A cross sectional study on airborne inhalable microorganisms, endotoxin, and particles in pigeon coops - Risk assessment of exposure.

Pigeon breeding is associated with symptoms of the airways. The aim of this study is to illuminate the bacteriological and toxicological characteristics of airborne dust in pigeon coops. Airborne dust was sampled in 31 urban pigeon coops with homing and fancy pigeons, and following the dust was characterized. In total 141 different bacterial species were identified using MALDI-TOF MS, and of these 11 species are classified in risk group 2. Of the cultivable bacteria, Staphylococcus equorum was present in the highest concentration. Microorganisms in the dust were able to form biofilm, and the amount correlated positively with the number of bacteria. Next generation sequencing showed 180 genera with Acinetobacter in highest reads. On average 999 ± 225 ZOTUs were observed per sample with a Shannon-Wiener biodiversity index of 6.17 ± 0.24. Of the identified species the following have previously been suggested as causative agents of extrinsic allergic alveolitis: Alcaligenes faecalis, Bacillus subtilis, Pantoea agglomerans, Sphingobacterium spiritivorum, Thermoactinomyces sp., and Streptomyces albus. Staphylococcus was present on particles with sizes between 1.1 and > 7.0 µm with a geometric mean diameter of particles on 4.7 ± 1.1 µm. Concentrations of airborne endotoxin and dust were elevated compared to references, and the geometric mean concentrations were 102 EU/m3 and 1.07 mg dust/m3, respectively. Upon exposure to the airborne dust human granulocytes produced Reactive Oxidative Species during the first 5 min, and then no further reaction was observed. The concentrations of bacteria in general, Staphylococcus spp., and endotoxin and biodiversity were associated significantly with season, temperature and/or relative humidity, but not with type or density of pigeons. The bacterial composition and biodiversity indices were not affected by type of pigeon. In conclusion, the exposure to bacteria and endotoxin in pigeon houses should not be neglected in the evaluation of causative agents of airways symptoms among pigeon breeders.

A cohort study of cucumber greenhouse workers' exposure to microorganisms as measured using NGS and MALDI-TOF MS and biomarkers of systemic inflammation.

Work in greenhouses entails exposure to airborne fungi and bacteria. The aims of this study are to obtain knowledge about whether exposure to fungal and bacterial genera and species during work in a cucumber greenhouse is affected by work tasks, and whether a cohort of greenhouse workers' serum levels of serum amyloid A (SAA) and C-reactive protein (CRP), biomarkers of systemic inflammation, are associated with this. Data on personal exposure to airborne fungal and bacterial species measured over 4 years as well as serum levels of SAA and CRP sampled over two years were analyzed. For data analysis, the main work tasks were grouped into three different groups, called 'grouped work task'. Microorganisms were identified using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF MS) and next-generation sequencing (NGS). The 'daily exposure' of greenhouse workers' were as follows: 4.8 × 104 CFU bacteria/m3, 1.4 × 106 CFU fungi/m3, and 392 EU/m3 of endotoxin. Workers were exposed to many different microbial species including several species within the genera Acinetobacter, Bacillus, Microbacterium, Pseudomonas, Staphylococcus, and Streptomyces. The genera Ralstonia and Cladosporium were found in most samples. The exposure levels as well as the microbial composition were associated significantly with grouped work task and season with high exposures during tasks in close contact with mature and old plants and in the autumn. CRP and SAA levels were also associated with exposure level and grouped work tasks. The Shannon-Wiener indices were not different in the 3 'grouped work tasks'. Several specific species including e.g. Halomonas elongata, Stenotrophomonas maltophilia, Podosphaera fusca, and Wallemia spp. were found frequently or in high concentrations in the exposures associated with the highest levels of CRP and SAA. The microorganisms S. maltophilia, P. fusca, and Wallemia spp. were also found on the cucumber plant leaves. In conclusion, both exposure level and the species composition seem to have an effect on the serum levels of CRP and SAA of exposed workers. The greenhouse workers were exposed to only a few species characterized as human pathogens.

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.

Airborne MRSA and Total Staphylococcus aureus as Associated With Particles of Different Sizes on Pig Farms.

Airborne methicillin-resistant Staphylococcus aureus (MRSA) have previously been found on pig farms, which may lead to nasal deposition of MRSA in humans via inhalation. The anterior nares are the main niche for S. aureus, and S. aureus can cause, e.g. wound infection and pneumonia. The aim of this study was to acquire knowledge about the potential deposition of airborne MRSA, specifically, and of total S. aureus (including both methicillin-sensitive S. aureus and MRSA, in the following called S. aureus) in the different parts of the airways during occupancy on pig farms. Measurements of airborne MRSA and S. aureus were performed on four pig farms using a six and a three-stage sampler during different work tasks, such as high-pressure cleaning and everyday inspection. MRSA were quantified using MRSA-selective agar, and S. aureus were quantified using Staphylococcus selective agar. The identity of the bacteria were confirmed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The geometric mean (GM) concentrations of MRSA and S. aureus were 447 cfu/m3 air and 1.8 × 103 cfu/m3 air, respectively. The highest concentrations of MRSA and S. aureus were found among pigs in a weaner stable and during high-pressure cleaning of an empty stable, respectively. The lowest concentrations of MRSA and S. aureus were found in a stable with sick pigs and in feed-storages, respectively. Most MRSA and S. aureus were associated with particles between 7 and 12 µm. On average, the particle size fractions potentially depositing in the upper airways constituted 70%, in the primary and secondary bronchi 22%, and in the terminal bronchi and alveoli 8% of the inhalable MRSA and S. aureus concentration. Across the sampled areas, the geometric mean diameter (Dg) of particles with MRSA and S. aureus were 7.2 and 6.4 µm, respectively, and no significant difference was found between these Dgs. The Dg of the airborne particles with the studied bacterium was significantly associated with the different locations on the farms. The largest Dgs were found in the air samples from the aisles and on the fence to the pens, while the smallest Dgs were found in samples from the pens among the pigs and in samples taken at greater distances from the pigs: in the hallway, feed-storage, and entry room. In conclusion, airborne MRSA and S. aureus were found in sample fractions potentially depositing in all six parts of the airways. However, the majority was found to potentially deposit in the upper airways. The concentration of airborne MRSA and S. aureus and MRSA, as well as the fraction potentially depositing in the different parts of the airways, depended on the specific work task being performed and the location on the farm.

Generation and Characterization of Indoor Fungal Aerosols for Inhalation Studies.

In the indoor environment, people are exposed to several fungal species. Evident dampness is associated with increased respiratory symptoms. To examine the immune responses associated with fungal exposure, mice are often exposed to a single species grown on an agar medium. The aim of this study was to develop an inhalation exposure system to be able to examine responses in mice exposed to mixed fungal species aerosolized from fungus-infested building materials. Indoor airborne fungi were sampled and cultivated on gypsum boards. Aerosols were characterized and compared with aerosols in homes. Aerosols containing 10(7)CFU of fungi/m(3)air were generated repeatedly from fungus-infested gypsum boards in a mouse exposure chamber. Aerosols contained Aspergillus nidulans,Aspergillus niger, Aspergillus ustus, Aspergillus versicolor,Chaetomium globosum,Cladosporium herbarum,Penicillium brevicompactum,Penicillium camemberti,Penicillium chrysogenum,Penicillium commune,Penicillium glabrum,Penicillium olsonii,Penicillium rugulosum,Stachybotrys chartarum, and Wallemia sebi They were all among the most abundant airborne species identified in 28 homes. Nine species from gypsum boards and 11 species in the homes are associated with water damage. Most fungi were present as single spores, but chains and clusters of different species and fragments were also present. The variation in exposure level during the 60 min of aerosol generation was similar to the variation measured in homes. Through aerosolization of fungi from the indoor environment, cultured on gypsum boards, it was possible to generate realistic aerosols in terms of species composition, concentration, and particle sizes. The inhalation-exposure system can be used to study responses to indoor fungi associated with water damage and the importance of fungal species composition.

Microbial diversity in bioaerosol samples causing ODTS compared to reference bioaerosol samples as measured using Illumina sequencing and MALDI-TOF.

The importance of the microbial diversity of bioaerosols in relation to occupational exposure and work related health symptoms is not known. The aim of this paper is to gain knowledge on the bacterial and fungal communities in dust causing organic dust toxic syndrome (ODTS) and in reference dust not causing ODTS. Bacterial and fungal communities were described in personal exposure samples from grass seed workers developing ODTS, in dust generated from grass seeds causing ODTS and in dust generated from reference seeds not causing ODTS. Amplicon sequencing of the bacterial 16S rRNA gene and the fungal ITS region, as well as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) were used for identification of fungi and bacteria in personal exposure samples and in dust samples from grass seeds causing ODTS and in dust from reference grass seeds. Furthermore, activities of enzymes were measured in the same samples. The sequencing data revealed more than 150 bacterial and 25 fungal genera present in each sample. Streptomyces spp., Aspergillus fumigatus and Rhizopus microsporus were dominating in the dust causing ODTS but not in the reference dust. The dustiness in terms of Mucor sp. and R. microsporus were 100-1000 times higher for problematic seeds compared to reference seeds. The bacterial species in the dust causing ODTS included pathogenic species such as Klebsiella pneumonia and Streptomyces pneumonia, and it contained increased concentrations of total protein, serine protease, chitinase, and ß-glucosidase. Twenty-three bacterial genera covered more than 50% of the total reads in the personal and problematic seed dust. These 23 genera accounted for less than 7% of the total reads in the reference seed dust. The microbial community of the dust from the problematic seeds showed great similarities to that from the personal air samples from the workers. In conclusion, we have shown for the first time a shift in the microbial community in aerosol samples that caused ODTS compared to the reference samples that did not cause the ODTS. Furthermore, elevated enzyme activities were found in the dust causing ODTS.

Dust, endotoxin, fungi, and bacteria exposure as determined by work task, season, and type of plant in a flower greenhouse.

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.

Exposure and preventive measure to reduce high and daily exposure to Bacillus thuringiensis in potted plant production.

The bacterium Bacillus thuringiensis (Bt) is the active organism in a variety of commercially available products used worldwide as biopesticides. Bt products are applied in large outdoor areas as well as in indoor environments. Even though it has been sold for decades, not much is known about the occupational exposure to Bt. The aim of this study was to obtain knowledge about the exposure to Bt subspecies israelensis (Bti) in a propagation section in a greenhouse, where Bti is applied hourly by a spray boom, and to test a preventive measure to reduce the exposure to airborne Bti. Furthermore, we wanted to study the exposure during work with potted plants treated earlier with Bti. Exposure to aerosols with Bti was measured repeatedly by personal and stationary samplers before and after the intervention. Bti was identified by polymerase chain reaction in air and soil samples. Personal exposure to inhalable Bti in the propagation section was 3×10(5) cfu m(-3) (median level, n = 22); the personal exposure of people working with plants treated earlier with Bti was 3200 cfu m(-3) (median level, n = 17). The highest single measure was found for the person working with the spray boom (7×10(5) cfu m(-3)) but airborne Bti was present at all sampling stations in the propagation section. Bti constituted a high share of the airborne cultivable bacteria and a smaller share of the soilborne bacteria in the propagation section. In a human cell assay, spiking an aerosol sample with a product with Bti increased the inflammatory potential of an aerosol sample from the greenhouse significantly. Based on the inflammatory potential and the high personal exposure, a cover around the spray boom was built as an attempt to reduce the daily exposure to Bti. The cover reduced the personal exposure to Bti from 3.0×10(5) cfu m(-3) to 1.8×10(4) cfu m(-3). The exposure was thus reduced by a factor 17, which is a considerable reduction. Bti was present in different particle size fractions with the majority, both before and after the intervention, in the fraction of airborne particles with an aerodynamic diameter between 1.2 and 3.0 µm. The measured occupational exposure to Bti is discussed in relation to risk evaluation.

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.

Organic dust toxic syndrome at a grass seed plant caused by exposure to high concentrations of bioaerosols.

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, ß-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.