Geneviève Marchand

Measurement of Endotoxins in Bioaerosols at Workplace: A Critical Review of Literature and a Standardization Issue

Endotoxins are lipopolysaccharides found in the outer membrane of most Gram-negative bacteria and cyanobacteria. Worker exposure to endotoxins has been shown in a number of work situations and is associated with both respiratory and systemic pathologies. The lack of an occupational exposure limit is mainly due to the absence of a standard protocol at the international level for sampling and analyzing airborne endotoxins. The bibliographic review in this article takes an exhaustive look at the current knowledge on measuring airborne endotoxins. It shows that, despite several reference documents at the international level, the methods used to measure endotoxin exposure differ considerably from one laboratory to another. Standardization is necessary to reduce interlaboratory variability and, ultimately, to improve the use of interstudy data. The bibliographic review presents the current status of standardization for airborne endotoxin measurement methods in the workplace and summarizes areas for further research. This article is both a reference document for all operators wishing to use such methods and a working document to build international consensus around the measurement of airborne endotoxins.

Genomic characterization of a large outbreak of Legionella pneumophila serogroup 1 strains in Quebec City, 2012.

During the summer of 2012, a major Legionella pneumophila serogroup 1 outbreak occurred in Quebec City, Canada, which caused 182 declared cases of Legionnaires disease and included 13 fatalities. Legionella pneumophila serogroup 1 isolates from 23 patients as well as from 32 cooling towers located in the vicinity of the outbreak were recovered for analysis. In addition, 6 isolates from the 1996 Quebec City outbreak and 4 isolates from patients unrelated to both outbreaks were added to allow comparison. We characterized the isolates using pulsed-field gel electrophoresis, sequence-based typing, and whole genome sequencing. The comparison of patients-isolated strains to cooling tower isolates allowed the identification of the tower that was the source of the outbreak. Legionella pneumophila strain Quebec 2012 was identified as a ST-62 by sequence-based typing methodology. Two new Legionellaceae plasmids were found only in the epidemic strain. The LVH type IV secretion system was found in the 2012 outbreak isolates but not in the ones from the 1996 outbreak and only in half of the contemporary human isolates. The epidemic strains replicated more efficiently and were more cytotoxic to human macrophages than the environmental strains tested. At least four Icm/Dot effectors in the epidemic strains were absent in the environmental strains suggesting that some effectors could impact the intracellular replication in human macrophages. Sequence-based typing and pulsed-field gel electrophoresis combined with whole genome sequencing allowed the identification and the analysis of the causative strain including its likely environmental source.

Evaluation of Bioaerosols in a Municipal Solid Waste Recycling and Composting Plant

Abstract Both the processing of solid waste and the composting of organic matter generate airborne biological contaminants. For this type of environment, some authors have proposed Threshold Limit Values (TLV) of 10,000 Colony Forming Units per cubic meter (CFU/m3) of air for total bacteria, and 1,000 CFU/m3 of air for Gram-negative bacteria. Endotoxins produced by Gram-negative bacteria can cause fever and respiratory problems. Diarrhea and gastrointestinal problems can also be caused by Gram-negative bacteria. Other researchers have found Aspergillus fumigatus, which is considered an opportunistic pathogenic fungus, in the nose and throat of people exposed to compost. The objectives of this study were to evaluate the microbiological content of indoor and outdoor air, to detect concentrations of total and Gram-negative bacteria exceeding their proposed TLV, and to suggest corrective measures based on the technical and scientific literature. Air sampling was carried out twice in one day using Andersen sam...

Evaluation of bacterial contamination and control methods in soluble metalworking fluids.

In the United States, 1.2 million workers are exposed to metalworking fluids. During operations, aerosols are produced and airborne contaminants can be inhaled. Although biocides are used to control the bacterial content of metalworking fluids, they can create health-related problems, and their efficiency remains to be proved. The objectives of this project were (1) to verify whether rigorous cleaning according to a standard protocol could reduce microbial contamination and (2) whether the use of biocides with different spectra could reduce the bacterial population. Four similar machines producing similar components were evaluated; a specific treatment was applied to each machine. The machine used as a control (1) was thoroughly cleaned prior to sampling, (2) did not undergo any major cleaning afterward, and (3) was operated without the use of any biocide. A major cleaning is a protocol described and recommended by the fluid manufacturer and was performed on the three other machines, two of which were subsequently treated with biocides weekly. Fluid samples from the four lathes were collected weekly during a 6-month period, and total bacterial and cultivable Gram-negative bacteria were analyzed for each sample. Major cleaning of the machines (120–4) did not significantly reduce the concentration of bacteria in the cutting fluids when compared with the control machine (120–3), which had not undergone major cleaning. The concentrations of total bacteria were in the 10 6 CFU/mL range for these two lathes; however, a reduction in the total number of fluid changes was observed for this machine. Bacterial flora in the cutting fluids was significantly controlled with the use of biocides. Bacteria concentrations were in the 10 3 –10 5 CFU/mL range for the lathes with the use of biocides. Since thorough cleaning is insufficient and biocides are recognized as being responsible for some worker health problems, other avenues for controlling bacterial flora in cutting fluids should be evaluated to reduce worker exposure to their bacterial contaminants.

Documentation of the endotoxins present in the ambient air of cotton fiber textile mills in Québec

Cotton workers are recognized as being at risk of developing occupational lung diseases. Some researchers have identified endotoxins as being a potential etiologic agent for some of the respiratory problems. This study wants to document the concentration of endotoxins found in the ambient air of textile mills where cotton fibers are handled and to identify the processing steps where the highest endotoxins concentrations in the air were found and the one where the relative limit values (RLVs) are exceeded. The 4 mills studied process cotton fibers. All the air samples were analyzed using the chromogenic Limulus Amoebocytes lysate LAL method using a kinetic detection principle based on the IRSSTs standard method. In this study, a large variability in the concentrations of endotoxins in the air was observed, depending on the mill, the processing step, and the time. Despite these variations, some processes can be identified as being major generators of endotoxins in the ambient air of the mills. The highest concentrations were measured in the weaving and drawing processes and reached 10,000 EU m(-3) of air. The opening, cleaning, carding, spinning and drawing processes are the other major endotoxins generating processes with concentrations from 24 to 8,700 EU m(-3) of air. The endotoxins concentrations exceeded the RLVs for 55% of the workstations in this project. This study demonstrated that endotoxins levels in the cotton industry are high and appropriate control measures are needed.

A next generation sequencing approach with a suitable bioinformatics workflow to study fungal diversity in bioaerosols released from two different types of composting plants

Composting is used all over the world to transform different types of organic matter through the actions of complex microbial communities. Moving and handling composting material may lead to the emission of high concentrations of bioaerosols. High exposure levels are associated with adverse health effects among compost industry workers. Fungal spores are suspected to play a role in many respiratory illnesses. There is a paucity of information related to the detailed fungal diversity in compost as well as in the aerosols emitted through composting activities. The aim of this study was to analyze the fungal diversity of both organic matter and aerosols present in facilities that process domestic compost and facilities that process pig carcasses. This was accomplished using a next generation sequencing approach that targets the ITS1 genomic region. Multivariate analyses revealed differences in the fungal community present in samples coming from compost treating both raw materials. Furthermore, results show that the compost type affects the fungal diversity of aerosols emitted. Although 8 classes were evenly distributed in all samples, Eurotiomycetes were more dominant in carcass compost while Sordariomycetes were dominant in domestic compost. A large diversity profile was observed in bioaerosols from both compost types showing the presence of a number of pathogenic fungi newly identified in bioaerosols emitted from composting plants. Members of the family Herpotrichiellaceae and Gymnoascaceae which have been shown to cause human diseases were detected in compost and air samples. Moreover, some fungi were identified in higher proportion in air compared to compost. This is the first study to identify a high level of fungal diversity in bioaerosols present in composting plants suggesting a potential exposure risk for workers. This study suggests the need for creating guidelines that address human exposure to bioaerosols. The implementation of technical and organizational measure should be a top priority. However, skin and respiratory protection for compost workers could be used to reduce the exposure as a second resort.

Validation of the Criteria for Initiating the Cleaning of Heating, Ventilation, and Air-Conditioning (HVAC) Ductwork Under Real Conditions

Dust accumulation in the components of heating, ventilation, and air-conditioning (HVAC) systems is a potential source of contaminants. To date, very little information is available on recognized methods for assessing dust buildup in these systems. The few existing methods are either objective in nature, involving numerical values, or subjective in nature, based on experts’ judgments. An earlier project aimed at assessing different methods of sampling dust in ducts was carried out in the laboratories of the Institut de recherche Robert-Sauvé en santé et en sécurité du travail (IRSST). This laboratory study showed that all the sampling methods were practicable, provided that a specific surface-dust cleaning initiation criterion was used for each method. However, these conclusions were reached on the basis of ideal conditions in a laboratory using a reference dust. The objective of this present study was to validate these laboratory results in the field. To this end, the laboratory sampling templates were replicated in real ducts and the three sampling methods (the IRSST method, the method of the U.S. organization National Air Duct Cleaner Association [NADCA] and that of the French organization Association pour la Prévention et l’Étude de la Contamination [ASPEC]) were used simultaneously in a statistically representative number of systems. The air return and supply ducts were also compared. Cleaning initiation criteria under real conditions were found to be 6.0 mg/100 cm2 using the IRSST method, 2.0 mg/100 cm2 using the NADCA method, and 23 mg/100 cm2 using the ASPEC method. In the laboratory study, the criteria using the same methods were 6.0 for the IRSST method, 2.0 for the NADCA method, and 3.0 for the ASPEC method. The laboratory criteria for the IRSST and NADCA methods were therefore validated in the field. The ASPEC criterion was the only one to change. The ASPEC method therefore allows for the most accurate evaluation of dust accumulation in HVAC ductwork. We therefore recommend using the latter method to objectively assess dust accumulation levels in HVAC ductwork.

The effect of the number of counted traverses on the estimation of the total spore count sampled on a non-cultivable slit impactor

Various counting rules are used for spore trap analysis. Partial count can lead to concentration errors. This paper demonstrates that the number of traverses counted affects the final results.

Bacteria emitted in ambient air during bronchoscopy—a risk to health care workers?

Background Health care workers are at risk of occupational infections, and some procedures are known to increase this risk. The aim of this study was to qualify and quantify bioaerosol concentrations during bronchoscopy to estimate the occupational risk. Methods Full-day sampling was conducted in 2 rooms while bronchoscopies were performed on patients. Two microbial air samplers were used, a wet wall cyclonic sampler and an impactor, on culture media. Identification of the culturable bacterial flora was performed with chromatographic analysis of cellular fatty acid of the isolated strain and additional biochemical tests if needed. Specific polymerase chain reaction analysis was completed on wet wall cyclonic samples for the detection of influenza A and B and Mycobacterium spp. Results A wide variety of bacteria were collected from the ambient air. All samples yielded at least 1 Staphylococcus species. Although most of the culturable bacteria identified were normal nonpathogenic flora, such as Streptococcus spp, Neisseria spp, and Corynebacterium spp, some opportunistic pathogens, such as Streptococcus pneumoniae, were found. Neither Mycobacterium spp nor influenza virus was detected with the polymerase chain reaction method during this study. Conclusions Culturable bacteria from oral, nasal, and pulmonary flora are aerosolized during bronchoscopy and could be inhaled by medical staff. The potential presence of pathogens in those aerosols could represent an occupational infection risk.

Filters from taxis air conditioning system: A tool to characterize driver's occupational exposure to bioburden?

&NA; Bioburden proliferation in filters from air conditioning systems of taxis represents a possible source of occupational exposure. The aim of this study was to determine the occurrence of fungi and bacteria in filters from the air conditioning system of taxis used for patient transportation and to assess the exposure of drivers to bioburden. Filters from the air conditioning systems of 19 taxis and 28 personal vehicles (used as controls) operating in three Portuguese cities including the capital Lisbon, were collected during the winter season. The occurrence and significance of bioburden detected in the different vehicles are reported and discussed in terms of colony‐forming units (CFU) per 1 m2 of filter area and by the identification of the most frequently detected fungal isolates based on morphology. Azole‐resistant mycobiota, fungal biomass, and molecular detection of Aspergillus species/strains were also determined. Bacterial growth was more prevalent in taxis (63.2%) than in personal vehicles (26.3%), whereas fungal growth was more prevalent in personal vehicles (53.6%) than in taxis (21.1–31.6%). Seven different azole‐resistant species were identified in this study in 42.1% taxi filters. Levels of fungal biomass were above the detection limit in 63% taxi filters and in 75% personal vehicle filters. No toxigenic species were detected by molecular analysis in the assessed filters. The results obtained show that bioburden proliferation occurs widely in filters from the air conditioning systems of taxis, including the proliferation of azole‐resistant fungal species, suggesting that filters should be replaced more frequently. The use of culture based‐methods and molecular tools combined enabled an improved risk characterization in this setting. HighlightsFilters from air conditioning system are a suitable method to characterize bioburden in vehicles.Fungi and bacteria occur widely in taxis and personal vehicles.The genera Aspergillus, Cladosporium and Penicillium were often detected.Filters should be more frequently replaced to avoid bioburden proliferation and reaerosolization.Culture based and molecular methods used in parallel enabled an improved risk characterization.