Geographical variation and the determinants of domestic endotoxin levels in mattress dust in Europe

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.     

Comparison of sampling methods for the assessment of indoor microbial exposure

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

Detection of streptomycetes in house dust--comparison of culture and PCR methods

Streptomycetes are gram-positive, spore producing, filamentous bacteria common in soil, but also present in indoor environments. They are potent producers of secondary metabolites and inducers of inflammatory responses in vitro. Polymerase chain reaction (PCR)- and culture-based detection methods for streptomycetes in house dust samples were compared. A total of 47 dust samples were investigated, and the presence of streptomycetes was determined by cultivation on tryptone-yeast-extract-glucose agar and PCR. The 16S rRNA gene of actinomycete isolates from house dust was partially sequenced to investigate if they belong to the genus Streptomyces. Both PCR and culture showed more frequent occurrence of streptomycetes in moisture-damaged homes, although the results did not correlate well. The occurrence of streptomycetes in house dust was associated with moisture damage of the home. The amount of Streptomyces-specific PCR amplification product was significantly higher in dust from moisture-damaged homes than in homes with no moisture damage (P < 0.05, Mann-Whitney U test). A correlation between streptomycetes and moisture damage, although not statistically significant, was also observed when using binary data, e.g. presence or absence of streptomycetes or moisture damage (P = 0.054 for PCR, and P = 0.127 for culture, Fisher's exact test). Altogether, the presence of streptomycetes in house dust seems to indicate the presence of moisture damage in the building.     

Household endotoxin reduction in the Louisa Environmental Intervention Project for rural childhood asthma

Endotoxin exacerbates asthma. We designed the Louisa Environmental Intervention Project (LEIP) and assessed its effectiveness in reducing household endotoxin and improving asthma symptoms in rural Iowa children. Asthmatic school children (N = 104 from 89 homes) of Louisa and Keokuk counties in Iowa (aged 5-14 years) were recruited and block-randomized to receive extensive (education + professional cleaning) or educational interventions. Environmental sampling collection and respiratory survey administration were done at baseline and during three follow-up visits. Mixed-model analyses were used to assess the effect of the intervention on endotoxin levels and asthma symptoms in the main analysis and of endotoxin reduction on asthma symptoms in exploratory analysis. In the extensive intervention group, dust endotoxin load was significantly reduced in post-intervention visits. The extensive compared with the educational intervention was associated with significantly decreased dust endotoxin load in farm homes and less frequent nighttime asthma symptoms. In exploratory analysis, dust endotoxin load reduction from baseline was associated with lower total asthma symptoms score (Odds ratio: 0.52, 95% confidence interval: 0.29-0.92). In conclusion, the LEIP intervention reduced household dust endotoxin and improved asthma symptoms. However, endotoxin reductions were not sustained post-intervention by residents.     

Characterization of endotoxin and 3-hydroxy fatty acid levels in air and settled dust from commercial aircraft cabins

Endotoxin was measured in air and dust samples collected during four commercial aircraft flights. Samples were analyzed for endotoxin biological activity using the Limulus assay. 3-hydroxy fatty acids (3-OH FA) of carbon chain lengths C10:0-C18:0 were determined in dust by gas chromatography-ion trap tandem mass spectrometry. The geometric mean (geometric standard deviation) endotoxin air level was 1.5 EU/m3 (1.9, n = 28); however, significant differences were found by flight within aircraft type. Mean endotoxin levels were significantly higher in carpet dust than in seat dust (140 +/- 81 vs. 51 +/- 25 EU/mg dust, n = 32 each, P < 0.001). Airborne endotoxin levels were not significantly related to either carpet or seat dust endotoxin levels. Mean 3-OH FA levels were significantly higher in carpet dust than in seat dust for C10:2, C12:0, and C14:0 (P < 0.001 for each), while the mean level of C16:0 was significantly higher in seat dust than in carpet dust (P < 0.01). Carpet dust endotoxin was significantly, but moderately, correlated with 3-OH-C12:0 and 3-OH-C14:0 (Pearson r = 0.52 and 0.48, respectively), while correlation of seat dust endotoxin with individual 3-OH FAs depended on the test statistic used. Mean endotoxin potency was significantly higher for carpet dust than for seat dust (6.3 +/- 3.0 vs. 3.0 +/- 1.4 EU/pmol LPS, P < 0.0001). Mean endotoxin levels in the air and dust of commercial aircraft cabins were generally higher than mean levels reported in homes and office buildings. These results suggest that exposure route and dust source are important considerations when relating endotoxin exposure to specific health outcomes.     

Levels and predictors of endotoxin in mattress dust samples from East and West German homes

East-West differences in prevalence of asthma and allergies were suggested to be associated with lifestyle factors. To describe endotoxin levels in mattress dust samples from East and West German homes collected approximately 10 years after reunification. To identify factors that may account for an East-West difference. Dust was collected from mattresses of 2157 infants and 2108 mothers living in Leipzig (former East Germany) and Munich (West Germany). Endotoxin was measured using a chromogenic kinetic Limulus amoebocyte lysate test. Data on bedding, dwelling, and housing characteristics, and occupants' behavior were collected using a self-administered questionnaire. Endotoxin levels were significantly higher in Leipzig compared with Munich for the infants' mattresses but not for the mothers' mattresses. Apart from this, predictors for the mothers' and the infants' mattresses were very similar. Pet-ownership and contact with animals outside the home were most influential. Endotoxin levels in mattress dust were highest in summer and increased with the number of persons living in the household. The overall percentage of variability in endotoxin levels explained by these factors was low. Endotoxin levels were associated with lifestyle factors discussed within the framework of the hygiene hypothesis. None of these factors explains the difference in infant's mattress dust endotoxin between Leipzig and Munich or could be used as a surrogate for endotoxin. PRACTICAL IMPLICATIONS: Endotoxin levels in mattress dust are associated with a number of factors discussed in the framework of the hygiene hypothesis; among other things they are associated with pet-ownership, contact with pets and number of persons living in the home. However, none of these factors and not even a combination of factors explains the variability of endotoxin levels between homes.     

Microbial characteristics in homes of asthmatic and non‐asthmatic adults in the ECRHS cohort

Microbial exposures in homes of asthmatic adults have been rarely investigated; specificities and implications for respiratory health are not well understood. The objectives of this study were to investigate associations of microbial levels with asthma status, asthma symptoms, bronchial hyperresponsiveness (BHR), and atopy. Mattress dust samples of 199 asthmatics and 198 control subjects from 7 European countries participating in the European Community Respiratory Health Survey II study were analyzed for fungal and bacterial cell wall components and individual taxa. We observed trends for protective associations of higher levels of mostly bacterial markers. Increased levels of muramic acid, a cell wall component predominant in Gram‐positive bacteria, tended to be inversely associated with asthma (OR's for different quartiles: II 0.71 [0.39‐1.30], III 0.44 [0.23‐0.82], and IV 0.60 [0.31‐1.18] P for trend .07) and with asthma score (P for trend .06) and with atopy (P for trend .02). These associations were more pronounced in northern Europe. This study among adults across Europe supports a potential protective effect of Gram‐positive bacteria in mattress dust and points out that this may be more pronounced in areas where microbial exposure levels are generally lower.     

Domestic exposure to endotoxin and respiratory morbidity in former smokers with COPD

Indoor air pollution has been linked to adverse chronic obstructive pulmonary disease (COPD) health, but specific causative agents have not yet been identified. We evaluated the role of indoor endotoxin exposure upon respiratory health in former smokers with COPD. Eighty‐four adults with moderate to severe COPD were followed longitudinally and indoor air and dust samples collected at baseline, 3 and 6 months. Respiratory outcomes were repeatedly assessed at each time point. The associations between endotoxin exposure in air and settled dust and health outcomes were explored using generalizing estimating equations in multivariate models accounting for confounders. Dust endotoxin concentrations in the main living area were highest in spring and lowest in fall, while airborne endotoxins remained steady across seasons. Airborne and dust endotoxin concentrations were weakly correlated with one another (r_s = +0.24, P = 0.005). Endotoxin concentrations were not significantly associated with respiratory symptoms, rescue medication use, quality of life, or severe exacerbations. In vitro whole‐blood assays of the pro‐inflammatory capacity of PM₁₀ filters with and without endotoxin depletion demonstrated that the endotoxin component of indoor air pollution was not the primary trigger for interleukin‐1β release. Our findings support that endotoxin is not the major driver in the adverse effects of indoor PM upon COPD morbidity.     

Assessment of endotoxin levels in the home and current asthma and wheeze in school-age children

The relationship between household endotoxin and asthma in children is not clear. To further investigate the relationship between sources of endotoxin and childhood asthma, we conducted a case-control study of children with and without asthma and examined their more frequent household exposures in the home. Children ages 6-13 years with current asthma (n = 70) or wheeze only (n = 19) were sex and age matched (+/-1 year) to 107 controls. Play area and mattress dust were collected for endotoxin analysis. Atopic status was determined by skin prick testing for allergies. A family size of >4 per household was associated with higher endotoxin levels (EU/mg) in the bed dust (P < 0.05). Passive smoking (P < 0.05) and the presence of a cat were associated with higher levels of endotoxin in mattress dust. Endotoxin levels in either the play dust or the bed dust did not differ between cases and controls. Within atopic cases, those with higher endotoxin loads (EU/m2) in bed or play areas were more likely to miss school for chest illness (P < 0.05). In this study, household endotoxin is not a risk factor for current asthma overall but may be associated with increased severity in children with atopic asthma. PRACTICAL IMPLICATIONS: This study did not find that household sources of endotoxin were associated with asthma. However, within atopic asthmatics, asthma severity (as measured by a history of being kept home from school because of a chest illness in the past year) was associated with higher levels of endotoxin in dust from the child's bed. There is a need to further investigate the nature of the relationship between household endotoxin and asthma severity in children which could lead to better management of childhood asthma.     

Endotoxin and β(1→3)-Glucan in House Dust and the Relation with Home Characteristics: A Pilot Study in 25 German Houses

Abstract Residential microbial exposure has been suggested to be involved in the development of asthma. This paper describes bacterial endotoxin and mold β(1→3)-glucan levels in house dust and the relationship to selected home characteristics. Dust was sampled from mattresses and living room and bedroom floors of 25 houses in Germany. Endotoxin and β(1→3)-glucan levels ranged from 200-48,600 EU/g dust (100-32,900 EU/m² sampled surface) and 182-3,507 μg/g (157-3,652 μg/m²), respectively. Bio-contaminant levels were highest on living room floors and lowest in mattresses. Dust, endotoxin and β(1→3)-glucan levels were 2–3 times higher on living room floors of centrally heated houses built after 1970 compared to older individually heated houses. This was not found for mattresses and bedroom floors. No associations between biocontaminant levels and other selected home characteristics (temperature, relative humidity, damp spots and insulation of windows) were found. β(1→3)-glucan levels were associated with total culturable fungi (per m²) in house dust, as well as with the fungal genus Alternaria (per g dust and per m²). In conclusion endotoxin and β(1→3)-glucan were readily detectable in house dust and significantly associated with heating system and/or age of the home.     

Dust sampling methods for endotoxin - an essential, but underestimated issue

Exposure to farming environment in early life has been associated with lower risk for allergic diseases possibly caused by increased exposure to endotoxin. The aims of this study were to compare the reproducibility of different sampling methods for endotoxin, and to determine whether environmental characteristics have different effect on endotoxin levels of different sample types. The reproducibility of sampling methods (bed dust, floor dust, vacuum cleaner dust bag dust, settled dust and air samples) was studied with repeated sampling (five visits during 1 year) in five farming and five urban homes. To examine determinants of endotoxin for different types of dust sample, sampling was conducted once in 12 farming and 17 urban homes. Endotoxin was analyzed using Limulus Amebocyte Lysate assay. Bed dust samples had the best reproducibility (intraclass correlation, ICC=66%), but the difference between farming and non-farming homes was not clear with this sample type. The reproducibility of floor (ICC=52%) and settled dust (ICC=51%) was moderate. With these sample types the difference between farming and non-farming homes was clear. Settled dust had some seasonal variation. Based on this study, the best compromise for sampling for endotoxin appears to be floor dust sample followed by bed and settled dust samples. Practical Implications Endotoxins have been widely measured, even though the validity of different sample types to reflect the endotoxin exposure level of an indoor environment is poorly known. This study shows that bed dust samples have the best reproducibility, but they do not reflect the differences in exposure due to environmental factors such as farming. Floor dust samples with moderate reproducibility may be the best choice for sampling of endotoxin in large field studies.     

Determinants of house dust endotoxin in three European countries - the AIRALLERG study

The comparison of endotoxin levels between study populations and countries is limited as a result of differences in sampling, extraction, and storage procedures. The objective of this study is to assess the levels and determinants of endotoxin in mattress and living room floor dust samples from three European countries, namely, Germany, the Netherlands, and Sweden, using a standardized sampling, storage, and analysis protocol. The mattress and living room floor dust was collected from the homes of 1065 German, Dutch, and Swedish (pre-)school children. All the samples were collected in the cool season and analyzed for endotoxin in a central laboratory. The determinants were assessed by a standardized questionnaire. The endotoxin concentrations in mattress and living room floor dust were found to be the highest in German homes and lowest in the Swedish ones. Differences between the geometric means were small (factor 1.1-1.7). Most of the associations between endotoxin concentrations and potential determinants were not statistically significant and heterogeneous across countries. However, keeping pets and having more than four persons living in the home were consistently associated with up to 1.7-fold higher endotoxin concentrations in mattress and floor dust. Furthermore, having carpets or rugs, and opening the windows frequently was associated with up to 3.4-fold and 1.3-fold higher endotoxin concentrations in living room floor dust, respectively. The proportion of variance explained by the questionnaire variables was generally low. In conclusion, the data on housing characteristics did not accurately predict the endotoxin concentrations in house dust, and could only partly explain the differences between countries. PRACTICAL IMPLICATIONS: The differences between the endotoxin concentrations in German, Dutch, and Swedish homes are small. House dust endotoxin concentrations are associated with a number of housing factors, such as pet-ownership, floor cover, number of persons living in the home, and ventilation. The variability of the endotoxin levels between homes and countries can only be partly explained by these factors.     

Fungal and endotoxin measurements in dust associated with respiratory symptoms in a water-damaged office building

We investigated the associations of fungal and endotoxin levels in office dust with respiratory health in 888 (67% participation) occupants of a water-damaged building. We analyzed floor and chair dusts from 338 workstations for culturable fungi and endotoxin. Based on averages, we ranked each floor of the building as low, medium, or high for occupants' exposure to each of these agents. Multivariate logistic regression models for building-related symptoms included this ranking of fungi and endotoxin, age, gender, race, smoking status, and duration of occupancy. Using floor dust measures, we found significantly increased odds for lower respiratory symptoms [wheeze, chest tightness, attacks of shortness of breath, and attacks of cough: odds ratios (OR) = 1.7 (95% confidence interval (CI): 1.02-2.77) to 2.4 (95% CI: 1.29-4.59)], throat irritation [OR = 1.7, (95% CI: 1.06-2.82)], and rash/itchy skin [OR = 3.0, (95% CI: 1.47-6.19)] in the highest fungal exposure group compared to the lowest, with generally linear exposure-response relationships. Nonlinear relationships were observed for many of these symptoms and endotoxin in floor dust. Interaction models showed that endotoxin modified effects of fungi on respiratory symptoms. Our findings of exposure interactions and exposure-response relationships of fungal and endotoxin with increased risk of building-related symptoms contribute to an understanding of the role of microbial agents in building-related asthma and respiratory and systemic symptoms. PRACTICAL IMPLICATIONS: Our demonstration of exposure-response relationships between measurements of fungi and/or endotoxin in floor dusts and building-related symptoms implies that microbial agents in floor dust may be a good surrogate measure for dampness-related bioaerosol exposure, considering that measurements of microbial agents in air often fail to demonstrate the associations between exposure and health. In addition, our finding that endotoxin exposure may change the effect of fungal exposure (and vice versa) on respiratory heath suggests that exposure to both fungi and endotoxin should be assessed in epidemiological investigations examining the effect of fungal or endotoxin exposure on respiratory health in indoor environments.     

Exposure matrices of endotoxin, (1 → 3)-β-d-glucan, fungi, and dust mite allergens in flood-affected homes of New Orleans

This study examined: (i) biocontaminant levels in flooded homes of New Orleans two years after the flooding; (ii) seasonal changes in biocontaminant levels, and (iii) correlations between biocontaminant levels obtained by different environmental monitoring methods. Endotoxin, (1 → 3)-β-d-glucan, fungal spores, and dust mite allergens were measured in 35 homes during summer and winter. A combination of dust sampling, aerosolization-based microbial source assessment, and long-term inhalable bioaerosol sampling aided in understanding exposure matrices. On average, endotoxin found in the aerosolized fraction accounted for < 2% of that measured in the floor dust, suggesting that vacuuming could overestimate inhalation exposures. In contrast, the (1 → 3)-β-d-glucan levels in the floor dust and aerosolized fractions were mostly comparable, and 25% of the homes showed aerosolizable levels even higher than the dust-borne levels. The seasonal patterns for endotoxin in dust and the aerosolizable fraction were different from those found for (1 → 3)-β-d-glucan, reflecting the temperature and humidity effects on bacterial and fungal contamination. While the concentration of airborne endotoxin followed the same seasonal trend as endotoxin aerosolized from surfaces, no significant seasonal difference was identified for the concentrations of airborne (1 → 3)-β-d-glucan and fungal spores. This was attributed to the difference in the particle size; smaller endotoxin-containing particles can remain airborne for longer time than larger fungal spores or (1 → 3)-β-d-glucan-containing particles. It is also possible that fungal aerosolization in home environments did not reach its full potential. Detectable dust mite allergens were found only in dust samples, and more commonly in occupied homes. Levels of endotoxin, (1 → 3)-β-d-glucan, and fungi in air had decreased during the two-year period following the flooding as compared to immediate measurements; however, the dust-borne endotoxin and (1 → 3)-β-d-glucan levels remained elevated. No conclusive correlations were found between the three environmental monitoring methods. The findings support the use of multiple methods when assessing exposure to microbial contaminants.     

Particle size distributions and concentrations of airborne endotoxin using novel collection methods in homes during the winter and summer seasons

A comparison study of novel collection methods for airborne bacteria and endotoxin was performed in an environmentally controlled chamber and in pilot-field studies. Airborne particulate matter was collected in swirling liquid impingers, air-monitoring filter cassettes, and with a micro-orifice uniform deposit impactor (MOUDI) to evaluate aerodynamic particle size distributions. Environmentally controlled chamber studies showed that impingers and MOUDI recovered significantly more airborne bacteria than filter cassettes, whereas collection methods for airborne endotoxin were not significantly different. In addition, total airborne bacteria and endotoxin concentrations were measured indoors and outdoors at three homes in Boulder, CO during winter and summer seasons. Indoor concentrations collected with the three different samplers were significantly different for airborne endotoxin, but not for airborne bacteria. Total airborne bacteria indoors and outdoors significantly varied with seasons. Outdoor airborne endotoxin significantly varied with season; no seasonal variation was seen for indoor airborne endotoxin. Indoor and outdoor levels were not significantly different for both airborne bacteria and endotoxin. The largest proportion of endotoxin was associated with airborne particulate matter <1 microm. PRACTICAL IMPLICATIONS: This study compared sampling methods for airborne endotoxin, a potent and nonspecific immune system stimulant which can induce negative health responses. The data from this study showed that swirling liquid impingers and the micro-orifice uniform deposit impactor (MOUDI) recovered significantly more airborne endotoxin than the more widely adapted method of collecting airborne endotoxin on membrane filters, when collection methods were applied in realistic settings (homes). The MOUDI measured the particle size distribution of airborne endotoxin, which can be useful for determining endotoxin respiratory toxicity and its health effects.     

Characterization and variability of endotoxin and 3-hydroxy fatty acids in an office building during a particle intervention study

Air and dust samples were collected on two floors of an office building during a double-blind particle intervention study to examine spatial and temporal variability of airborne endotoxin over a period of weeks, and to characterize endotoxin activity and lipopolysaccharide (LPS) content in carpet and chair dust. Air samples were collected on multiple days within and across weeks. Dust samples were collected from carpets and chairs one day per week for three weeks. Endotoxin was measured using a Limulus assay. Dust samples were analyzed for LPS by determination of 3-hydroxy fatty acids (3-OHFAs) using gas chromatography-mass spectrometry. The geometric mean (geometric standard deviation) for 96 indoor air samples was 0.24 (1.6) EU/m3. Significant within-floor spatial variation of airborne endotoxin was found (P < 0.0001, n = 80). Temporal variability of airborne endotoxin was not significant across weeks. Mean (+/- SD) endotoxin levels in carpet dust (59 +/- 9.3 EU/mg dust, n = 12) and in chair dust (38 +/- 7.7 EU/mg dust, n = 10) were significantly different (P < 0.001). Carbon chain length-dependent differences in 3-OHFA levels by dust source and floor were found. Enhanced air filtration did not significantly affect airborne endotoxin (P = 0.62); however, total dust mass and total endotoxin in carpet dust samples increased significantly after enhanced surface cleaning (P < 0.01). These findings suggest that spatial variability, dust source, and surface cleaning may influence building occupant exposures to endotoxin.     

Comparison of two dust collection methods for reservoir indoor allergens and endotoxin on carpets and mattresses

Variable methods of dust collection may lead to uncertainty in the measurement of biomarkers. The purpose of this study was to examine the effect of two different dust collection devices on dust weight, Der p 1, Fel d 1, and endotoxin levels. We compared: (1) a nylon mesh sock inserted between the furniture attachment and the vacuum hose (the reference method) and (2) the ALK device. Duplicate dust samples were collected for 2 min from 2 m(2) of 37 living room floors and from each longitudinal half of 37 mattresses. Measurement of Der p 1 and Fel d 1 were by double monoclonal antibody enzyme-linked immunosorbent assay (ELISA) and endotoxin by a Limulus Amobocyte Lysate assay. Geometric mean ratios (95% confidence intervals) were calculated to show the differences between sampling devices for each measurement. Compared with the ALK device, the reference method collected significantly more dust from floors (sevenfold) and mattresses (threefold) and more total Der p 1, Fel d 1, and endotoxin in both sites. Floor, but not mattress, Der p 1 concentrations were also significantly higher (threefold) using our reference method. We recommend that, in order to minimize sampling device bias, allergen and endotoxin are expressed as a concentration, and that the bed is considered the major source of allergen exposure. Practical Implications Dust sampling equipment can influence the dust yield. In order to have confidence in comparisons of allergen and endotoxin reservoir levels between centers, standardization in the use of sampling equipment is important.     

The mycobiomes and bacteriomes of sputum,saliva, and home dust

Respiratory microbiome is an understudied area of research compared to other microbiomes of the human body. The respiratory tract is exposed to an array of environmental pollutants, including microbes. Yet, we know very little about the relationship between environmental and respiratory microbiome. The primary aim of our study was to compare the mycobiomes and bacteriomes between three sample types from the same participants, including home dust, saliva, and sputum. Samples were collected from 40 adolescents in a longitudinal cohort. We analyzed the samples using 16s bacterial rDNA and ITS fungal rDNA gene sequencing, as well as quantitative PCR with universal fungal and bacterial primers. Results showed that home dust had the greatest alpha diversity between the three sample types for both bacteria and fungi. Dust had the highest total fungal load and the lowest total bacterial load. Sputum had greater bacterial diversity than saliva, but saliva had greater fungal diversity than sputum. The distribution of major bacterial phyla differed between all sample types. However, the distribution of major fungal classes differed only between sputum and saliva. Future research should examine the biological significance of the taxa found in each sample type based on microbial ecology and associations with health effects.     

Endotoxin levels in settled airborne dust in European schools: the HITEA school study

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.     

Determinants of house dust,endotoxin, and β‐(1→3)‐D‐glucan in homes of Danish children

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/m² (4.3) for dust; 5.46 × 10³EU/m² (8.0) and 31.1 × 10³EU/g (2.6) for endotoxin; and 142 μg/m² (14.3) and 0.71 × 10³ 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.