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Little is known about the health effects of school-related indoor dampness and microbial exposures. In this study, we investigated dampness and dampness-related agents in both homes and schools and their association with allergy and respiratory health effects in 330 Danish pupils. Classroom dampness was identified based on technical inspection and bedroom dampness on parents' self-report. Classroom and bedroom dust was analyzed for seven microbial components. Skin prick testing determined atopic sensitization. Lung function was expressed as z-scores for forced expiratory volume in one-second (zFEV 1 ), forced vital capacity (zFVC) and the ratio zFEV 1 /zFVC using GLI-2012 prediction equations. The parents reported children's allergies, airway symptoms, and doctor-diagnosed asthma. High classroom dampness, but not bedroom dampness, was negatively associated with zFEV 1 (β-coef. -0.71; 95% CI -1.17 to -0.23) and zFVC (β-coef. -0.52; 95% CI -0.98 to -0.06) and positively with wheezing (OR 8.09; 95% CI 1.49 to 43.97). No consistent findings were found between any individual microbial components or combination of microbial components and health outcomes. Among other indoor risk factors, environmental tobacco smoke (ETS) decreased zFEV 1 (β-coef. -0.22; 95% CI -0.42 to -0.02) and zFEV 1 /zFVCratio (β-coef. -0.26; 95% CI -0.44 to -0.07) and increased upper airway symptoms (OR 1.66; 95% CI 1.03-2.66). In conclusion, dampness in classrooms may have adverse respiratory health effects in pupils, but microbial agents responsible for this effect remain unknown., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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

Indoor exposure to microbial agents is known to influence respiratory health. Besides home exposure, exposure in schools can affect respiratory health. In this study, we measured endotoxin in settled dust in primary schools in three European countries from three different geographical regions with different climates. Our aim was to characterize endotoxin levels in primary schools and evaluate associations with potential determinants. Endotoxin levels were repeatedly assessed in 23 schools in Spain (n = 7), the Netherlands (n = 10), and Finland (n = 6) using electrostatic dustfall collectors. In total, 645 measurements were taken in 237 classrooms. Endotoxin levels differed significantly between countries; Dutch schools had the highest levels, while Finnish schools showed the lowest levels. In each country, differences in endotoxin levels were observed between schools and over the sampling periods. Estimates improved after adjustment for sampling period. Factors affecting endotoxin levels in a school differed per country. In general, endotoxin levels were higher in lower grades and in classrooms with higher occupancy. School endotoxin levels may contribute significantly to total endotoxin exposure in children and teachers. As the correlation between the repeated measurements is reasonable, single endotoxin measurements form a reasonable basis for estimating annual endotoxin levels in schools., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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 β(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 β(1-3)-glucans. Information on potential determinants was obtained through questionnaires. Concentrations of endotoxin, EPS, and β(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 β(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., (© 2012 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.)

Unlabelled: Endotoxin exposures have manifold effects on human health. The geographical variation and determinants of domestic endotoxin levels in Europe have not yet been extensively described. To investigate the geographical variation and determinants of domestic endotoxin concentrations in mattress dust in Europe using data collected in the European Community Respiratory Health Survey follow-up (ECRHS II). Endotoxin levels were measured in mattress dust from 974 ECRHS II participants from 22 study centers using an immunoassay. Information on demographic, lifestyle, and housing characteristics of the participants was obtained in face-to-face interviews. The median endotoxin concentration in mattress dust ranged from 772 endotoxin units per gram (EU/g) dust in Reykjavik, Iceland, to 4806 EU/g in Turin, Italy. High average outdoor summer temperature of study center, cat or dog keeping, a high household crowding index, and visible damp patches in the bedroom were significantly associated with a higher endotoxin concentrations in mattress dust. There is a large variability in domestic endotoxin levels across Europe. Average outdoor summer temperature of study center, which explains only 10% of the variation in domestic endotoxin level by center, is the strongest meteorological determinant. The observed variation needs to be taken into account when evaluating the health effects of endotoxin exposures in international contexts., Practical Implications: The incoherent observations of the health effects of endotoxin may be partly owing to the geographical heterogeneity of endotoxin exposure. Therefore, the observed variation should be considered in further studies. Measurements of indoor endotoxin are recommended as an indicator for the level of exposures of individual domestic environments., (© 2011 John Wiley & Sons A/S.)

Dust collection by study participants instead of fieldworkers would be a practical and cost-effective alternative in large-scale population studies estimating exposure to indoor allergens and microbial agents. We aimed to compare dust weights and biological agent levels in house dust samples taken by study participants with nylon socks, with those in samples taken by fieldworkers using the sampling nozzle of the Allergology Laboratory Copenhagen (ALK). In homes of 216 children, parents and fieldworkers collected house dust within the same year. Dust samples were analyzed for levels of allergens, endotoxin, (1-->3)-beta-D-glucans and fungal extracellular polysaccharides (EPS). Socks appeared to yield less dust from mattresses at relatively low dust amounts and more dust at high dust amounts than ALK samples. Correlations between the methods ranged from 0.47-0.64 for microbial agents and 0.64-0.87 for mite and pet allergens. Cat allergen levels were two-fold lower and endotoxin levels three-fold higher in socks than in ALK samples. Levels of allergens and microbial agents in sock samples taken by study participants are moderately to highly correlated to levels in ALK samples taken by fieldworkers. Absolute levels may differ, probably because of differences in the method rather than in the person who performed the sampling. Practical Implications Dust collection by participants is a reliable and practical option for allergen and microbial agent exposure assessment. Absolute levels of biological agents are not (always) comparable between studies using different dust collection methods, even when expressed per gram dust, because of potential differences in particle-size constitution of the collected dust.

Unlabelled: A newly developed dustfall device that collects airborne dust by sedimentation has been evaluated in Danish schools. Dust collected over 140 days was compared with airborne dust sampled during single school days and settled floor dust sampled at the end of a school day. Measurements of culturable fungi in air samples corresponded well to those from the dustfall collector in mechanically but not naturally ventilated rooms. Levels of beta(1 --> 3)glucan and culturable fungi in dust from the dustfall collector and settled floor dust did not agree well. Up to 10 times more beta(1 --> 3)glucan per milligram dust was found in the dustfall collectors than in settled floor dust. Only in dust from the dustfall collector was the mass concentration of culturable fungi (cfu/mg) significantly correlated with the degree of dampness. The culturability of fungi in the 140-day dustfall samples tended to be lower than in other types of samples. Nonetheless, the results indicated that the dustfall collector can be used to estimate the median concentration of airborne culturable fungi, provided identical sampling periods are used for all samples. The collector is cost effective and simple to use., Practical Implications: Traditional ways of assessing exposure to microorganisms by sampling airborne dust or dust from floors have several disadvantages. In the present paper, a new sampler, the dustfall collector, which collects airborne dust by sedimentation over a period of months, is introduced. Tests of this collector indicate that it can be used to estimate the mean concentration of airborne culturable fungi but not beta(1 --> 3)glucan over a long period and that the method can differentiate between buildings with different levels of water damage.

Unlabelled: An environmental survey was conducted in two hospital buildings in Montana, one of which had historical water incursion on the top floors and higher prevalence of reported respiratory symptoms that improved when the occupants were away from work. We measured culturable fungi and bacteria, fungal spores, endotoxin, and sub-micron particles in air; and culturable fungi and bacteria, endotoxin, markers of fungi (extra-cellular polysaccharides specific for Penicillium/Aspergillus, ergosterol, and beta(1-->3) glucans) and cat allergen in chair and floor dusts. For the analytes measured in air, the correlation coefficients ranged from 0.43 to 0.78 (P < 0.05). In chair dust, beta(1-->3) glucan concentrations correlated with culturable fungi and ergosterol concentrations. We found that sub-micron particles and markers of microbiological agents, but not culturable microbiological agents, were significantly positively associated with the building that had both historical water damage and higher prevalence of reported respiratory symptoms. Chair dust measurements tended to be higher in the non-complaint building. These results suggest that air and floor dust measurements of marker compounds may be better indicators of current health risk in a water-damaged environment than chair dust measurements or measurements of culturable fungi or bacteria in air or settled dust., Practical Implications: Detection and quantification of nonculture-based microbiological markers and/or agents of disease may be useful methods to assess microbial contamination and to more accurately evaluate microbial exposures in the indoor environment for exposure-response studies.

Unlabelled: Vacuum cleaners with a central exhaust may lead to lower airborne dust and allergen concentrations compared with regular vacuum cleaners. In 12 houses, equipped with a central vacuum cleaning system, a standardized vacuum cleaning protocol was performed comparing airborne dust and allergen concentrations between the central system and a regular cleaner. Airborne dust concentrations during (104 vs. 95 microg/m(3)) and after vacuum cleaning (38 vs. 36 microg/m(3)) were not significantly different. In houses with a cat, Fel d 1 concentrations were also not significantly different either during (6.12 vs.5.91 ng/m(3)) and after (0.64 vs. 0.91 ng/m(3)) use of the vacuum cleaners. Airborne dust and Fel d 1 concentrations increased considerably during use of both vacuum cleaners. Compared with central vacuum cleaning systems, conventional modern vacuum cleaners do not cause higher airborne dust or cat allergen concentrations during and after use., Practical Implications: This study shows that the amount of dust or cat allergen in the air during and after vacuum cleaning is similar when comparing a central vacuum cleaner with a regular vacuum cleaner. Therefore, allergen avoidance advice given to reduce exposure of allergic asthmatic patients to allergens, does not necessarily have to include switching to a central vacuum cleaner.

Endotoxin in house dust has been shown to be associated with asthma severity. Little is known about the influence of housing characteristics on endotoxin distribution. Using standardized methods, dust was sampled from a 1m(2) site and the whole accessible carpet area in selected Wellington, New Zealand homes (n = 77). Endotoxin was measured using a Limulus Amoebocyte Lysate assay. Relative humidity and temperature were recorded using sensors placed in carpet bases. Questionnaires were used to collect information on housing characteristics. All analyses were performed for endotoxin units (EU)/mg and EU/m2 for each site. Geometric mean endotoxin levels were 22.7 EU/mg [geometric standard deviation (GSD) = 2.4] or 30,544 EU/m2 (GSD = 3.2) from the 1m(2) site, and 28.4 EU/mg (GSD = 3.4) or 5653 EU/m2 (GSD = 6.4) from the whole room. After controlling for confounding, endotoxin was positively associated with dogs inside [geometric mean ratio (GMR): 0.9-2.0], total household occupants (GMR: 1.7-2.0, for 1 m2 sample only), vacuum cleaners <1-year old (GMR: 2.3-2.7), reusing vacuum dust collection bags (GMR: 1.4-3.1), steamcleaning or shampooing the carpet (GMR: 1.4-2.2) and high relative humidity (GMR: 1.4-1.6). Lower endotoxin was associated with floor insulation (GMR: 0.4-0.8), and north-facing living rooms (GMR: 0.4-0.8). This study has identified home characteristics that could be modified to reduce endotoxin exposure.

Fungal exposure inside homes has been associated with adverse respiratory symptoms in children and adults. While fungal assessment has traditionally relied upon questionnaires, fungal growth on culture plates and spore counts, new immunoassays for extracellular polysaccharides (EPS) and beta (1-->3)-glucans have enabled quantitation of fungal agents in house dust in a more timely and cost-effective manner, possibly providing a better measure of fungal exposure. We investigated associations among measurements of EPS, beta (1-->3)-glucans and culturable fungi obtained from 23 Dutch homes. From each home, dust samples were vacuumed from the living room floor twice during the Fall, Winter and Spring seasons for a total of six collections (every 6 weeks from October 1997 to May 1998). Samples were sieved and fine dust was analyzed for EPS from Aspergillus and Penicillium spp. combined, beta (1-->3)-glucans and culturable fungi. EPS was positively associated with glucan; an increase from the 25th to the 75th percentile of glucan concentration was associated with a 1.6-fold increase in EPS concentration (95% CI = 1.3 to 2.0; p < 0.01). The most significant variables associated with EPS and glucan concentrations were the surface type that was vacuumed and the concentration of total culturable fungi (in colony forming units (CFU)/g dust), with an increase in CFU/g from the 25th to the 75th percentile associated with a 1.3 (1.1-1.6)-fold increase in glucan and a 1.7 (1.3-2.2)-fold increase in EPS concentrations. In addition, the within-home variation of EPS levels were smaller than those between homes (25,646 U/g vs. 50,635 U/g), whereas the variation of glucan levels was similar within and between homes (1,300 vs. 1,205 micrograms/g). These positive associations suggest that house dust concentrations of beta (1-->3)-glucan, and particularly those of EPS, are good markers for the overall levels of fungal concentrations in floor dust which is a surrogate for estimating airborne fungal exposure.