Fungal and bacterial species on biowaste workers' hands and inhalation zone, and potential airway deposition.

This study aims to investigate the microbiological working environment of biowaste workers, focusing on airborne fungal and bacterial species exposure, size distribution, and species on workers' hands. The research, conducted across six plants with 45 personal exposure assessments, revealed a total of 150 bacterial species and 47 fungal species on workers' hands, including 19 and 9 species classified in risk class 2 (RC2), respectively. Workers' exposure analysis identified 172 bacterial and 32 fungal species, with several in RC2. In work areas, 55 anaerobic bacterial species belonging to RC2 were found. Different species compositions were observed in various particle size fractions, with the highest species richness for anaerobic bacteria in the fraction potentially depositing in the secondary bronchi and for fungi in the pharynx fraction. The geometric mean aerodynamic diameter (DG) of RC2 anaerobic bacteria was 3.9 µm, <1.6 µm for Streptomyces, 3.4 µm for Aspergillus, and 2.0 µm for Penicillium. Overlapping species were identified on workers' hands, in their exposure, and in work areas, with Bacillus amyloliquefaciens, Leuconostoc mesenteroides, Bacillus cereus, Enterococcus casseliflavus, and Aspergillus niger consistently present. While the majority of RC2 bacterial species lacked documented associations with occupational health problems, certain bacteria and fungi, including Bacillus cereus, Escherichia coli, Enterobacter, Klebsiella pneumonia, Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, Lichtheimia corymbifera, Lichtheimia ramosa, and Paecilomyces variotii, have previously been linked to occupational health issues. In conclusion, biowaste workers were exposed to a wide range of microorganisms including RC2 species which would deposit in different parts of the airways.

Work in nursing homes and occupational exposure to endotoxin and bacterial and fungal species.

Indoor microbial exposure may cause negative health effects. Only little is known about the occupational microbial exposure in nursing homes and the factors that influence the exposure. The exposure in nursing homes may be increased due to close contact with elderly persons who may carry infectious or antimicrobial-resistant microorganisms and due to handling of laundry, such as used clothing and bed linen. We investigated the microbial exposure in 5 nursing homes in Denmark, by use of personal bioaerosol samples from different groups of staff members taken during a typical working day, stationary bioaerosol measurements taken during various work tasks, sedimented dust samples, environmental surface swabs, and swabs from staff members' hands. From the samples, we explored bacterial and fungal concentrations and species composition, endotoxin levels, and antimicrobial resistance in Aspergillus fumigatus isolates. Microbial concentrations from personal exposure samples differed among professions, and geometric means (GM) were 2,159 cfu/m3 (84 to 1.5 × 105) for bacteria incubated on nutrient agar, 1,745 cfu/m3 (82 to 2.0 × 104) for bacteria cultivated on a Staphylococcus selective agar, and 16 cfu/m3 air for potential pathogenic fungi incubated at 37 °C (below detection limit to 257). Bacterial exposures were elevated during bed making. On surfaces, the highest bacterial concentrations were found on bed railings. The majority of bacterial species found were related to the human skin microflora, such as different Staphylococcus and Corynebacterium species. Endotoxin levels ranged from 0.02 to 59.0 EU/m3, with a GM of 1.5 EU/m3. Of 40 tested A. fumigatus isolates, we found one multiresistant isolate, which was resistant towards both itraconazole and voriconazole, and one isolate resistant towards amphotericin B. In conclusion, we give an overview of the general microbial exposure in nursing homes and show that microbial exposures are higher for staff with more care and nursing tasks compared with administrative staff.

Health symptoms, inflammation, and bioaerosol exposure in workers at biowaste pretreatment plants.

Biowaste pretreatment plants have been built within the last years in Denmark in order to recycle pre-sorted biowaste from houses, restaurants, and industry. We investigated the association between exposure and health at six biowaste pretreatment plants (visited twice) across Denmark. We measured the personal bioaerosol exposure, took blood samples, and administered a questionnaire. Thirty-one persons participated, 17 of them twice, resulting in 45 bioaerosol samples, 40 blood samples, and questionnaire answers from 21 persons. We measured exposure to bacteria, fungi, dust, and endotoxin, the total inflammatory potential of the exposures, and serum levels of the inflammatory markers serum amyloid A (SAA), high sensitivity C-reactive protein (hsCRP), and human club cell protein (CC16). Higher exposures to fungi and endotoxin were found for workers with tasks inside the production area compared to workers with main tasks in the office area. A positive association was found between the concentration of anaerobic bacteria and hsCRP and SAA, whereas bacteria and endotoxin were inversely associated with hsCRP and SAA. A positive association between hsCRP and the fungal species Penicillium digitatum and P. camemberti were found, whereas an inverse association between hsCRP and Aspergillus niger and P. italicum were found. Staff with tasks inside the production area reported more symptoms of the nose than those working in the office area. To conclude, our results indicate that workers with tasks inside the production area are exposed to elevated levels of bioaerosols, and that this may affect workers' health negatively.

Exposure levels of dust, endotoxin, and microorganisms in the Danish recycling industry.

INTRODUCTION: Recycling of domestic waste and a number of employees in the recycling industry is expected to increase. This study aims to quantify current exposure levels of inhalable dust, endotoxin, and microorganisms and to identify determinants of exposure among recycling workers. METHODS: This cross-sectional study included 170 full-shift measurements from 88 production workers and 14 administrative workers from 12 recycling companies in Denmark. The companies recycle domestic waste (sorting, shredding, and extracting materials from waste). We collected inhalable dust with personal samplers that were analysed for endotoxin (n = 170) and microorganisms (n = 101). Exposure levels of inhalable dust, endotoxin, and microorganisms and potential determinants of exposure were explored by mixed-effects models. RESULTS: The production workers were 7-fold or higher exposed to inhalable dust, endotoxin, bacteria, and fungi than the administrative workers. Among production workers recycling domestic waste, the geometric mean exposure level was 0.6 mg/m3 for inhalable dust, 10.7 endotoxin unit (EU)/m3 for endotoxin, 1.6 × 104 colony forming units (CFU)/m³ of bacteria, 4.4 × 104 CFU/m³ of fungi (25 °C), and 1.0 × 103 CFU/m³ of fungi (37 °C). Workers handling paper or cardboard had higher exposure levels than workers handling other waste fractions. The temperature did not affect exposure levels, although there was a tendency toward increased exposure to bacteria and fungi with higher temperatures. For inhalable dust and endotoxin, exposure levels during outdoor work were low compared to indoor work. For bacteria and fungi, indoor ventilation decreased exposure. The work task, waste fraction, temperature, location, mechanical ventilation, and the company size explained around half of the variance of levels of inhalable dust, endotoxin, bacteria, and fungi. CONCLUSION: The production workers of the Danish recycling industry participating in this study had higher exposure levels of inhalable dust, endotoxin, bacteria, and fungi than the administrative workers. Exposure levels of inhalable dust and endotoxin among recycling workers in Denmark were generally below established or suggested occupational exposure limits (OEL). However, 43% to 58% of the individual measurements of bacteria and fungi were above the suggested OEL. The waste fraction was the most influential determinant for exposure, and the highest exposure levels were seen during handling paper or cardboard. Future studies should examine the relationship between exposure levels and health effects among workers recycling domestic waste.

Airborne bacterial species in indoor air and association with physical factors.

The aim of this study is to obtain knowledge about which cultivable bacterial species are present in indoor air in homes, and whether the concentration and diversity of airborne bacteria are associated with different factors. Measurements have been performed for one whole year inside different rooms in five homes and once in 52 homes. Within homes, a room-to-room variation for concentrations of airborne bacteria was found, but an overlap in bacterial species was found across rooms. Eleven species were found very commonly and included: Acinetobacter lowffii, Bacillus megaterium, B. pumilus, Kocuria carniphila, K. palustris, K. rhizophila, Micrococcus flavus, M. luteus, Moraxella osloensis and Paracoccus yeei. The concentrations of Gram-negative bacteria in general and the species P. yeei were significantly associated with the season with the highest concentrations in spring. The concentrations of P. yeei, K. rhizophila and B. pumilus were associated positively with relative humidity (RH), and concentrations of K. rhizophila were associated negatively with temperature and air change rate (ACR). Micrococcus flavus concentrations were associated negatively with ACR. Overall, this study identified species which are commonly present in indoor air in homes, and that the concentrations of some species were associated with the factors: season, ACR and RH.

A cohort study of wastewater treatment plant workers: Association between levels of biomarkers of systemic inflammation and exposure to bacteria and fungi, and endotoxin as measured using two methods.

Work in wastewater treatment plants (WWTPs) can be associated with exposure to airborne microorganisms and endotoxin from the working environment. The aim of this study was to obtain knowledge about whether serum levels of the markers of systemic inflammation, C-reactive protein (CRP) and serum amyloid A (SAA), are associated with personal exposure to endotoxin, measured using the Limulus (endotoxinLimulus) and the rFC (endotoxinrFC) assays, as well as bacteria and fungi in a cohort of WWTP workers. Exposure and blood samples were collected for 11 workers over one year. Exposure to endotoxinLimulus-day and endotoxinrFC-day correlated significantly (r = 0.80, p<0.0001, n = 104), but endotoxinLimulus-day was 4.4 (Geometric mean (GM) value) times higher than endotoxinrFC-day (p<0.0001). The endotoxinLimulus-day, endotoxinrFC-day, bacteria, and fungal exposure as well as serum levels of CRP-day (GM=1.4 mg/l) and SAA-day (GM=12 mg/l) differed between workers. Serum levels of SAAday correlated significantly with CRPday (r = 0.30, p = 0.0068). The serum levels of CRPday were associated significantly with exposure to endotoxinLimulus-day. Exposure, SAA and CRP data were also analyzed as av. of each season, and SAAseason was associated positively and significantly with endotoxinLimulus-season and endotoxinrFC-season and negatively with fungalseason exposure. In conclusion, CRPday was associated with the endotoxinLimulus-day and SAAseason with endotoxinLimulus-season and endotoxinrFC-season exposure. Thus, we hereby document that WWTP workers are exposed to airborne endotoxin which seems to have a negative impact on their health.

Airborne methicillin-resistant Staphylococcus aureus, other bacteria, fungi, endotoxin, and dust in a pigeon exhibition.

Pigeon breeding is associated with exposure to airborne microorganisms and endotoxin and with symptoms of the airways. Antibiotic resistance is a threat to human health. Some pigeons participate in national and international indoor exhibitions. This study aims to obtain knowledge about the potential human exposure to dust, endotoxin, fungi, and bacteria including the methicillin-resistant Staphylococcus aureus (MRSA) in a pigeon exhibition in Denmark. In walking areas for visitors, airborne microorganisms in different size fractions able to enter the airways were sampled and following identified. The average concentrations were: 5000 cfu fungi/m3, 1.8 × 104 cfu bacteria/m3, 37 endotoxin units/m3, and 0.18 mg dust/m3 air with the highest concentrations in-between rows with pigeon cages. The fungal species Wallemia sp. and Aspergillus versicolor and the bacterial species S. equorum and S. aureus were found in high concentrations. MRSA spa type t034 described to be associated with livestock was found in the air. Most of the S. aureus was present in the size fraction of 1.1-2.1 µm, which are particles able to enter the human terminal bronchi. In conclusion, fungi, bacteria, and endotoxin, respectively, were found in concentrations 10, 2000, and 200 times higher than outdoor references. The airborne bacteria in the exhibition were mainly species found previously in pigeon coops showing that the pigeons are the sources of exposure. The presence of airborne MRSA in the pigeon exhibition highlights the importance of also considering this environment as a potential place of exchange of resistant bacteria between animals and between animals and humans.

Work clothes as a vector for microorganisms: Accumulation, transport, and resuspension of microorganisms as demonstrated for waste collection workers.

Work clothes may act as a vector for the transport of microorganisms leading to second-hand exposure; however, this has not been studied in work environments. We investigated whether microorganisms accumulate on workers' clothes in environments with elevated microbial exposures, and whether they are transported with the clothes and subsequently resuspended to the air. To study this, we selected waste collection workers and potential transport of bacteria and fungi to waste truck cabs via clothes, and compared the microbial communities within truck cabs, in waste collection workers' personal exposure, and on clean T-shirts worn by the workers. Microbial communities were also investigated for the presence of potentially harmful microorganisms. Results showed that microorganisms accumulated in large quantities (GM = 3.69 × 105 CFU/m2/h for bacteria, GM = 8.29 × 104 CFU/m2/h for fungi) on workers' clothes. The concentrations and species composition of airborne fungi in the truck cabs correlated significantly with the accumulation and composition of fungi on clothes and correlated to concentrations (a trend) and species composition of their personal exposures. The same patterns were not found for bacteria, indicating that work clothes to a lesser degree act as a vector for bacteria under waste collection workers' working conditions compared to fungi. Several pathogenic or allergenic microorganisms were present, e.g.: Klebsiella oxytoca, K. pneumoniae, Proteus mirabilis, Providencia rettgeri, Pseudomonas aeruginosa, and Aspergillus fumigatus, A. glaucus, A. nidulans, A. niger, and various Penicillium species. The potential 'take-home' exposure to these microorganisms are of most concern for immunocompromised or atopic individuals or people with open wounds or cuts. In conclusion, the large accumulation of microorganisms on workers' clothes combined with the overlap between fungal species for the different sample types, and the presence of pathogenic and allergenic microorganisms forms the basis for encouragement of good clothing hygiene during and post working hours.

Accumulation of microorganisms on work clothes of workers collecting different types of waste - A feasibility study.

Electrostatic dust cloths have previously been used to study microorganisms in settled dust by placing the cloths horizontally on surfaces (called Electrostatic Dust Collectors, EDC). In this study, we investigate whether the same cloths, henceforth called 'E-Cloths', can be used to study accumulation of microorganisms and endotoxin on workers' clothes. This was studied as current methods have limitations. It was examined for waste collection workers, as their work environment is associated with elevated exposure to microorganisms and endotoxin. Each worker received a kit with a T-shirt with an attached E-Cloth on the front, a instruction letter, and a questionnaire. Workers wore the T-shirts during the next two workdays. Unaffected by waste type collected, it was possible to measure the accumulation of bacteria, fungi, and endotoxin from the work environment on the E-Cloths. Geometric mean concentration of 9 × 106 CFU bacteria/m2, 1 × 107 CFU fungi/m2, and 4 × 104 endotoxin units/m2 were found. In total, 100 different bacterial and 25 fungal species were found. The genus Bacillus (with 18 species) and Brevibacterium aurantiacum were among the dominating bacteria. For fungi, Penicillium brevicompactum, P. commune, Penicillium italicum, and Aspergillus niger were most often found. Importantly, mainly environmental bacteria and fungi had accumulated on the E-Cloths and only few skin-related bacterial species were present, showing that accumulation had happened from the work exposure and not workers' skin. In conclusion, the T-shirts with an E-Cloth can be used as a self-administered method for measurement of accumulation of microorganisms and endotoxin from the work environment on waste collection workers' clothes.

A characterization of bioaerosols in biowaste pretreatment plants in relation to occupational health.

Occupational exposure to microorganisms can be associated with adverse health outcomes. In this study, we assessed exposure to bioaerosols in two biowaste pretreatment plants in Denmark, which differed in location (city or countryside) and how they were built ('closed-off processes' or 'open processes'). Bioaerosol exposures were characterized by microbial concentrations in personal, stationary, sedimented dust, and hand samples, and their size distribution was assessed. Furthermore, species were identified by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF MS), and inhalable dust, endotoxin, biofilm production, the total inflammatory potential, and fungicide resistance to four fungicides (amphotericin B, caspofungin acetate, itraconazole, voriconazole) were determined. Bacterial and fungal concentrations were on average (GM) in the order of 104 cfu/m3, but ranged from 102 to 108 cfu/m3. Several species which may cause health problems were identified. Personal endotoxin exposures were on average 28 EU/m3, but both personal and stationary samples ranged from 0.6 to 2035 EU/m3. Bioaerosols had the potential to form biofilms and to induce inflammation as measured in a human cell line. Exposures were higher in the plants that outdoor reference values. Higher exposures were found in the 'open process' plant, such as in microbial concentrations, species richness, endotoxin, biofilm production, and the total inflammatory potential. Six out of 28 tested Aspergillus fumigatus isolates were resistant to fungicides (amphotericin B and voriconazole). In conclusion, there is a high exposure to bioaerosols during work in biowaste pretreatment plants, however, results also suggests that how the plant is built and functions may affect the exposures.

Wastewater treatment plant workers' exposure and methods for risk evaluation of their exposure.

Work in wastewater treatment plants (WWTPs) can be associated with respiratory symptoms and diarrhea. The aim of this study was to obtain knowledge about WWTP workers' exposure to airborne bacteria and endotoxin, and the inflammatory potential (TIP) of their exposure, and to evaluate the risk posed by the exposure by 1) calculating a hazard index and relating the exposure to suggested occupational exposure limits (OELs), 2) estimating the potential deposition of bacteria in the airways, 3) relating it to the risk group classification of bacteria by the European Union, and 4) estimating the TIP of the personal exposure. A cohort of 14 workers were followed over one year. Bioaerosols were collected using personal and stationary samplers in a grid chamber house and an aeration tank area. Airborne bacteria were identified using (MALDI-TOF MS), and TIP of exposure was measured using HL-60 cells. A significant effect of season, work task, and person was found on the personal exposure. A hazard index based on exposure levels indicates that the risk caused by inhalation is low. In relation to suggested OELs, 14% and 34% of the personal exposure were exceeded for endotoxin (≥50 EU/m3) and bacteria (≥500 CFU/m3). At least 70% of the airborne bacteria in the grid chamber house and the aeration tank area could potentially deposit in the lower respiratory tract. From the personal samples, three of 131 bacterial species, Enterobacter cloacae, Staphylococcus aureus, and Yersinia enterocolitica are classified within Risk Group 2. Seven additional bacteria from the stationary samples belong to Risk Group 2. The bacterial species composition was affected significantly by season (p = 0.014) and by sampling type/area (p = 0.001). The TIP of WWTP workers' exposure was higher than of a reference sample, and the highest TIP was measured in autumn. TIP of personal exposure correlated with bacterial exposure. Based on the geometric average exposures to endotoxin (9.2 EU/m3) and bacteria (299 CFU/m3) and based on the calculated hazard index, the risk associated with exposure is low. However, since 43 of 106 exposure levels exceed suggested OELs, the TIP of exposure was elevated and associated with bacterial exposure, and WWTP workers were exposed to pathogenic bacteria, a continued focus on preventive measures is important. The identification of bacteria to species level in personal samples was necessary in the risk assessment, and measurement of the microbial composition made the source tracking possible.

Strong variance in the inflammatory and cytotoxic potentials of Penicillium and Aspergillus species from cleaning workers' exposure in nursing homes.

Penicillium and Aspergillus are among the dominant genera of fungi in many environments. Exposure to these fungi may cause inflammation-related health effects, however the knowledge about this at species level is limited. The aim of this study was to obtain knowledge about cleaning workers' exposure to fungi and to investigate the total inflammatory potential (TIP) and the cytotoxic potential of fungal species. The fungi were obtained from the personal exposure of cleaning workers' in five nursing homes. In total 271 fungal isolates were identified using MALDI-TOF MS. The TIP and cytotoxic potential were determined for 30 different fungal isolates covering 17 species in an in vitro assay by exposing HL-60 cells to the fungal spores of each isolate. The geometric mean exposure of the cleaning workers was 351 CFU fungi/m3 air. We showed that the TIP and cytotoxicity varied among both species and isolates. At the two lowest doses, there was a positive relationship between spore concentration and TIP. The species with highest TIPs were A. candidus and P. italicum, while the most cytotoxic ones were A. niger and A. fumigatus. There was no obvious relationship between the TIP of an isolate and its cytotoxicity. The results of this study provide a better understanding of the inflammatory potential and cytotoxicity of different environmental fungal species and contribute to the risk evaluation of exposure to different Penicillium and Aspergillus species.

Towards a risk evaluation of workers' exposure to handborne and airborne microbial species as exemplified with waste collection workers.

Bioaerosol exposure is associated with health problems. The aim of this study is to evaluate whether it is possible to assess the risks posed by waste collection workers' exposure through identification and characterization of bacterial and fungal species, to which the workers are exposed. Using MALDI-TOF MS, microorganisms in waste collection workers' exposure through air, hand, and contact with the steering wheel were identified. Fungi found in high concentrations from the workers' exposure were characterized for the total inflammatory potential (TIP), cytotoxicity, and biofilm-forming capacity. In total, 180 different bacterial and 37 different fungal species in the workers' exposure samples were identified. Some of them belong to Risk Group 2, e.g. Escherichia coli, Klebsiella oxytoca, Staphylococcus aureus, and Aspergillus fumigatus, some have been associated with occupational health problems e.g. Penicillium citrinum and P. glabrum and some are described as emerging pathogens e.g. Aureobasidium pullulans. The TIP of fungal species was dose-dependent. High TIP values were found for Penicillium italicum, P. brevicompactum, P. citrinum, and P. glabrum. Several species were cytotoxic, e.g. A. niger and P. expansum, while some, e.g. P. chrysogenum, did not affect the cell viability. Based on waste workers' average inhalation rate, they inhaled up to 2.3 × 104 cfu of A. niger, 7.4 × 104 cfu of P. expansum, and 4.0 × 106 cfu of P. italicum per work day. Some species e.g. A. niger and P. citrinum were able to form biofilm. In conclusion, the workers were exposed to several species of microorganisms of which some to varying degrees can be evaluated concerning risk. Thus, some microorganisms belong to Risk Group 2, and some are described as causing agents of occupational health problems, emerging pathogens, or intrinsically antibiotic resistant. For some other species very little is known. The TIP, cytotoxicity, and ability to form biofilm of the dominating fungi support and expand previous findings. These parameters depended on the species and the dose, thus highlighting the importance of species identification and exposure level in the risk assessment of exposure.

Measures to reduce the exposure of waste collection workers to handborne and airborne microorganisms and inflammogenic dust.

Waste collection is associated with various health symptoms. The aims of this study were to obtain knowledge about exposure to bacteria, fungi, and endotoxin during waste collection, and to study whether it is possible to reduce the exposures and the total inflammatory potential (TIP) of those exposures through simple interventions. The study was performed with an initial baseline exposure assessment, a second assessment with intervention workers only, and a third with intervention and reference workers. The waste collection workers were exposed to 7.8 × 103 cfu bacteria/m3, 1.4 × 104 cfu fungi/m3, and 92 endotoxin units/m3 (geometric mean values). The potential exposures in the truck cabs were up to 23 times higher than outdoor reference concentrations. For the intervention trucks and workers, airborne fungi in the truck cab were reduced; fungi, bacteria, and yeasts on the steering wheels were reduced; and the concentration of fungi on the workers' hands was reduced. Exposures were typically highest during collection of mixed household waste, in the summer, and for collection using trucks with low loading height. The TIP was highest for the reference group sampling mixed household waste, using trucks with low loading height, in the summer. Endotoxin, bacteria, and fungi contributed to the TIP of 42 personal exposure assessments. CONCLUSION: Motivating workers to reduce exposure through simple interventions improved hand and truck cab hygiene, but only slightly reduced personal exposure to airborne bioaerosols. Exposure can be reduced by only using trucks with high loading height.

Expanded cardboard waste sorting and occupational exposure to microbial species.

Member states of the European Union have to maximize recycling. The current, Danish cardboard recycling system can be improved by increasing the kinds of cardboard products that can be recycled to include e.g. used beverage cartons and pizza boxes (i.e. an expanded cardboard fraction (ECF)). This study aims to obtain knowledge about exposure to airborne endotoxin and microorganisms at species level at different collection frequencies of ECF, and whether an increase in waste sorted fractions means that each waste fraction is collected less frequently. Bacterial and endotoxin concentrations were associated significantly with temperature inside the waste containers and endotoxin and fungal exposures with collection frequency. The concentration of fungi was highest at the truck back and for reduced collection frequencies. The geometric mean diameters of particles with bacteria were between 3.0 and 5.2 µm and with fungi between 3.8 µm and 6.0 µm. In total, 81 and 25 different bacterial and fungal species were found at the waste receiving plant, respectively. Work with ECF caused exposures to food-related microorganisms (e.g. Arthrobacter arilaitensis and Penicillium camemberti), potential pathogens (e.g. Bacillus cereus, Salmonella sp. and P. expansum), and commensal bacteria. Bacillus cereus and Salmonella were found in the particle size fraction often being swallowed. Workers collecting EFC will be at risk of being exposed to microbial species that normally are related to residual waste. It seems to be advisable with an EFC collection frequency shorter than eight weeks. However, introduction of new waste fractions has generally been associated with reduced collection frequencies.

Concentrations of Staphylococcus species in indoor air as associated with other bacteria, season, relative humidity, air change rate, and S. aureus-positive occupants.

The aim of this study was to obtain knowledge about concentrations of Staphylococcus aureus, MRSA (methicillin-resistant S. aureus), and other Staphylococcus species in indoor air in Greater Copenhagen and about factors affecting the concentrations. The effects of season, temperature, relative humidity, air change rate (ACR), other bacterial genera, area per occupant, and presence of S. aureus-positive occupants were studied. In samples from 67 living rooms, S. hominis, S. warneri, S. epidermidis, and S. capitis were found in 13-25%; S. saprophyticus, S. cohnii, and S. pasteuri in 5-10%; and S. lugdunensis, S. haemolyticus, S. caprae, S. equorum, S. kloosii, S. pettenkoferi, S. simulans, and S. xylosus in less than 3%. Staphylococcus aureus were found in two of 67 living rooms: spa type t034 (an MRSA) was recovered from a farmhouse, while spa type t509 was found in an urban home. Two species, S. equorum and S. kloosii, were found only in the farmhouse. Staphylococcus was significantly associated with season with lowest concentration and richness in winter. Genera composition was associated with ACR with smaller fractions of Staphylococcus at higher ACR, while richness was significantly and negatively associated with area per occupant. Concentration of Staphylococcus correlated positively with the total concentration of bacteria, but negatively with the total concentration of other bacteria. The concentration of Staphylococcus was not significantly associated with concentrations of the other abundant genera Bacillus, Kocuria, and Micrococcus. In offices with S. aureus-positive occupants, airborne S. aureus was not found. In conclusion, Staphylococcus species constitute a considerable proportion of the airborne bacteria in the studied homes and offices. However, both S. aureus and MRSA had very low prevalence during all seasons. Thus, transmission of S. aureus and MRSA through the air in living rooms in Copenhagen is expected to be limited. The negative associations between ACR and the fraction Staphylococcus constituted out of total bacteria, and between area per occupant and Staphylococcus richness indicate that it might be possible to affect the presence of airborne Staphylococcus in homes.

Attempts to reduce exposure to fungi, ß-glucan, bacteria, endotoxin and dust in vegetable greenhouses and a packaging unit.

Indoor handling of large amounts of plant materials occurs in different occupational settings including greenhouses and causes exposure to bioaerosols. The bioaerosol components fungi, ß-glucan, bacteria and endotoxin are involved in different airway symptoms and health effects can be dose-dependent. Therefore, there is a persistent need to reduce exposure. The aims of this study were to identify tasks causing exposure and to evaluate preventive measures aimed at reducing exposure of greenhouse workers to bioaerosols, and to study factors affecting the exposure. We have focused on different exposure scenarios; one with high short-term exposure found during clearing of old cucumber plants; the other with long-term, mid-level exposure found during tomato picking, leaf nipping, stringing up tomato plants, and packaging of cucumbers. Clearing of non-dried cucumber plants compared with clearing of dried cucumber plants significantly reduced the exposure to dust, endotoxin, bacteria, fungal spores and ß-glucan. More endotoxin and fungi are emitted and more of the emitted particles were of respirable size if the leaves were dried. Along the cucumber packaging line, exposure levels were highly specific to each personal subtask. The subtask 'unloading of cucumbers' was the source of exposure making task ventilation or shielding of the process a possibility. Elimination of leaf debris on the floor reduced the exposure to fungi significantly. However, leaf debris on the floor did not contribute significantly to the exposure to dust, endotoxin and bacteria. Furthermore, to eliminate leaf debris, it had to be cleared away and this was associated with a higher exposure to dust and endotoxin. The age of the plants affected the exposure level to bioaerosols with higher exposures from old plants. In conclusion, different tasks and subtasks cause very different exposure levels. It is possible to reduce exposure by identifying subtasks causing the exposure and by modifying work processes, e.g., not drying out of plants.

Fungi, ß-glucan, and bacteria in nasal lavage of greenhouse workers and their relation to occupational exposure.

The nose and mouth are the first regions of the respiratory tract in contact with airborne microorganisms. Occupational exposures to airborne microorganisms are associated with inflammation and different symptoms of the airways. The purpose of this study is to investigate the relation between occupational exposure to fungi, ß-glucan, and bacteria and contents of fungi, ß-glucan, and bacteria in nasal lavage (NAL) of greenhouse workers. We also studied whether contents of microorganisms in NAL were related to gender, time of the work week, and runny nose. NAL samples (n = 135) were taken Monday morning and Thursday at noon and personal exposure to inhalable bioaerosols was measured during a working day. The content of fungi and ß-glucan in NAL of men was affected by their exposure to fungi and ß-glucan. The content of fungi, ß-glucan, and bacteria in NAL was higher Thursday at noon than Monday morning. The ratios of fungi in NAL between Thursday at noon and Monday morning were 14 (median value) for men and 3.5 for women. Gender had no effect on the exposure level but had a significant effect on the content of fungi, ß-glucan, and bacteria in NAL, with the highest contents in NAL of men. On Thursdays, the median content of fungi in NAL samples of men without runny noses was 9408 cfu per NAL sample, whereas the same content for women was 595 cfu per NAL sample. Workers with runny noses had fewer fungi in NAL than workers without runny noses. A higher content of ß-glucan per fungal spore was found in NAL than in the air. This indicates that mainly the larger fungal spores or pollen grains deposit in the nose. The difference between genders and the fact that the content of fungi in NAL was significantly affected by the exposure indicate that the two genders are affected by the same exposure level differently.

Sampling, extraction and measurement of bacteria, endotoxin, fungi and inflammatory potential of settling indoor dust.

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.