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Aims: To obtain knowledge about (i) how to sample airborne methicillin-resistant Staphylococcus aureus (MRSA) and dust in the pig farm environment including effects of sampler on (a) measured exposure, (b) MRSA survival and (c) spatial and temporal variation in exposure, and (ii) the association between exposure to MRSA, dust and optical density OD., Methods and Results: Airborne dust was sampled on five pig farms using five active and one passive samplers. Staphylococcus aureus and MRSA (as a subset of S. aureus) were quantified using selective agar media and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The Andersen sampler, electrostatic dust collectors (EDC), and Gesamtstaubprobenahme (GSP) and Institute of Occupational Medicine samplers with polycarbonate or Teflon filters were applicable for sampling airborne MRSA. The half-life of MRSA was not reduced by active sampling. A significant correlation was found between dust and S. aureus exposure within, but not between, farm section and farms. A significant spatial and temporal variation in dust and MRSA exposure was found within a stable. The dust sampling rate and the concentration of MRSA in the sampled dust decreased after 5 days of sampling., Conclusion: Sampling using the GSP can be performed for 1 h without affecting the following half-life of MRSA. Sampling for MRSA using the EDC should not exceed 3 days due to overloading and the die-off of MRSA. The measurement of OD may be used as a proxy measure for dust exposure. To obtain knowledge about potential exposure, samples should be taken repeatedly and in different areas within a stable section., Significance and Impact of the Study: Sampling method, sampling time and number of samples taken, but not force of airflow on the filter, influence the measured potential exposure to MRSA and dust., (© 2019 The Society for Applied Microbiology.)

The airborne fungal and bacterial species present in pig farm dust have not been well characterised even though these bioaerosols are known to cause inflammation and other airway maladies. In this study, the microbial species and composition in airborne dust within and between pig farms were investigated. Passively sedimenting dust from six pig farms were collected using electrostatic dust collectors. The bacterial and fungal species were identified using matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and next generation sequencing (NGS). Dust samples taken within the same stable section revealed high resemblance and stability. Constrained statistical analysis of the microbial community compositions indicated that the types of stable did not appear to have a great effect on the bacterial and fungal β-diversity. In contrast to this, the farm from which samples were taken appeared to have the greatest effect on the bacterial β-diversity, but this trend was not observed for the fungal β-diversity. The most common bacteria and fungi according to NGS data were anaerobes typically associated with the pig intestinal tract and yeasts respectively. Bacterial sedimentation varied at a rate between 10 3 and 10 9 CFU/m 2 /day, with the most common species after aerobic incubation being Aerococcus viridans and Staphylococcus equorum, while Clostridium perfringens and Staphylococcus simulans were the most common species after anaerobic incubation. A total of 28 different species of bacteria and fungi were classifiable as pathogens. In conclusion, the biodiversity in pig farm dust shows a high diversity of bacterial species. However, samples from the same stable section resembled each other, but also different sections within the same farm also resembled each other, thus indicating a high degree of community stability in the dust source. In regards to fungal identification, the biodiversity was observed to be similar between samples from different stable sections and farms, indicating a higher degree of similarities in the mycobiomes found across pig farms studied., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Dust is suspected to be an important factor in transmission of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) between pigs and pig farmers and their families. The aim of this study was to determine the rate of decay for Staphylococcus aureus and LA-MRSA in dust from swine farms. Electrostatic dust fall collectors (EDCs) were used for passive sampling of settling airborne dust in 11 stable sections from six swine farms. Extraction, plating, identification, and enumeration of cultivable S. aureus and LA-MRSA from the EDCs were performed after storage for 0-30 days postsampling. The survival of S. aureus was measured in 196 dust samples from all farms, and data were used to estimate the decay constant λ according to a model for exponential decay: N(t) = N0 × e-λt. The number of S. aureus colonies was up to 600-fold higher than the number of LA-MRSA colonies on MRSA selective agar. The data showed a good fit to the model (λ = 0.13, r2 = 0.86) even with a large difference in initial concentrations of S. aureus between stables. The loads of S. aureus and LA-MRSA in the dust were significantly reduced by storage time, and the half-life was 5 days for both S. aureus and LA-MRSA. In dust samples with high initial concentrations, LA-MRSA and S. aureus could still be cultivated 30 days after sampling. On all farms MRSA isolates belonged to the clonal complex (CC) 398, and at one farm some isolates also belonged to CC30. A screening for other Staphylococcus species in the farm dust revealed 13 different species numerically dominated by Staphylococcus equorum. Based on the exponential decay model, S. equorum had a half-life of 4 days. In conclusion, the presence of MRSA in airborne dust from five of six farms indicates that dust might be an important vehicle for transmission of LA-MRSA. LA-MRSA and S. aureus was found to survive well in farm dust with half-lives of 5 days, and dependent on the initial concentration they could be found in farm dust for weeks. The 99.9% die-off rate was 66 days for LA-MRSA. Thus, farm dust can pose an exposure risk for humans in the farm environment, but also when transported to other environments. On the other hand, the risk will decrease by time. These results provide important knowledge to diminish spread from farm environments to other environments on, e.g., tools or clothing, and in relation to cleaning of emptied LA-MRSA-positive stables., (© The Author(s) 2018. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.)

Little is known about the geographic variation and determinants of bacterial endotoxin and β-(1,3)-D-glucan in Danish house dust. In a population of 317 children, we: (i) described loads and concentrations of floor dust, endotoxin, and β-(1→3)-D-glucan and (ii) their correlations and (iii) assessed their determinants; (iv) Finally, we compared our findings with previous European studies. Bedroom floor dust was analyzed for endotoxin content by the kinetic limulus amoebocyte lysate assay and for β-(1→3)-D-glucan by the inhibition enzyme immunoassay. The parents answered questions regarding potential determinants. We found: geometric means (geometric standard deviations) 186 mg/m(2) (4.3) for dust; 5.46 × 10(3) EU/m(2) (8.0) and 31.1 × 10(3) EU/g (2.6) for endotoxin; and 142 μg/m(2) (14.3) and 0.71 × 10(3) μg/g (7.3) for β-(1→3)-D-glucan. High correlations (r > 0.75) were found between floor dust and endotoxin and β-(1→3)-D-glucan loads, while endotoxin and β-(1→3)-D-glucan concentrations were moderately correlated (r = 0.36-0.41) with the dust load. Having a carpet was positively associated with dust load and with endotoxin and β-(1→3)-D-glucan concentrations. Pet keeping, dwelling type, and dwelling location were determinants of endotoxin concentrations. No other determinants were associated with β-(1→3)-D-glucan concentrations. Compared with other European studies, we found lower β-(1→3)-D-glucan loads and concentrations but higher endotoxin loads and concentrations suggesting a geographically determined different composition of Danish floor dust compared with other European regions., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Greenhouse workers are exposed to dust, endotoxin, fungi, and bacteria potentially causing airway inflammation as well as systemic symptoms. Knowledge about determinants of exposure is a prerequisite for efficient prevention through knowledge-based reduction in exposure. The objective of this study was to assess the occupational exposure in a flower greenhouse and to investigate the impact of work tasks on the intensity and variability in exposure., Methods: Seventy-six personal full-shift exposure measurements were performed on 38 employees in a Danish flower greenhouse producing Campanula, Lavandula, Rhipsalideae, and Helleborus. The samples were gravimetrically analysed for inhalable dust. Endotoxin was assessed by the Limulus Amoebocyte Lysate test and culture-based quantification of bacteria and fungi was performed. Information on the performed tasks during sampling was extracted from the greenhouse electronic task logging system. Associations between log-transformed exposure outcomes, season, and work tasks were examined in linear mixed-effects regression with worker identity as random effect., Results: Measured concentrations ranged between 0.04 and 2.41mg m(-3) for inhalable dust and between 0.84 and 1097 EU m(-3) for endotoxin exposure, with the highest mean levels measured during Lavandula and Campanula handling, respectively. Personal exposure to fungi ranged between 1.8×10(2) and 3.4×10(6) colony-forming units (CFU) m(-3) and to bacteria between 1.6×10(1) and 4.2×10(5) CFU m(-3). Exposure to dust, endotoxin, fungi, and bacteria differed between seasons. Packing Lavandula, sticking, potting, and grading Rhipsalideae, and all examined tasks related to Campanula production except sticking increased dust exposure. Endotoxin exposure was increased during sticking Campanula and pinching or packing Rhipsalideae, and fungi exposure was elevated by subtasks performed in the research and development area for Campanula, and by potting, packing/dumping Campanula. Sticking and working with subtasks in the research and development area for Campanula increased bacteria exposure., Conclusion: This study revealed moderate dust exposure levels compared to the levels observed in other greenhouse productions and other occupations with organic dust exposure such as farming. However, high exposures to bacteria and fungi were detected during selected tasks and the proposed health-based endotoxin exposure limit of 90 EU m(-3) was exceeded in 30% of the samples, which may have health implications for the employees. Exposure levels were found to vary depending on the tasks performed, and thereby results can be used to direct task-based initiatives to reduce workplace exposures., (© The Author 2014. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.)

Selection of sampling device, sampling location and period are important first steps in the measurement of exposure to bioaerosols in indoor air. The steps following the sampling include treatment of samples and laboratory analysis. In this study, settling bacteria, endotoxin, fungi and serine protease have been measured in Danish homes using Electrostatic Dust Fall Collectors (EDCs). The effects of the presence of occupants, sampling on open surfaces versus in bookcases and treatment of samples have been studied. Concentrations of bacteria and endotoxin were significantly higher when occupants were at home than when they were absent. Across homes, higher concentrations of fungi were found in spring than in winter, as was the total inflammatory potential, while higher concentrations of protease were found in winter than in spring. The placement of the EDCs in bookcases versus on an open surface significantly affected the measured concentrations of bacteria and endotoxin. Direct extraction of EDC cloths caused a higher measured concentration of bacteria, fungi and serine protease than if EDC cloths were extracted post-storage at -20 °C. Extraction of EDC cloths caused an average of 51% and 58% extraction of bacteria and fungi respectively. In conclusion, EDCs should be placed on open surfaces during the sampling, how much occupants are present in their home during sampling and sampling season should be considered, EDC cloths should not be stored in a freezer before extraction of microorganisms, but extraction suspensions can be stored at -80 °C without affecting the number of microorganisms significantly.

Unlabelled: Indoor microbial exposure has been related to allergy and respiratory disorders. However, the lack of standardized sampling methodology is problematic when investigating dose-response relationships between exposure and health effects. In this study, different sampling methods were compared regarding their assessment of microbial exposures, including culturable fungi and bacteria, endotoxin, as well as the total inflammatory potential (TIP) of dust samples from Danish homes. The Gesamtstaubprobenahme (GSP) filter sampler and BioSampler were used for sampling of airborne dust, whereas the dust fall collector (DFC), the electrostatic dust fall collector (EDC), and vacuum cleaner were used for sampling of settled dust. The GSP assessed significantly higher microbial levels than the BioSampler, yet measurements from both samplers correlated significantly. Considerably higher levels of fungi, endotoxin, and TIP were found in the EDC compared with the DFC, and regarding fungi, the EDC correlated more strongly and significantly with vacuumed dust than the DFC. Fungi in EDC and vacuum dust correlated most strongly with airborne dust, and in particular, the measurements from the EDC associated well with those from GSP. Settled dust from the EDC was most representative of airborne dust and may thus be considered as a surrogate for the assessment of indoor airborne microbial exposure., Practical Implications: Significant discrepancies between sampling methods regarding indoor microbial exposures have been revealed. This study thus facilitates comparison between methods and may therefore be used as a frame of reference when studying the literature or when conducting further studies on indoor microbial exposure. Results also imply that the relatively simple EDC method for the collection of settled dust may be used as an alternative to otherwise tedious and time-consuming airborne dust sampling., (© 2012 John Wiley & Sons A/S.)

We describe an outbreak of sudden health problems in workers at a Danish grass seed plant after exposure to a particularly dusty lot of grass seeds. The seeds are called problematic seeds. The association between development of organic dust toxic syndrome (ODTS) and the handling of grass seeds causing exposure was assessed in a four-step model: (i) identification of exposure source, (ii) characterization of the emission of bioaerosols from the problematic and reference seeds, (iii) personal and stationary exposure measurement at the plant and (iv) repeated health examinations. The grass seeds were identified as the exposure source; the emissions of some bioaerosol components were up to 10(7) times higher from the problematic seeds than from reference seeds. Cleaning of the seeds was not enough to sufficiently reduce the high emission from the problematic seeds. Emission in terms of dust was 3.4 times as high from the problematic cleaned seeds as from cleaned reference seeds. The personal exposure reached 3 × 10(5) endotoxin units m(-3), 1 × 10(6) colony-forming units (cfu) of thermophilic actinomycetes m(-3), 8 × 10(5) cfu of Aspergillus fumigatus m(-3) and 9 × 10(6) hyphal fragments m(-3). Several workers working with the problematic seeds had symptoms consistent with ODTS. The most severe symptoms were found for the workers performing the tasks causing highest exposure. Respiratory airway protection proved efficient to avoid development of ODTS. Work with reference seeds did not cause workers to develop ODTS. Exposure was during work with the problematic seeds higher than suggested occupational exposure limits but lower than in studies where researchers for some minutes have repeated a single task expected to cause ODTS. In this study, many different bioaerosol components were measured during a whole working day. We cannot know, whether it is the combination of different bioaerosol components or a single component which is responsible for the development of ODTS. In conclusion, workers developed specific health symptoms due to the high bioaerosol exposure and were diagnosed with ODTS. Exposure to high concentrations of endotoxin, actinomycetes, fungi, hyphal fragments, β-glucan, and A. fumigatus occurred when working with a dusty lot of grass seed. Suspicion should be elicited by seeds stored without being properly dried and by seeds producing more dust than usually.

We have quantified vegetable growers' exposure to fungal bioaerosol components including (1→3)-β-d-glucan (β-glucan), total fungal spores, and culturable fungal units. Furthermore, we have evaluated factors that might affect vegetable growers' exposure to fungal bioaerosols and airborne dust. Investigated environments included greenhouses producing cucumbers and tomatoes, open fields producing cabbage, broccoli, and celery, and packing facilities. Measurements were performed at different times during the growth season and during execution of different work tasks. Bioaerosols were collected with personal and stationary filter samplers. Selected fungal species (Beauveria spp., Trichoderma spp., Penicillium olsonii, and Penicillium brevicompactum) were identified using different polymerase chain reaction-based methods and sequencing. We found that the factors (i) work task, (ii) crop, including growth stage of handled plant material, and (iii) open field versus greenhouse significantly affected the workers' exposure to bioaerosols. Packing of vegetables and working in open fields caused significantly lower exposure to bioaerosols, e.g. mesophilic fungi and dust, than harvesting in greenhouses and clearing of senescent greenhouse plants. Also removing strings in cucumber greenhouses caused a lower exposure to bioaerosols than harvest of cucumbers while removal of old plants caused the highest exposure. In general, the exposure was higher in greenhouses than in open fields. The exposures to β-glucan during harvest and clearing of senescent greenhouse plants were very high (median values ranging between 50 and 1500 ng m(-3)) compared to exposures reported from other occupational environments. In conclusion, vegetable growers' exposure to bioaerosols was related to the environment, in which they worked, the investigated work tasks, and the vegetable crop.

Background: Little is known of how the toxicity of nanoparticles is affected by the incorporation in complex matrices. We compared the toxic effects of the titanium dioxide nanoparticle UV-Titan L181 (NanoTiO2), pure or embedded in a paint matrix. We also compared the effects of the same paint with and without NanoTiO2., Methods: Mice received a single intratracheal instillation of 18, 54 and 162 μg of NanoTiO2 or 54, 162 and 486 μg of the sanding dust from paint with and without NanoTiO2. DNA damage in broncheoalveolar lavage cells and liver, lung inflammation and liver histology were evaluated 1, 3 and 28 days after intratracheal instillation. Printex 90 was included as positive control., Results: There was no additive effect of adding NanoTiO2 to paints: Therefore the toxicity of NanoTiO2 was reduced by inclusion into a paint matrix. NanoTiO2 induced inflammation in mice with severity similar to Printex 90. The inflammatory response of NanoTiO2 and Printex 90 correlated with the instilled surface area. None of the materials, except of Printex 90, induced DNA damage in lung lining fluid cells. The highest dose of NanoTiO2 caused DNA damage in hepatic tissue 1 day after intratracheal instillation. Exposure of mice to the dust from paints with and without TiO2 was not associated with hepatic histopathological changes. Exposure to NanoTiO2 or to Printex 90 caused slight histopathological changes in the liver in some of the mice at different time points., Conclusions: Pulmonary inflammation and DNA damage and hepatic histopathology were not changed in mice instilled with sanding dust from NanoTiO2 paint compared to paint without NanoTiO2. However, pure NanoTiO2 caused greater inflammation than NanoTiO2 embedded in the paint matrix.

Exposure to bioaerosols in occupational settings is associated with a range of adverse health effects. The aim of this study was to investigate the exposure levels to dust and endotoxin of people working in two cucumber nurseries and two tomato nurseries. Exposure was measured for greenhouse workers (n = 70) mainly working on harvesting cucumbers and tomatoes and clearing the plants after the harvest season. The people were exposed to between 0.2 and 15 mg inhalable dust m(-3) (median = 1.6 mg m(-3)) and between 0.5 and 400 ng inhalable endotoxin m(-3) (median = 32 ng m(-3)). The exposure to 'total dust' and endotoxin measured by stationary samplers (n = 30) in the greenhouses was low. Endotoxin was present in relatively high concentrations on cucumber leaves compared with leaves on pot plants. The Danish occupational exposure limit (OEL) for total organic dust is 3 mg m(-3) and 36% and 17% of the cucumber and tomato workers, respectively, were exposed to >3.0 mg inhalable dust m(-3). There is no OEL for endotoxin, but 'no effect levels' at approximately 15 ng m(-3) have been found. The majority of subjects (65%) were exposed to >15 ng m(-3). Significantly higher exposure was found for employees in cucumber nurseries than for employees in tomato nurseries. Clearing tomato plants after the harvest season caused a higher exposure to endotoxin than tomato harvesting. In conclusion, people working in cucumber and tomato nurseries were often exposed to high levels of inhalable dust and endotoxin. Cucumber harvest workers were exposed to significantly more dust and endotoxin than tomato harvest workers. The dust and endotoxin aerosolized during the working processes were only transported to other areas in the greenhouses to a very low degree. Cucumber and tomato leaves were identified as endotoxin reservoirs.

Objectives: Particles in ambient air are associated with such health effects as lung diseases and cancer of the lung. Exposure to bioaerosols has been found to be associated with respiratory symptoms. The toxic properties of exposure to combustion and bioaerosol particles from biofuel plants have not been studied in detail. This study investigated whether exposure to dust from biofuel plants induces DNA (deoxyribonucleic acid) damage and inflammation in exposed mice., Methods: DNA damage and inflammation were evaluated in mice exposed through the intratracheal installation of airborne dust collected at a biofuel plant at the straw storage hall and in the boiler room. The mice were given either a single dose of dust (18 or 54 microg) or four doses of 54 microg on each of four consecutive days. Control mice were exposed to a 0.9% sodium chloride solution., Results: In the mice exposed to 4 x 54 microg of dust, the lung tissue mRNA (messenger ribonucleic acid) levels of interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-2 (MIP-2) were increased more than 10-fold if the dust was from the boiler room and 30- to 60-fold if the dust came from the straw storage hall. The levels of DNA strand breaks in broncheoalveolar lavage (BAL) cells from the mice exposed to dust did not differ from those in the control samples., Conclusions: The results indicate that the instillation of dust from a biofuel plant, at doses corresponding to 2 weeks of exposure to human endotoxins, results in a strong inflammatory response without detectable DNA damage in BAL cells. The dust from the straw storage hall induced the strongest inflammatory response and had the highest concentration of most microbial components.

The aim of this study was to analyse the contribution of microbial factors to the inflammatory potency of dust (PD). Floor dust was sampled three times from 12 rooms in two schools. The potency of floor dust was measured as interleukin-8 secretion from the lung epithelial cell line A549 after exposure to dust. Measurements of endotoxin, NAGase activity and cultivable fungi in the dust were made. For endotoxin a difference was found between sampling days and for NAGase a difference was found between the schools. The carpeted staff rooms of the two schools had a significantly higher amount of dust/m(2) and endotoxin/m(2), and the PD/m(2) and NAGase activity/m(2) were also higher than in the classrooms with smooth flooring. The PD/m(2) correlated with all fungal parameters except total cultivable fungi. NAGase/m(2) correlated with PD, endotoxin and cultivable Cladosporium sp. per area, and was nearly significantly correlated to total cultivable fungi and Penicillium spp. Therefore; microbiological activity especially from fungi may contribute to the inflammatory potency of floor dust. Carpet flooring may act as a "sink" for microorganisms resulting in a higher inflammatory potency of floor dust, which may reflect building-related symptoms in occupants.

Handling of biofuels may release dust particles containing high concentrations of Hazardous microorganisms, thus representing a potential occupational health problem. We analysed the microbial dustiness of baled straw (cultivated both conventionally and ecologically) and of wood chips from piles that had been stored outdoors for up to 11 months by using total spore counting, cultivation, and measuring of endotoxin and chemical markers of fungal biomass, lipopolysaccharide, and peptidoglycan. The bacterial dustiness of straw was much greater than of wood chips whereas the fungal dustiness did not differ much. In general, samples taken from the inner part of each biofuel material were dustier than samples taken from the surface, except for fungal and bacterial biomass in wood chips and total fungi and fungal biomass in ecological straw. A considerable increase of bacterial dustiness occurred during storage over summer. Dust from ecological straw contained considerably less of bacterial components than from conventional straw and, in addition, exhibited a less pronounced increase upon storage over summer. In summary, biofuels represent sustainable energy resources of growing economic importance but may at the same time pose significant health problems. We found that storage of biofuels outdoors over summer increased the microbiological dustiness and should therefore be avoided, and that ecological straw contained less of microbe-containing dust than conventional straw and should be preferred since it reduces the exposure to harmful microbiological agents.

Exposure to organic dust originating from biofuels can cause adverse health effects. In the present study we have assessed the dustiness in terms of microbial components and particles of various biofuels by using a rotating drum as a dust generator. Microbial components from straw, wood chips, wood pellets and wood briquettes were quantified by several methods. Excellent correlations (r >/= 0.85, P < 0.0001) were found: between lipopolysaccharide (LPS) (as determined by 3-hydroxy fatty acid analysis) and endotoxin (as determined by a Limulus test), cultivable bacteria, total number of bacteria and muramic acid; between endotoxin and cultivable bacteria, total number of bacteria and muramic acid; between total number of bacteria and muramic acid; between cultivable fungi and total number of fungi. Straw was dustier than the other biofuels in terms of actinomycetes, bacteria, muramic acid, endotoxin, LPS, particle mass and number of particles. One of the wood chips studied and the straws had comparatively high dustiness in terms of fungi, while both wood pellets and wood briquettes had comparatively low dustiness in terms of all microbial components. An initially high particle generation rate of straw and wood chips decreased over time whereas the particle generation rate of wood briquettes and wood pellets increased during a 5 min rotation period. Particles of non-microbial origin may be the determining factor for the health risk in handling briquettes and pellets. Straw dust contained significantly more microorganisms per particle than did wood chip dust, probably because bacteria were most abundant in straw dust. The concentrations of endotoxin and fungi were high in wood and straw dust; dust from one of the straws contained 3610 EU/mg and dust from one of the chips contained 7.3 x 10(6) fungal spores/mg. An exposure to 3 mg of straw or wood chips dust/m(3) (the Swedish and Danish OEL of unspecific inhalable dust) could cause exposures to endotoxin and fungi higher than levels were health symptoms are seen to develop. The very different levels of dustiness in terms of particles and microbial components of different biofuels shows that dustiness is an important health-relevant factor to consider when choosing among biofuels and when designing worksites for handling of biofuels.