Impact of sampling and storage stress on the recovery of airborne SARS-CoV-2 virus surrogate captured by filtration.

Environmental air sampling of the SARS-CoV-2 virus in occupational and community settings is pertinent to reduce and monitor the spread of the COVID pandemic. However, there is a general lack of standardized procedures for airborne virus sampling and limited knowledge of how sampling and storage stress impact the recovery of captured airborne viruses. Since filtration is one of the commonly used methods to capture airborne viruses, this study analyzed the effect of sampling and storage stress on SARS-CoV-2 surrogate virus (human coronavirus OC43, or HCoV-OC43) captured by filters. HCoV-OC43, a simulant of the SARS-CoV-2, was aerosolized and captured by PTFE-laminated filters. The impact of sampling stress was evaluated by comparing the RNA yields recovered when sampled at 3 L/min and 10 L/min and for 10 min and 60 min; in one set of experiments, additional stress was added by passing clean air through filters with the virus for 1, 5, and 15 hr. The impact of storage stress was designed to examine RNA recovery from filters at room temperature (25 °C) and refrigerated conditions (4 °C) for up to 1 week of storage. To our knowledge, this is the first report on using HCoV-OC43 aerosol in air sampling experiments, and the mode diameter of the virus aerosolized from the growth medium was 40-60 nm as determined by SMPS + CPC system (TSI Inc.) and MiniWRAS (Grimm Inc.) measurements. No significant difference was found in virus recovery between the two sampling flow rates and different sampling times (p > 0.05). However, storage at room temperature (25 °C) yielded ∼2x less RNA than immediate processing and storage at refrigerated conditions (4 °C). Therefore, it is recommended to store filter samples with viruses at 4 °C up to 1 week if the immediate analysis is not feasible. Although the laminated PTFE filter used in this work purposefully does not include a non-PTFE backing, the general recommendations for handling and storing filter samples with viral particles are likely to apply to other filter types.

Application of end-of-shift respirable crystalline silica monitoring to construction.

A pilot project was conducted to determine the effect of common construction dusts as interferences in a new portable end-of-shift (EoS), direct-on-filter (DoF) sampling and analysis method for respirable crystalline silica (RCS), in this case, quartz. Construction dusts were prepared from plaster, drywall, cement and brick by grinding, aerosolizing, and collecting respirable dust with high flow rate cyclones. Filters were loaded with different levels of commercial α-quartz powder Min-u-Sil 5, and different levels of interfering dusts, singly and in combination. Samples were analyzed by Fourier Transform Infrared Spectroscopy (FTIR). Good correlations were found between nominal quartz loading (0 µg, 25 µg, 50 µg, and 100 µg) adjusted for quartz in the interfering dust and FTIR absorbance alone and in the presence of all interfering dusts. The slopes of the correlations were similar whether the loading was quartz without interference, or with plaster, drywall, and cement dusts, regardless of quantity. The results show that (a) plaster and drywall dusts do not interfere substantially; (b) cement does not interfere, but a change in the intercept suggests an effect on the background absorbance of the filter; and (c) in addition to having a substantial quartz content, brick dust contains an additional material, probably a silicate mineral, which interferes with the quartz peak. Thus, the presence of cement leads to lower quartz values and brick leads to higher values, but overall, 83% of the quartz contents predicted from the calibration data agreed within 50% of the adjusted nominal loadings within the range 20-110 µg. This result is encouraging given the high levels of interfering dusts. Nine samples loaded with smaller amounts of all four dusts together gave results within 25% of the adjusted nominal loadings. A single mixture addition of the dusts to the filter gave tighter variance in results than sequential additions. Unexpectedly, the two Certified Reference Materials (CRMs) 1878a and 1878b, gave different results when used to calibrate XRD analysis of Min-u-Sil 5.

Comparative cytotoxicity of respirable surface-treated/untreated calcium carbonate rock dust particles in vitro.

Calcium carbonate rock dust (RD) is used in mining to reduce the explosivity of aerosolized coal. During the dusting procedures, potential for human exposure occurs, raising health concerns. To improve RD aerosolization, several types of anti-caking surface treatments exist. The aim of the study was to evaluate cytotoxicity of four respirable RD samples: untreated/treated limestone (UL/TL), untreated/treated marble (UM/TM), and crystalline silica (SiO2) as a positive control in A549 and THP-1 transformed human cell lines. Respirable fractions were generated and collected using FSP10 high flow-rate cyclone samplers. THP-1 cells were differentiated with phorbol-12-myristate-13-acetate (20 ng/ml, 48 h). Cells were exposed to seven different concentrations of RD and SiO2 (0-0.2 mg/ml). RD caused a slight decrease in viability at 24 or 72 h post-exposure and were able to induce inflammatory cytokine production in A549 cells, however, with considerably less potency than SiO2. In THP-1 cells at 24 h, there was significant dose-dependent lactate dehydrogenase, inflammatory cytokine and chemokine release. Caspase-1 activity was increased in SiO2- and, on a lesser scale, in TM- exposed cells. To test if the increased toxicity of TM was uptake-related, THP-1 cells were pretreated with Cytochalasin D (CytD) or Bafilomycin A (BafA), followed by exposure to RD or SiO2 for 6 h. CytD blocked the uptake and significantly decreased cytotoxicity of all particles, while BafA prevented caspase-1 activation but not cytotoxic effects of TM. Only TM was able to induce an inflammatory response in THP-1 cells, however it was much less pronounced compared to silica.

Characterization and assessment of the potential toxicity/pathogenicity of fibrous glaucophane.

In California, the metamorphic blueschist occurrences within the Franciscan Complex are commonly composed of glaucophane, which can be found with a fibrous habit. Fibrous glaucophane's potential toxicity/pathogenicity has never been determined and it has not been considered by the International Agency for Research on Cancer (IARC) as a potential carcinogen to date. Notwithstanding, outcrops hosting fibrous glaucophane are being excavated today in California for building/construction purposes (see for example the Calaveras Dam Replacement Project - CDRP). Dust generated by these excavation activities may expose workforces and the general population to this potential natural hazard. In this work, the potential toxicity/pathogenicity of fibrous glaucophane has been determined using the fibre potential toxicity index (FPTI). This model has been applied to a representative glaucophane-rich sample collected at San Anselmo, Marin County (CA, USA), characterized using a suite of experimental techniques to determine morphometric, crystal-chemical parameters, surface reactivity, biodurability and related parameters. With respect to the asbestos minerals, the FPTI of fibrous glaucophane is remarkably higher than that of chrysotile, and comparable to that of tremolite, thus supporting the application of the precautionary approach when excavating fibrous glaucophane-rich blueschist rocks. Because fibrous glaucophane can be considered a potential health hazard, just like amphibole asbestos, it should be taken into consideration in the standard procedures for the identification and assessment of minerals fibres in soil and air samples.

Characterization of pulmonary responses in mice to asbestos/asbestiform fibers using gene expression profiles.

Humans exposed to asbestos and/or asbestiform fibers are at high risk of developing many lung diseases including asbestosis, lung cancer, and malignant mesothelioma. However, the disease-causing potential and specific metabolic mechanisms and pathways associated with various asbestos/asbestiform fiber exposures triggering different carcinogenic and non-carcinogenic outcomes are still largely unknown. The aim of this this study was to investigate gene expression profiles and inflammatory responses to different asbestos/asbestiform fibers at the acute/sub-acute phase that may be related to delayed pathological outcomes observed at later time points. Mice were exposed to asbestos (crocidolite, tremolite asbestos), asbestiform fibers (erionite), and a low pathogenicity mineral fiber (wollastonite) using oropharyngeal aspiration. Similarities in inflammatory and tissue damage responses, albeit with quantitative differences, were observed at day 1 and 7 post treatment. Exposure to different fibers induced significant changes in regulation and release of a number of inflammatory cytokines/chemokines. Comparative analysis of changes in gene regulation in the lung on day 7 post exposure were interpretable in the context of differential biological responses that were consistent with histopathological findings at days 7 and 56 post treatment. Our results noted differences in the magnitudes of pulmonary responses and gene regulation consistent with pathological alterations induced by exposures to four asbestos/asbestiform fibers examined. Further comparative mechanistic studies linking early responses with the long-term endpoints may be instrumental to understanding triggering mechanisms underlying pulmonary carcinogenesis, that is lung cancer versus mesothelioma.

Interlaboratory comparison for the determination of the soluble fraction of metals in welding fume samples.

There is interest in the bioaccessible metal components of aerosols, but this has been minimally studied because standardized sampling and analytical methods have not yet been developed. An interlaboratory study (ILS) has been carried out to evaluate a method for determining the water-soluble component of realistic welding fume (WF) air samples. Replicate samples were generated in the laboratory and distributed to participating laboratories to be analyzed according to a standardized procedure. Within-laboratory precision of replicate sample analysis (repeatability) was very good. Reproducibility between laboratories was not as good, but within limits of acceptability for the analysis of typical aerosol samples. These results can be used to support the development of a standardized test method.

Assessing occupational erionite and respirable crystalline silica exposure among outdoor workers in Wyoming, South Dakota, and Montana.

Erionite is a naturally occurring fibrous mineral found in many parts of the world, including the western United States. Inhalational exposure to erionite fibers in some localities is associated with health effects similar to those caused by asbestos exposure, including malignant mesothelioma. Therefore, there is concern regarding occupational exposures in the western United States. Currently, there are no standard sampling and analytical methods for airborne erionite fibers, as well as no established occupational exposure limits. Due to the potential adverse health effects, characterizing and minimizing exposures is prudent. Crystalline silica also occurs naturally in areas where erionite is found, principally as the mineral quartz. Work activities involving rocks containing quartz and soils derived from those rocks can lead to exposure to respirable crystalline silica (RCS). The typically dry and dusty environment of the western United States can increase the likelihood of exposures to aerosolized rocks and soils, but inhalation exposure is also possible in more humid conditions. In this case study, we describe several outdoor occupational environments with potential exposures to erionite and RCS. We describe our method for evaluating those exposures and demonstrate: (1) the occurrence of occupational exposures to airborne erionite and RCS, (2) that the chemical make-up of the erionite mineral can be determined, and (3) that effective dust control practices are needed to reduce employee exposures to these minerals.

Surgical smoke control with local exhaust ventilation: Experimental study.

This experimental study aimed to evaluate airborne particulates and volatile organic compounds (VOCs) from surgical smoke when a local exhaust ventilation (LEV) system is in place. Surgical smoke was generated from human tissue in an unoccupied operating room using an electrocautery surgical device for 15 min with 3 different test settings: (1) without LEV control; (2) control with a wall irrigation suction unit with an in-line ultra-low penetration air filter; and (3) control with a smoke evacuation system. Flow rate of LEVs was approximately 35 L/min and suction was maintained within 5 cm of electrocautery interaction site. A total of 6 experiments were conducted. Particle number and mass concentrations were measured using direct reading instruments including a condensation particle counter (CPC), a light-scattering laser photometer (DustTrak DRX), a scanning mobility particle sizer (SMPS), an aerodynamic particle sizer (APS), and a viable particle counter. Selected VOCs were collected using evacuated canisters using grab, personal and area sampling techniques. The largest average particle and VOCs concentrations were found in the absence of LEV control followed by LEV controls. Average ratios of LEV controls to without LEV control ranged 0.24-0.33 (CPC), 0.28-0.39 (SMPS), 0.14-0.31 (DustTrak DRX), and 0.26-0.55 (APS). Ethanol and isopropyl alcohol were dominant in the canister samples. Acetaldehyde, acetone, acetonitrile, benzene, hexane, styrene, and toluene were detected but at lower concentrations (<500 µg/m3) and concentrations of the VOCs were much less than the National Institute for Occupational Safety and Health recommended exposure limit values. Utilization of the LEVs for surgical smoke control can significantly reduce but not completely eliminate airborne particles and VOCs.

Evaluation of a portable gas chromatograph with photoionization detector under variations of VOC concentration, temperature, and relative humidity.

The objective of this present study was to evaluate the performance of a portable gas chromatograph-photoionization detector (GC-PID), under various test conditions to determine if it could be used in occupational settings. A mixture of 7 volatile organic compounds (VOCs)-acetone, ethylbenzene, methyl isobutyl ketone, toluene, m-xylene, p-xylene, and o-xylene-was selected because its components are commonly present in paint manufacturing industries. A full-factorial combination of 4 concentration levels (exposure scenarios) of VOC mixtures, 3 different temperatures (25°C, 30°C, and 35°C), and 3 relative humidities (RHs; 25%, 50%, and 75%) was conducted in a full-size controlled environmental chamber. Three repetitions were conducted for each test condition allowing for estimation of accuracy. Time-weighted average exposure data were collected using solid sorbent tubes (Anasorb 747, SKC Inc.) as the reference sampling medium. Calibration curves of Frog-4000 using the dry gases showed R2 > 0.99 for all analytes except for toluene (R2 = 0.97). Frog-4000 estimates within a test condition showed good consistency for the performance of repeated measurement. However, there was ∼41-64% reduction in the analysis of polar acetone with 75% RH relative to collection at 25% RH. Although Frog-4000 results correlated well with solid sorbent tubes (r = 0.808-0.993, except for toluene) most of the combinations regardless of analyte did not meet the <25% accuracy criterion recommended by NIOSH. The effect of chromatographic co-elution can be seen with m, p-xylene when the results are compared to the sorbent tube sampling technique with GC-flame ionization detector. The results indicated an effect of humidity on the quantification of the polar compounds that might be attributed to the pre-concentrator placed in the selected GC-PID. Further investigation may resolve the humidity effect on sorbent trap with micro GC pre-concentrator when water vapor is present. Although this instrument does not fulfill the accuracy criterion specified in the NIOSH technical report No. 2012-162, it can be used as a screening tool for range finding monitoring with dry gases calibration in the occupational setting rather than compliance monitoring.

Respirable size-selective sampler for end-of-shift quartz measurement: Development and performance.

Aims of this study were to develop a respirable size-selective sampler for direct-on-filter (DoF) quartz measurement at the end-of-shift (EoS) using a portable Fourier transform infrared (FTIR) spectrometer and to determine its size-selective sampling performance. A new miniaturized sampler has been designed to have an effective particle deposition diameter close to the portable FTIR beam diameter (6 mm). The new sampler (named the EoS cyclone) was constructed using a 3D printer. The sampling efficiency of the EoS cyclone was determined using polydisperse glass sphere particles and a time-of-flight direct reading instrument. Respirable dust mass concentration and quartz absorbance levels of samples collected with the EoS cyclone were compared to those collected with the 10-mm nylon cyclone. The EoS cyclone operated at a flow rate of 1.2 l min-1 showed minimum bias compared to the international standard respirable convention. The use of the EoS cyclone induced respirable dust mass concentration results similar but significantly larger (5%) than those obtained from samples collected with 10-mm nylon cyclones. The sensitivity of the DoF-FTIR analysis in estimating quartz was found increased more than 10 times when the samples were collected with the EoS cyclone. The average particle deposition diameter was 8.8 mm in 60 samples. The newly developed user friendly EoS cyclone may provide a better sampling strategy in quartz exposure assessment with faster feedback.

Comparison between active (pumped) and passive (diffusive) sampling methods for formaldehyde in pathology and histology laboratories.

This study was to determine occupational exposures to formaldehyde and to compare concentrations of formaldehyde obtained by active and passive sampling methods. In one pathology and one histology laboratories, exposure measurements were collected with sets of active air samplers (Supelco LpDNPH tubes) and passive badges (ChemDisk Aldehyde Monitor 571). Sixty-six sample pairs (49 personal and 17 area) were collected and analyzed by NIOSH NMAM 2016 for active samples and OSHA Method 1007 (using the manufacturer's updated uptake rate) for passive samples. All active and passive 8-hr time-weighted average (TWA) measurements showed compliance with the OSHA permissible exposure limit (PEL-0.75 ppm) except for one passive measurement, whereas 78% for the active and 88% for the passive samples exceeded the NIOSH recommended exposure limit (REL-0.016 ppm). Overall, 73% of the passive samples showed higher concentrations than the active samples and a statistical test indicated disagreement between two methods for all data and for data without outliers. The OSHA Method cautions that passive samplers should not be used for sampling situations involving formalin solutions because of low concentration estimates in the presence of reaction products of formaldehyde and methanol (a formalin additive). However, this situation was not observed, perhaps because the formalin solutions used in these laboratories included much less methanol (3%) than those tested in the OSHA Method (up to 15%). The passive samplers in general overestimated concentrations compared to the active method, which is prudent for demonstrating compliance with an occupational exposure limit, but occasional large differences may be a result of collecting aerosolized droplets or splashes on the face of the samplers. In the situations examined in this study the passive sampler generally produces higher results than the active sampler so that a body of results from passive samplers demonstrating compliance with the OSHA PEL would be a valid conclusion. However, individual passive samples can show lower results than a paired active sampler so that a single result should be treated with caution.

A New Miniature Respirable Sampler for In-mask Sampling: Part 1-Particle Size Selection Performance.

The Health and Safety Laboratory has developed a miniature respirable sampler to gain a better understanding of the exposure of workers to hazardous substances when they are wearing respiratory protective equipment (RPE) or helmets with visors in the workplace. The study was in two parts and the first part, described herein, was to develop the sampler and test its collection characteristics. Assessment of the impact of the sampler on RPE safety and its comparability with traditional laboratory-based approaches to measure protection factors was discussed in a second article. The miniature sampler (weight-5.4g, length-13mm) was designed to fit into the space available between the nose and chin of an individual inside a filtering facepiece type mask and has a radially omnidirectional inlet with a porous foam particle selector that allows the collection of the respirable fraction on a downstream filter. The sampling efficiency was compared with the respirable convention. A close match with the respirable convention was obtained at a flow rate of 1 l min-1 and the 50% penetration cut off value (d 50) was 4.08 µm. After 3 hours sampling in high humidity (95%), the penetration curve had shifted towards smaller particle sizes (d 50 = 3.81 µm) with 88% of the calculated bias values within 10%. The miniature sampler measured respirable dust and crystalline silica mass concentrations comparable with performance of the Safety In Mines Personal Dust Sampler (SIMPEDS), commonly used in Great Britain, at a flow rate of 0.8 l min-1 The d 50 for the miniature sampler at 0.8 l min-1 (4.4 µm) is within 5% of the d 50 of the SIMPEDS at its prescribed flow rate of 2.2 l min-1 (4.2 µm). These results indicated that the miniature sampler was a good candidate to proceed with tests with RPE described in the second part of this series of two papers.

Silica Measurement with High Flow Rate Respirable Size Selective Samplers: A Field Study.

High and low flow rate respirable size selective samplers including the CIP10-R (10 l min(-1)), FSP10 (11.2 l min(-1)), GK2.69 (4.4 l min(-1)), 10-mm nylon (1.7 l min(-1)), and Higgins-Dewell type (2.2 l min(-1)) were compared via side-by-side sampling in workplaces for respirable crystalline silica measurement. Sampling was conducted at eight different occupational sites in the USA and five different stonemasonry sites in Ireland. A total of 536 (268 pairs) personal samples and 55 area samples were collected. Gravimetric analysis was used to determine respirable dust mass and X-ray diffraction analysis was used to determine quartz mass. Ratios of respirable dust mass concentration, quartz mass concentration, respirable dust mass, and quartz mass from high and low flow rate samplers were compared. In general, samplers did not show significant differences greater than 30% in respirable dust mass concentration and quartz mass concentration when outliers (ratio <0.3 or >3.0) were removed from the analysis. The frequency of samples above the limit of detection and limit of quantification of quartz was significantly higher for the CIP10-R and FSP10 samplers compared to low flow rate samplers, while the GK2.69 cyclone did not show significant difference from low flow rate samplers. High flow rate samplers collected significantly more respirable dust and quartz than low flow rate samplers as expected indicating that utilizing high flow rate samplers might improve precision in quartz measurement. Although the samplers did not show significant differences in respirable dust and quartz concentrations, other practical attributes might make them more or less suitable for personal sampling.

Calibration of high flow rate thoracic-size selective samplers.

High flow rate respirable size selective samplers, GK4.126 and FSP10 cyclones, were calibrated for thoracic-size selective sampling in two different laboratories. The National Institute for Occupational Safety and Health (NIOSH) utilized monodisperse ammonium fluorescein particles and scanning electron microscopy to determine the aerodynamic particle size of the monodisperse aerosol. Fluorescein intensity was measured to determine sampling efficiencies of the cyclones. The Health Safety and Laboratory (HSL) utilized a real time particle sizing instrument (Aerodynamic Particle Sizer) and polydisperse glass sphere particles and particle size distributions between the cyclone and reference sampler were compared. Sampling efficiency of the cyclones were compared to the thoracic convention defined by the American Conference of Governmental Industrial Hygienists (ACGIH)/Comité Européen de Normalisation (CEN)/International Standards Organization (ISO). The GK4.126 cyclone showed minimum bias compared to the thoracic convention at flow rates of 3.5 l min(-1) (NIOSH) and 2.7-3.3 l min(-1) (HSL) and the difference may be from the use of different test systems. In order to collect the most dust and reduce the limit of detection, HSL suggested using the upper end in range (3.3 l min(-1)). A flow rate of 3.4 l min(-1) would be a reasonable compromise, pending confirmation in other laboratories. The FSP10 cyclone showed minimum bias at the flow rate of 4.0 l min(-1) in the NIOSH laboratory test. The high flow rate thoracic-size selective samplers might be used for higher sample mass collection in order to meet analytical limits of quantification.

Replacement of filters for respirable quartz measurement in coal mine dust by infrared spectroscopy.

The objective of this article is to compare and characterize nylon, polypropylene (PP), and polyvinyl chloride (PVC) membrane filters that might be used to replace the vinyl/acrylic co-polymer (DM-450) filter currently used in the Mine Safety and Health Administration (MSHA) P-7 method (Quartz Analytical Method) and the National Institute for Occupational Safety and Health (NIOSH) Manual of Analytical Methods 7603 method (QUARTZ in coal mine dust, by IR re-deposition). This effort is necessary because the DM-450 filters are no longer commercially available. There is an impending shortage of DM-450 filters. For example, the MSHA Pittsburgh laboratory alone analyzes annually approximately 15,000 samples according to the MSHA P-7 method that requires DM-450 filters. Membrane filters suitable for on-filter analysis should have high infrared (IR) transmittance in the spectral region 600-1000 cm(-1). Nylon (47 mm, 0.45 µm pore size), PP (47 mm, 0.45 µm pore size), and PVC (47 mm, 5 µm pore size) filters meet this specification. Limits of detection and limits of quantification were determined from Fourier transform infrared spectroscopy (FTIR) measurements of blank filters. The average measured quartz mass and coefficient of variation were determined from test filters spiked with respirable α-quartz following MSHA P-7 and NIOSH 7603 methods. Quartz was also quantified in samples of respirable coal dust on each test filter type using the MSHA and NIOSH analysis methods. The results indicate that PP and PVC filters may replace the DM-450 filters for quartz measurement in coal dust by FTIR. PVC filters of 5 µm pore size seemed to be suitable replacement although their ability to retain small particulates should be checked by further experiment.

Treated and untreated rock dust: Quartz content and physical characterization.

Rock dusting is used to prevent secondary explosions in coal mines, but inhalation of rock dusts can be hazardous if the crystalline silica (e.g., quartz) content in the respirable fraction is high. The objective of this study is to assess the quartz content and physical characteristics of four selected rock dusts, consisting of limestone or marble in both treated (such as treatment with stearic acid or stearates) and untreated forms. Four selected rock dusts (an untreated and treated limestone and an untreated and treated marble) were aerosolized in an aerosol chamber. Respirable size-selective sampling was conducted along with particle size-segregated sampling using a Micro-Orifice Uniform Deposit Impactor. Fourier Transform Infrared spectroscopy and scanning electron microscopy with energy-dispersive X-ray (SEM-EDX) analyses were used to determine quartz mass and particle morphology, respectively. Quartz percentage in the respirable dust fraction of untreated and treated forms of the limestone dust was significantly higher than in bulk samples, but since the bulk percentage was low the enrichment factor would not have resulted in any major change to conclusions regarding the contribution of respirable rock dust to the overall airborne quartz concentration. The quartz percentage in the marble dust (untreated and treated) was very low and the respirable fractions showed no enrichment. The spectra from SEM-EDX analysis for all materials were predominantly from calcium carbonate, clay, and gypsum particles. No free quartz particles were observed. The four rock dusts used in this study are representative of those presented for use in rock dusting, but the conclusions may not be applicable to all available materials.

Endotoxin deposits on the inner surfaces of closed-face cassettes during bioaerosol sampling: a field investigation at composting facilities.

A set of 270 bioaerosol samples was taken from 15 composting facilities using polystyrene closed-face filter cassettes (CFCs). The objective was to measure the quantity of endotoxin deposits on the inner surfaces of the cassettes (sometimes referred to as 'wall deposits'). The results show that endotoxins are deposited on the inner surfaces of the CFCs through sampling and/or handling of samples. The quantity of endotoxins measured on inner surfaces range between 0.05 (the limit of detection of the method) and 3100 endotoxin units per cassette. The deposits can represent a large and variable percentage of the endotoxins sampled. More than a third of the samples presented a percentage of inner surface deposits >40% of the total quantity of endotoxins collected (filter + inner surfaces). Omitting these inner surface deposits in the analytical process lead to measurement errors relative to sampling all particles entering the CFC sampler, corresponding to a developing consensus on matching the inhalable particulate sampling convention. The result would be underestimated exposures and could affect the decision as to whether or not a result is acceptable in comparison to airborne concentration limits defined in terms of the inhalability convention. The results of this study suggest including the endotoxins deposited on the inner surfaces of CFCs during analysis. Further researches are necessary to investigate endotoxin deposits on the inner cassette surfaces in other working sectors.

Evaluation of the dark-medium objective lens in counting asbestos fibers by phase-contrast microscopy.

A Japanese round-robin study revealed that analysts who used a dark-medium (DM) objective lens reported higher fiber counts from American Industrial Hygiene Association (AIHA) Proficiency Analytical Testing (PAT) chrysotile samples than those using a standard objective lens, but the cause of this difference was not investigated at that time. The purpose of this study is to determine any major source of this difference by performing two sets of round-robin studies. For the first round-robin study, 15 AIHA PAT samples (five each of chrysotile and amosite generated by water-suspended method, and five chrysotile generated by aerosolization method) were prepared with relocatable cover slips and examined by nine laboratories. A second round-robin study was then performed with six chrysotile field sample slides by six out of nine laboratories who participated in the first round-robin study. In addition, two phase-shift test slides to check analysts' visibility and an eight-form diatom test plate to compare resolution between the two objectives were examined. For the AIHA PAT chrysotile reference slides, use of the DM objective resulted in consistently higher fiber counts (1.45 times for all data) than the standard objective (P-value < 0.05), regardless of the filter generation (water-suspension or aerosol) method. For the AIHA PAT amosite reference and chrysotile field sample slides, the fiber counts between the two objectives were not significantly different. No statistically significant differences were observed in the visibility of blocks of the test slides between the two objectives. Also, the DM and standard objectives showed no pattern of differences in viewing the fine lines and/or dots of each species images on the eight-form diatom test plate. Among various potential factors that might affect the analysts' performance of fiber counts, this study supports the greater contrast caused by the different phase plate absorptions as the main cause of high counts for the AIHA PAT chrysotile slides using the DM objective. The comparison of fiber count ratios (DM/standard) between the AIHA PAT chrysotile samples and chrysotile field samples indicates that there is a fraction of fibers in the PAT samples approaching the theoretical limit of visibility of the phase-contrast microscope with 3-degree phase-shift. These fibers become more clearly visible through the greater contrast from the phase plate absorption of the DM objective. However, as such fibers are not present in field samples, no difference in counts between the two objectives was observed in this study. The DM objective, therefore, could be allowed for routine fiber counting as it will maintain continuity with risk assessments based on earlier phase-contrast microscopy fiber counts from field samples. Published standard methods would need to be modified to allow a higher aperture specification for the objective.