SARS-CoV-2 incidence in secondary schools; the role of national and school-initiated COVID-19 measures.

INTRODUCTION: Our aim was to gain insight into the effect of COVID-19 measures on SARS-CoV-2 incidence in secondary schools and the association with classroom CO2 concentration and airborne contamination. METHODS: Between October 2020-June 2021, 18 schools weekly reported SARS-CoV-2 incidence and completed surveys on school-initiated COVID-19 measures (e.g. improving hygiene or minimizing contacts). CO2 was measured in occupied classrooms twice, and SARS-CoV-2 air contamination longitudinally using electrostatic dust collectors (EDC) and analyzed using RT-qPCR. National COVID-19 policy measures varied during pre-lockdown, lockdown and post-lockdown periods. During the entire study, schools were recommended to improve ventilation. SARS-CoV-2 incidence rate ratios (IRR) were estimated by Generalized Estimating Equation (GEE) models. RESULTS: During 18 weeks follow-up (range: 10-22) SARS-CoV-2 school-incidence decreased during national lockdown (adjusted IRR: 0.41, 95%CI: 0.21-0.80) and post-lockdown (IRR: 0.60, 0.39-0.93) compared to pre-lockdown. School-initiated COVID-19 measures had no additional effect. Pre-lockdown, IRRs per 10% increase in time CO2 exceeded 400, 550 and 800 ppm above outdoor level respectively, were 1.08 (1.00-1.16), 1.10 (1.02-1.19), and 1.08 (0.95-1.22). Post-lockdown, CO2-concentrations were considerably lower and not associated with SARS-CoV-2 incidence. No SARS-CoV-2 RNA was detected in any of the EDC samples. CONCLUSION: During a period with low SARS-CoV-2 population immunity and increased attention to ventilation, with CO2 levels most of the time below acceptable thresholds, only the national policy during and post-lockdown of reduced class-occupancy, stringent quarantine, and contact testing reduced SARS-CoV-2 incidence in Dutch secondary schools. Widespread SARS-CoV-2 air contamination could not be demonstrated in schools under the prevailing conditions during the study.

Host genotype affects endotoxin release in excreta of broilers at slaughter age.

Host genotype, early post-hatch feeding, and pre- and probiotics are factors known to modulate the gut microbiome. However, there is a knowledge gap on the effect of both chicken genotype and these dietary strategies and their interplay on fecal microbiome composition and diversity, which, in turn, can affect the release of endotoxins in the excreta of broilers. Endotoxins are a major concern as they can be harmful to both animal and human health. The main goal of the current study was to investigate whether it was possible to modulate the fecal microbiome, thereby reducing endotoxin concentrations in the excreta of broiler chickens. An experiment was carried out with a 2 × 2 × 2 factorial arrangement including the following three factors: 1) genetic strain (fast-growing Ross 308 vs. slower growing Hubbard JA757); 2) no vs. combined use of probiotics and prebiotics in the diet and drinking water; and 3) early feeding at the hatchery vs. non-early feeding. A total of 624 Ross 308 and 624 Hubbard JA757 day-old male broiler chickens were included until d 37 and d 51 of age, respectively. Broilers (N = 26 chicks/pen) were housed in a total of 48 pens, and there were six replicate pens/treatment groups. Pooled cloacal swabs (N = 10 chickens/pen) for microbiome and endotoxin analyses were collected at a target body weight (BW) of 200 g, 1 kg, and 2.5 kg. Endotoxin concentration significantly increased with age (p = 0.01). At a target BW of 2.5 kg, Ross 308 chickens produced a considerably higher amount of endotoxins (Δ = 552.5 EU/mL) than the Hubbard JA757 chickens (p < 0.01). A significant difference in the Shannon index was observed for the interaction between the use of prebiotics and probiotics, and host genotype (p = 0.02), where Ross 308 chickens with pre-/probiotics had lower diversity than Hubbard JA757 chickens with pre-/probiotics. Early feeding did not affect both the fecal microbiome and endotoxin release. Overall, the results suggest that the chicken genetic strain may be an important factor to take into account regarding fecal endotoxin release, although this needs to be further investigated under commercial conditions.

Green waste compost as potential reservoirs of Legionella in the Netherlands.

OBJECTIVES: Legionella is a bacterial species able to cause influenza-like illness (Pontiac fever) or severe pneumonia (Legionnaires disease, LD). We assessed Legionella presence and concentration in composting facilities in The Netherlands. METHODS: A total of 142 samples from 23 green waste composting facilities were screened for Legionella DNA using qPCR. RESULTS: Of 142 samples, Legionella spp. DNA was detected in 97 (68%), and the subspecies L. pneumophila and L. longbeachae in 33 (23%) and one (0.7%) samples, respectively. Legionella was observed in samples from all composting facilities. The concentration of Legionella spp. DNA ranged from 103 to 105 genomic units (GU)/gram. Compost temperature was negatively correlated with the presence (odds ratio 0.67, 95% CI 0.50-0.92 per 10 degrees increase) and concentration (geometric mean ratio 0.90, 95% CI 0.83-0.97 per 10 degrees) of Legionella spp. Average humidity in the week prior to sampling was negatively correlated with the L. pneumophila concentration (geometric mean ratio 0.73, 95% CI 0.56-0.96 per increase in 10% of humidity). DISCUSSION: This study suggests that composting facilities can be regarded as reservoirs of Legionella in The Netherlands, but additional studies should target if such facilities represent a human health risk.

Stability of individual LPS-induced ex vivo cytokine release in a whole blood assay over a five-year interval.

OBJECTIVE: In epidemiological and clinical studies, whole blood assay (WBA) has been used as a measure to characterize inter-individual differences in the cytokine response of individuals exposed to inflammatory agents, such as endotoxins. Several short-time repeatability studies have shown stable cytokine levels in individuals over periods of days, weeks or months, but little is known about the long-term stability of cytokine reactivity. METHODS: We studied cytokine response levels in LPS-stimulated whole blood in a cohort of 193 farmers and agricultural industry workers at two time points with a five-year interval. RESULTS: IL-10 and IL-1ß responses measured with a five-year time interval showed a weak positive correlation (r = 0.22 and 0.27, respectively), whereas no correlation was observed for TNFα (r = 0.06). Cytokine reactivity measured repeatedly at the same time point showed high correlations (IL-10 r = 0.80, IL-1ß r = 0.53 and TNFα r = 0.74), suggesting that the observed weak correlations over time are reflective of actual variations in cytokine reactivity over time. CONCLUSIONS: Repeatability of ex vivo cytokine reactivity showed to be differential for the measured cytokines, being more stable for IL-10 and IL-1ß than for TNFα. However, in general, repeatability of ex vivo cytokine reactivity was weak, reflecting that cytokine reactivity can mostly be explained by (short term) intra-individual (immunological) or time varying environmental factors and less by genetic or other time-invariant factors. Therefore, WBA should be regarded as a viable tool to study relationships with current health status and exposure, and only partially as a predictor for a future response.

An approach to the asthma-protective farm effect by geocoding: Good farms and better farms.

BACKGROUND: The highly consistent association of growing up on a farm with a reduced asthma risk has so far been attributed to direct farm exposure. In contrast, geographic determinants of the larger environment have never been assessed. In this study, the effects of proximity to farms and environmental variables in relation to the residential address on asthma and atopy were assessed. METHODS: Addresses of 2265 children of the Bavarian arm of the GABRIELA study were converted into geocodes. Proximity to the nearest cow farm was calculated, and environmental characteristics were derived from satellite data or terrestrial monitoring. Bacterial diversity in mattress dust samples was assessed in 501 children by sequencing of the 16S rRNA amplicons. Logistic regression models were used to calculate associations between outcomes and exposure variables. RESULTS: Asthma and atopy were inversely associated with the presence of a farm within a radius of maximum 100 m. The environmental variables greenness, tree cover, soil sealing, altitude, air pollution differed not only between farm and non-farm children but also between farm children with and without another farm nearby. The latter distinction revealed strong associations with characteristics of traditional farms including a broader diversity of microbial exposure, which mainly contributed to the protective effect on asthma. In non-farm children, the protective effect of a farm nearby was completely explained by consumption of farm milk. CONCLUSIONS: Clustering of farms within a neighborhood of 100 m is strongly associated with the protective effect on asthma and may represent a more traditional style of farming with broader microbial exposure.

The relation between modeled odor exposure from livestock farming and odor annoyance among neighboring residents.

PURPOSE: Odor annoyance is an important environmental stressor for neighboring residents of livestock farms and may affect their quality of life and health. However, little is known about the relation between odor exposure due to livestock farming and odor annoyance. Even more, the relation between odor exposure and odor annoyance is rather complicated due to variable responses among individuals to comparable exposure levels and a large number of factors (such as age, gender, education) that may affect the relation. In this study, we (1) investigated the relation between modeled odor exposure and odor annoyance; (2) investigated whether other factors can affect this relation; and (3) compared our dose-response relation to a dose-response relation established in a previous study carried out in the Netherlands, more than 10 years ago, in order to investigate changes in odor perception and appreciation over time. METHODS: We used data from 582 respondents who participated in a questionnaire survey among neighboring residents of livestock farms in the south of the Netherlands. Odor annoyance was established by two close-ended questions in a questionnaire; odor exposure was estimated using the Stacks dispersion model. RESULTS: The results of our study indicate a statistically significant and positive relation between modeled odor exposure and reported odor annoyance from livestock farming (OR 1.92; 95 % CI 1.53-2.41). Furthermore, age, asthma, education and perceived air pollution in the environment are all related to odor annoyance, although they hardly affect the relation between estimated livestock odor exposure and reported odor annoyance. We also found relatively more odor annoyance reported among neighboring residents than in a previous study conducted in the Netherlands. CONCLUSIONS: We found a strong relation between modeled odor exposure and odor annoyance. However, due to some uncertainties and small number of studies on this topic, further research and replication of results is recommended.

Healthy worker survivor analysis in an occupational cohort study of Dutch agricultural workers.

OBJECTIVES: High microbial exposures in farmers and agricultural workers are associated with less atopy. Although it has been speculated that healthy worker survival could be an explanation, this has not been studied so far. Therefore, we investigated the presence of healthy worker survival in a five-year follow-up study of an occupational cohort of Dutch farmers and agricultural industry (company) workers. METHODS: We compared baseline demographic characteristics, respiratory health, atopy and endotoxin exposure of 259 workers followed up with 124 workers lost to follow-up. Additionally, baseline health status of 31 participants who had changed to lower exposure jobs at follow-up was compared to those with similar or higher exposure jobs at follow-up. RESULTS: In general, no major healthy worker survival effect was found. Nonetheless, small differences were observed between subjects included in follow-up and those lost to follow-up. Those lost to follow-up were older, had a lower peak expiratory flow, and were less often raised on a farm. Company workers lost to follow-up with a farm childhood had more often self-reported allergy, but this was not observed for subjects with atopic sensitization or other respiratory symptoms. No differences were found for any of the studied characteristics in participants with lower exposure at follow-up compared to participants with similar or higher exposure at follow-up. CONCLUSIONS: No major healthy worker survival is present in this organic dust exposed cohort. Differences between participants lost to follow-up and participants included in follow-up with regard to health characteristics are small and unlikely to explain the previously reported inverse associations between endotoxin exposure and atopy.

Exposure levels, determinants and IgE mediated sensitization to bovine allergens among Danish farmers and non-farmers.

BACKGROUND: Bovine allergens can induce allergic airway diseases. High levels of allergens in dust from stables and homes of dairy farmers have been reported, but sparse knowledge about determinants for bovine allergen levels and associations between exposure level and sensitization is available. OBJECTIVE: To investigate levels and determinants of bovine allergen exposure among dairy, pig and mink farmers (bedroom and stable), and among former and never farmers (bedroom), and to assess the prevalence of bovine allergen sensitization in these groups. METHODS: In 2007-2008, 410 settled dust samples were collected in stables and in bedrooms using an electrostatic dust-fall collector over a 14 day period among 54 pig farmers, 27 dairy farmers, 3 mink farmers as well as 71 former and 48 never farmers in Denmark. For farmers sampling was carried out both during summer and winter. Bovine allergen levels (µg/m(2)) were measured using a sandwich ELISA. Determinants for bovine allergen exposure in stables and bedrooms were explored with mixed effect regression analyses. Skin prick test with bovine allergen was performed on 48 pig farmers, 20 dairy farmers, 54 former and 31 never farmers. RESULTS: Bovine allergen levels varied by five orders of magnitude, as expected with substantially higher levels in stables than bedrooms, especially for dairy farmers. Bovine allergen levels in bedrooms were more than one order of magnitude higher for dairy farmers compared to pig farmers. Former and never farmers had low levels of bovine allergens in their bedroom. Bovine allergen levels during summer appeared to be somewhat higher than during winter. Increased bovine allergen levels in the bedroom were associated with being a farmer or living on a farm. Mechanical ventilation in the bedroom decreased bovine allergen level, significant for dairy farmers ß=-1.4, p<0.04. No other significant effects of either sampling or residence characteristics were seen. Allergen levels in dairy stables were associated to type of dairy stable, but not to other stable or sampling characteristics. Sensitization to bovine allergens was only found in one pig farmer. CONCLUSION: This study confirms high bovine allergen levels in dairy farms, but also suggests sensitization to bovine allergens among Danish farmers to be uncommon. Furthermore the importance of a carrier home effect on allergen load is emphasized. Whether the risk for bovine sensitization is related to the allergen level in the stable or the dwelling remains to be determined.

Monitoring of occupational and environmental aeroallergens-- EAACI Position Paper. Concerted action of the EAACI IG Occupational Allergy and Aerobiology & Air Pollution.

Exposure to high molecular weight sensitizers of biological origin is an important risk factor for the development of asthma and rhinitis. Most of the causal allergens have been defined based on their reactivity with IgE antibodies, and in many cases, the molecular structure and function of the allergens have been established. Significant information on allergen levels that cause sensitization and allergic symptoms for several major environmental and occupational allergens has been reported. Monitoring of high molecular weight allergens and allergen carrier particles is an important part of the management of allergic respiratory diseases and requires standardized allergen assessment methods for occupational and environmental (indoor and outdoor) allergen exposure. The aim of this EAACI task force was to review the essential points for monitoring environmental and occupational allergen exposure including sampling strategies and methods, processing of dust samples, allergen analysis, and quantification. The paper includes a summary of different methods for sampling and allergen quantification, as well as their pros and cons for various exposure settings. Recommendations are being made for different exposure scenarios.

Shifted T-cell polarisation after agricultural dust exposure in mice and men.

RATIONALE: A low prevalence of asthma and atopy has been shown in farmers and agricultural workers. However, in these workers, a higher prevalence of respiratory symptoms has been reported, in which T helper 1 (Th1) and/or Th17 responses may play a role. AIM: We investigated the effect of exposure to dust extracts (DEs) from different farms on airway inflammation and T-cell polarisation in a mouse model and assessed T-cell polarisation in agricultural workers from the same farms. METHODS: DEs were prepared from settled dust collected at cattle and pig farms and bulb and onion industries. Mice were exposed to phosphate-buffered saline (PBS), DEs, house dust mite (HDM) or HDM+DE via nasal instillation, four times per week during 5 weeks. Hyperresponsiveness, airway inflammation, IgE levels and T-cell polarisation were assessed. Th-cell and T cytotoxic (Tc)-cell subsets were investigated in peripheral blood samples from 33 agricultural workers and 9 non-exposed controls. RESULTS: DEs induced interleukin(IL)-17, IL-1ß and IL-6 in mouse lung homogenates. DE-exposed mice had more mixed inflammatory infiltrates in the lungs, and more neutrophils compared with PBS-exposed mice. DEs protected against the HDM-induced Th2 response and methacholine hyperresponsiveness. Interestingly, occupationally exposed humans had higher frequencies of Th cells spontaneously expressing IL-17 and interferon γ compared with controls. CONCLUSION: Chronic exposure to different types of farm dust induces a Th/Tc-17 inflammatory response in mice and agricultural workers. This may contribute to the low prevalence of Th2-related diseases but may constitute a risk for other chronic respiratory diseases.

Endotoxin, extracellular polysaccharides, and ß(1-3)-glucan concentrations in dust and their determinants in four European birth cohorts: results from the HITEA project.

UNLABELLED: Early-life exposure to microbial agents may play a protective role in asthma and allergies development. Geographical differences in the prevalence of these diseases exist, but the differences in early-life indoor microbial agent levels and their determinants have been hardly studied. We aimed to describe the early-life levels of endotoxin, extracellular polysaccharides (EPS), and ß(1-3)-glucans in living room dust of four geographically spread European birth cohorts (LISA in Germany, PIAMA in the Netherlands, INMA in Spain, and LUKAS2 in Finland) and to assess their determinants. A total of 1572 dust samples from living rooms of participants were analyzed for endotoxin, Penicillium/Aspergillus EPS, and ß(1-3)-glucans. Information on potential determinants was obtained through questionnaires. Concentrations of endotoxin, EPS, and ß(1-3)-glucans were different across cohorts. Concentrations of endotoxin and EPS were respectively lower and higher in INMA than in other cohorts, while glucans were higher in LUKAS2. Season of sampling, dog ownership, dampness, and the number of people living at home were significantly associated with concentrations of at least one microbial agent, with heterogeneity of effect estimates of the determinants across cohorts. In conclusion, both early-life microbial exposure levels and exposure determinants differ across cohorts derived from diverse European countries. PRACTICAL IMPLICATIONS: This study adds evidence of variability in the levels of indoor endotoxin, extracellular polysaccharide, and ß(1-3)-glucans across four geographically spread European regions. Furthermore, we observed heterogeneity across regions in the effect of exposure determinants. We hypothesize that the variations observed in our study may play a role in the differences in asthma and allergies prevalences across countries.

Ex vivo cytokine release reflects sensitivity to occupational endotoxin exposure.

Not everyone exposed to endotoxin develops respiratory symptoms, even at very high exposure levels. The aim was to investigate whether ex vivo lipopolysaccharide (LPS)-induced cytokine release may be predictive of individual sensitivity to occupational endotoxin exposure. In 412 agricultural workers, tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-10 release was measured in supernatants from LPS-stimulated whole blood, lung function was measured, and respiratory symptoms were assessed by questionnaire. For each cytokine, the population was dichotomised into low and high responders according to median cytokine concentrations. Endotoxin exposure levels were determined based on 249 personal exposure measurements. High IL-10 responders had a higher prevalence of airway symptoms than low IL-10 responders (odds ratios between 2.03 and 5.10; p<0.05). TNF-alpha response was positively, but not significantly, associated with symptoms, whereas no relationship was found between IL-1beta response and symptoms. For all three cytokines, subjects with above-median responses showed significant positive dose-response relationships between endotoxin exposure and asthma symptoms, and significant associations between endotoxin exposure and a lower forced expiratory volume in 1 s (p<0.05). In contrast, exposure-response relationships were weak and statistically nonsignificant for low responders. The ex vivo inflammatory response to LPS reflects, to a large extent, whether individuals are susceptible to adverse respiratory effects induced by high occupational endotoxin exposure.

Different respiratory phenotypes are associated with isocyanate exposure in spray painters.

Associations have been observed between exposure to isocyanates, consisting mainly of oligomers, and respiratory symptoms and isocyanate specific sensitisation in spray painters. The aim of the present study was to assess associations between isocyanate exposure and more objective respiratory effect measures such as bronchial hyperresponsiveness (BHR), baseline spirometry and exhaled nitric oxide (eNO) in a subset of spray painters. Methacholine challenge and eNO measurements were performed in 229 workers. Questionnaires and blood samples were obtained. Specific immunoglobulin (Ig)E and IgG to hexamethylene di-isocyanate were assessed in serum using various assays. Personal exposure was estimated by combining personal task-based inhalatory exposure measurements and time-activity information. Workers with higher isocyanate exposure were more often hyperresponsive (prevalence ratio comparing the 75th versus 25th percentile of exposure 1.8). In addition, significant exposure-related decreased forced expiratory volume in one second (FEV(1)), FEV(1)/forced vital capacity ratio and flow-volume parameters independent of BHR were found. BHR was more prevalent among sensitised workers. This was statistically significant for only IgG-ImmunoCAP (Phadia, Uppsala, Sweden) positive workers. eNO was not associated with exposure although slightly elevated eNO levels in specific IgG positive subjects were found. The current study provides evidence that exposure to isocyanate oligomers is related to asthma with bronchial hyperresponsiveness as a hallmark, but also shows independent chronic obstructive respiratory effects resulting from isocyanate exposure.

Exhaled nitric oxide in endotoxin-exposed adults: effect modification by smoking and atopy.

OBJECTIVES: Occupational exposure to endotoxin is associated with non-allergic asthma and other airway inflammatory reactions. Little is known about the role of mucosal nitric oxide (NO) production in endotoxin-induced airway inflammation. The objective was to explore exposure-response relationships between occupational endotoxin exposure and fractional concentrations of exhaled NO (FE(NO)) and study the role of FE(NO) as an intermediate factor in the relationship between endotoxin exposure and asthma-like symptoms. METHODS: FE(NO) was measured online in 425 farmers and agricultural processing workers. For each participant (cumulative) endotoxin level was modelled on the basis of 249 personal measurements and job history. Atopy was assessed as specific serum IgE to common inhalant allergens, and other health data and personal characteristics by standardised questionnaires. RESULTS: A significant positive exposure-response relationship was found between endotoxin and FE(NO), but only in non-atopic, non-smoking subjects (p = 0.001). FE(NO) was significantly associated with current wheeze and other asthma-like symptoms irrespective of atopy and current smoking. Associations between endotoxin exposure and symptoms changed slightly after adjusting for FE(NO). CONCLUSIONS: A positive association was found between occupational endotoxin exposure and exhaled nitric oxide in non-smoking, non-atopic adults. Increased FE(NO) was associated with asthma-like symptoms, but the role of FE(NO) as an intermediate factor between endotoxin exposure and airway symptoms appears to be limited.

Exposure-response analysis of allergy and respiratory symptoms in endotoxin-exposed adults.

The objective of the present study was to investigate exposure-response relationships between current endotoxin exposure and allergic and respiratory symptoms in adults, taking into account farming exposures during childhood. A cross-sectional study was conducted among 877 Dutch farmers and agricultural industry workers in 2006. Based on 249 full-shift personal airborne endotoxin samples, a job-exposure matrix was constructed to assign endotoxin exposure levels to all participants. Associations between endotoxin exposure and questionnaire data on symptoms were studied by multiple logistic regression. Adjusted odds ratios (OR) for an interquartile range increase in endotoxin level were elevated for respiratory symptoms such as wheezing (OR 1.41 (95% confidence interval 1.16-1.72)), wheezing with shortness of breath (1.50 (1.18-1.90)) and daily cough (1.29 (1.03-1.62)). In contrast, endotoxin was strongly associated with a decreased prevalence of hay fever (0.62 (0.49-0.78)). Workers who had grown up on a farm had a lower prevalence of hay fever, but no evidence was found of effect modification by farm childhood. In conclusion, occupational endotoxin exposure in adulthood is associated with an increased risk of asthma-like symptoms but a reduced prevalence of hay fever.

Determinants of microbial exposure in grain farming.

OBJECTIVES: Exposure to organic dust containing high concentrations of microorganisms is common in grain farming, although the farmers have practices to counteract microbial growth to obtain optimal grain yields. We investigated the influence of weather and production practices on personal microbial exposure during grain work. METHODS: Airborne dust was collected by personal sampling during threshing and storage work on 92 Norwegian farms. The personal exposure for bacteria, endotoxin, fungal spores and hyphae, beta-(1-->3)-glucans and actinomycetes was quantified and compared with climatic data expressed as fungal forecasts from the grain growth season and production practices as reported by farmers. RESULTS: Farmers were exposed to a geometrical mean of 4.4 mg m(-3) inhalable dust [geometrical standard deviation (GSD) = 4.0], 4 x 10(6) m(-3) bacteria and fungal spores (GSD = 5.2 and 5.9, respectively), 5.9 x 10(3) EU m(-3) of endotoxins (GSD = 8.6), 2 x 10(5) m(-3) actinomycetes (GSD = 15.3), 120 mug m(-3) beta-(1-->3)-glucans (GSD = 4.7) and 5 x 10(5) AU m(-3) of hyphae (GSD = 4.4). Univariate associations were found between one or several of these microbial factors and work operation, visible fungal damage, grain species, lodging of grain, storage technology or harvester type. As assessed by general linear models, storage work was the main predictive determinant for microbial exposure, although grain species and visible fungal damage also were also important. Wet and warm weather throughout the grain growth season were associated with elevated exposure for inhalable dust, beta-(1-->3)-glucans, endotoxins and hyphae during threshing. The beta-(1-->3)-glucan exposure could biologically be explained by the fungal spore and hyphal exposure, both variables contributing equally. However, spores were most important during storage work, whereas only hyphae were predictive during threshing. CONCLUSIONS: Farmers were exposed to high levels of microorganisms and their components during dusty grain work. Dust prevention and protection may reduce microbial exposure, and may be particularly important in areas with frequent fungal forecasts, when fungal damage has been observed, during storage work or when handling barley.

Assessment of soy aeroallergen levels in different work environments.

BACKGROUND: Airborne soybean hull proteins are known causes of asthma epidemics around harbours and soy processing plants. Soy flour dust proteins may cause occupational allergy in food and feed industries. OBJECTIVE: To compare enzyme immunoassays (EIAs) for soy hull and soy flour aeroallergens, exposure assessment in various work environments. METHODS: Airborne dust samples (n=324) from soy unloading and/or processing plants, the animal feed industry and pig stables were analysed by two soy flour assays: one assay for measuring complete soy hull proteins and two assays for measuring the purified low-molecular-weight (LMW) soy hull allergens. RESULTS: Immunoblotting confirmed strong differences between antibody specificities and soy preparations. The results of the two soy flour assays and the assay for measuring complete soy hull proteins were highly correlated (r>0.85). The two LMW soy hull assays also showed a strong mutual correlation (r=0.91), but much less correlation with assays for measuring soy flour and complete soy hull. The levels of LMW soy hull proteins were the highest at sites of soybean unloading or processing, while soy flour levels were particularly high in the soy and animal feed industry. CONCLUSIONS: The optimal EIA procedure for soy aeroallergen exposure assessment depends on the type of work environment and the local soy dust composition. Thus, the type of work environment should always be taken into account in future soy allergy studies in order to prevent a possible underestimation of the workers' actual risk of developing soy allergy.

Atopy and new-onset asthma in young Danish farmers and CD14, TLR2, and TLR4 genetic polymorphisms: a nested case-control study.

BACKGROUND: Evidence exists that exposure to high levels of microbial agents such as endotoxin in the farm environment decreases the risk of atopic sensitization. Genetic variation in innate immunity genes may modulate the response to microbial agents and thus influence susceptibility to asthma and atopy. OBJECTIVE: To study potential associations between single nucleotide polymorphisms (SNPs) in CD14, Toll-like receptor 2 (TLR2), and TLR4 genes, and atopy and new-onset asthma in young farmers. METHODS: A nested case-control study was conducted within a cohort of 1901 young Danish farmers. We genotyped 100 new-onset asthma cases and 88 control subjects for three CD14 SNPs, three TLR2 SNPs, and two TLR4 SNPs. Atopy at baseline (defined as a positive skin prick test to one or more common inhalant allergens) was found in 17 asthma cases (17.0%) and in 17 controls (19.3%). RESULTS: The CD14/-260T allele was significantly associated with less atopy [odds ratio (OR) 0.39; 95% confidence interval (CI) 0.21-0.72, additive genetic model], whereas the CD14/-651T allele was positively associated with atopy (OR 2.53; 95% CI 1.33-4.80). Similar results were obtained by haplotype analysis. Stratified analysis by farm childhood showed stronger effects of both CD14 SNPs on atopy among farmers who were born and raised on a farm, although no significant interaction was found. No associations between CD14, TLR2, or TLR4 genotypes and new-onset asthma were found. CONCLUSION: The CD14/-260 and CD14/-651 promoter polymorphisms are associated with atopy prevalence among young adults exposed to farm environments.

Optimized methods for fungal alpha-amylase airborne exposure assessment in bakeries and mills.

BACKGROUND: In order to enable reproducible and comparable exposure measurements of fungal alpha-amylase (alpha-amylase) in different laboratories and countries, the entire procedure from sampling of airborne dust to measuring extracted samples (including standards and the used enzyme) immunoassays must be standardized. The aim of this study was to establish optimal elution and assay conditions. METHODS: A parallel sampler was used for simultaneous collection of 10 samples of inhalable dust in bakeries and mills in Germany, England, the Netherlands and Spain. Three enzyme-immunoassays (EIAs) for detection of fungal alpha-amylase based on monoclonal antibodies or polyclonal antibodies were used for the measurement of the parallel-sampled filters (n=432) extracted using several methods. The results were analysed by regression analysis of variance. Additional filters (n=54) were extracted and analysed using two EIAs to investigate the storage stability of the extracts. RESULTS: Although alpha-amylase concentrations correlated well (r> or =0.88), differences were found between the EIAs in the sensitivity and nominal values (up to a mean factor 5.8). The best elution medium for airborne filters (phosphate-buffered saline 'PBS' with 0.05% Tween-20) led to 1.2 to two times higher alpha-amylase allergen yields than extraction in PBS only, while higher Tween-20 concentrations decreased the extracted alpha-amylase yield. During storage of frozen dust/filter extracts for 3-4 months at -20 degrees C, a loss of approximately 40% of measurable alpha-amylase was observed, which could be partially prevented by addition of 0.1% casein to the medium directly after extraction. CONCLUSION: Although the effects of only a few of many possible causes of variation were investigated, for these factors a clear choice could be made with regard to optimal elution conditions and the use of validated EIAs with calibrated standards, thus making significant progress towards a completely standardized procedure for airborne alpha-amylase measurements.

Airborne exposure to wheat allergens: measurement by human immunoglobulin G4 and rabbit immunoglobulin G immunoassays.

BACKGROUND: Exposure to airborne wheat allergens in the bakery trade is associated with a high risk of occupational allergy and asthma. Control and reduction of allergen exposure require relatively simple but reliable monitoring techniques. We developed new rabbit IgG-based enzyme immunoassays (EIA) for wheat allergens, which might be a convenient alternative for the thus far used human IgG4 inhibition assay. METHODS: The reactivity and specificity of rabbit antibodies were assessed by EIA and immunoblotting, and compared with those of IgE from wheat-sensitized bakers, and with the antibodies used in the IgG4 inhibition EIA. An IgG inhibition and a sandwich EIA were developed for analysis of airborne dust samples. RESULTS: Human IgG4 and rabbit IgG inhibition EIAs had comparable sensitivities, with limits of detection (LOD) between 18 and 88 ng/mL, while the sandwich EIA was much more sensitive (LOD<0.2 ng/mL). Human IgG4 and rabbit IgG reacted in immunoblotting with most of the IgE-binding wheat proteins, although with quantitative differences. All three assays showed a strong reaction with wheat proteins, and some cross-reactivity with rye and barley, but were further highly specific for cereal flour proteins. Concentrations measured with the three EIAs in 432 airborne dust samples were highly correlated (r>0.95) and their absolute values showed less than 10-20% differences. CONCLUSION: The rabbit IgG EIAs are valid substitutes for the human IgG4 inhibition EIA, with important practical advantages. The inhibition EIA is recommended for routine wheat allergen measurements. The sandwich EIA may be used to measure low allergen levels, as in short task-related exposure measurements or in subfractions of airborne dust samples.