samples (59%) compared to area samples collected in the drying room (19%), greenhouse (18%), and outdoor environment (6%). There was 49% sample similarity between fungi identified within personal air samples, but higher dissimilarity coefficients were observed within and between greenhouse, drying room, and outdoor area air samples. The results of this pilot study suggest that the cannabis farm workers are potentially exposed to Actinobacteria as well as the cannabis plant pathogen, B. cinerea during harvesting, bud-stripping, and hand-trimming processes.

Investigating a persistent odor at an aircraft seat manufacturer

ABSTRACT An aircraft seat manufacturing company requested a NIOSH health hazard evaluation to help identify a strong odor that had persisted throughout the facility for over a year. Employees reported experiencing health effects thought to be related to the odor. We collected and analyzed area air samples for volatile organic compounds, endotoxin, bacterial and fungal metagenome, and metalworking fluid aerosol. Bulk metalworking fluid samples were analyzed for endotoxin, bacterial and fungal metagenome, and viable bacteria and fungus. We also evaluated the building ventilation systems and water diversion systems. Employees underwent confidential medical interviews about work practices, medical history, and health concerns. Based on our analyses, the odor was likely 2-methoxy-3,5-dimethylpyrazine. This pyrazine was found in air samples across the facility and originated from bacteria in the metalworking fluid. We did not identify bacteria known to produce the compound but bacteria from the same Proteobacteria order were found as well as bacteria from orders known to produce other pyrazines. Chemical and biological contaminants and odors could have contributed to health symptoms reported by employees, but it is likely that the symptoms were caused by several factors. We provided several recommendations to eliminate the odor including washing and disinfecting the metalworking machines and metalworking fluid recycling equipment, discarding all used metalworking fluid, instituting a metalworking fluid maintenance program at the site, and physically isolating the metalworking department from other departments.