Seasonal Variations of Indoor Microbial Exposures and Their Relation to Temperature,Relative Humidity,and Air Exchange Rate

Indoor microbial exposure has been related to adverse pulmonary health effects. Exposure assessment is not standardized, and various factors may affect the measured exposure. The aim of this study was to investigate the seasonal variation of selected microbial exposures and their associations with temperature, relative humidity, and air exchange rates in Danish homes. Airborne inhalable dust was sampled in five Danish homes throughout the four seasons of 1 year (indoors, n = 127; outdoors, n = 37). Measurements included culturable fungi and bacteria, endotoxin, N-acetyl-beta-d-glucosaminidase, total inflammatory potential, particles (0.75 to 15 μm), temperature, relative humidity, and air exchange rates. Significant seasonal variation was found for all indoor microbial exposures, excluding endotoxin. Indoor fungi peaked in summer (median, 235 CFU/m³) and were lowest in winter (median, 26 CFU/m³). Indoor bacteria peaked in spring (median, 2,165 CFU/m³) and were lowest in summer (median, 240 CFU/m³). Concentrations of fungi were predominately higher outdoors than indoors, whereas bacteria, endotoxin, and inhalable dust concentrations were highest indoors. Bacteria and endotoxin correlated with the mass of inhalable dust and number of particles. Temperature and air exchange rates were positively associated with fungi and N-acetyl-beta-d-glucosaminidase and negatively with bacteria and the total inflammatory potential. Although temperature, relative humidity, and air exchange rates were significantly associated with several indoor microbial exposures, they could not fully explain the observed seasonal variations when tested in a mixed statistical model. In conclusion, the season significantly affects indoor microbial exposures, which are influenced by temperature, relative humidity, and air exchange rates.     

Air- and dustborne mycoflora in houses free of water damage and fungal growth

Typically, studies on indoor fungal growth in buildings focus on structures with known or suspected water damage, moisture, and/or indoor fungal growth problems. Reference information on types of culturable fungi and total fungal levels are generally not available for buildings without these problems. This study assessed 50 detached single-family homes in metropolitan Atlanta, Ga., to establish a baseline of "normal and typical" types and concentrations of airborne and dustborne fungi in urban homes which were predetermined not to have noteworthy moisture problems or indoor fungal growth. Each home was visually examined, and samples of indoor and outdoor air and of indoor settled dust were taken in winter and summer. The results showed that rankings by prevalence and abundance of the types of airborne and dustborne fungi did not differ from winter to summer, nor did these rankings differ when air samples taken indoors were compared with those taken outdoors. Water indicator fungi were essentially absent from both air and dust samples. The air and dust data sets were also examined specifically for the proportions of colonies from ecological groupings such as leaf surface fungi and soil fungi. In the analysis of dust for culturable fungal colonies, leaf surface fungi constituted a considerable portion (>20%) of the total colonies in at least 85% of the samples. Thus, replicate dust samples with less than 20% of colonies from leaf surface fungi are unlikely to be from buildings free of moisture or mold growth problems.     

Air- and Dustborne Mycoflora in Houses Free of Water Damage and Fungal Growth

Typically, studies on indoor fungal growth in buildings focus on structures with known or suspected water damage, moisture, and/or indoor fungal growth problems. Reference information on types of culturable fungi and total fungal levels are generally not available for buildings without these problems. This study assessed 50 detached single-family homes in metropolitan Atlanta, Ga., to establish a baseline of “normal and typical” types and concentrations of airborne and dustborne fungi in urban homes which were predetermined not to have noteworthy moisture problems or indoor fungal growth. Each home was visually examined, and samples of indoor and outdoor air and of indoor settled dust were taken in winter and summer. The results showed that rankings by prevalence and abundance of the types of airborne and dustborne fungi did not differ from winter to summer, nor did these rankings differ when air samples taken indoors were compared with those taken outdoors. Water indicator fungi were essentially absent from both air and dust samples. The air and dust data sets were also examined specifically for the proportions of colonies from ecological groupings such as leaf surface fungi and soil fungi. In the analysis of dust for culturable fungal colonies, leaf surface fungi constituted a considerable portion (>20%) of the total colonies in at least 85% of the samples. Thus, replicate dust samples with less than 20% of colonies from leaf surface fungi are unlikely to be from buildings free of moisture or mold growth problems.     

Fungal and Bacterial Communities in Indoor Dust Follow Different Environmental Determinants

People spend most of their time inside buildings and the indoor microbiome is a major part of our everyday environment. It affects humans’ wellbeing and therefore its composition is important for use in inferring human health impacts. It is still not well understood how environmental conditions affect indoor microbial communities. Existing studies have mostly focussed on the local (e.g., building units) or continental scale and rarely on the regional scale, e.g. a specific metropolitan area. Therefore, we wanted to identify key environmental determinants for the house dust microbiome from an existing collection of spatially (area of Munich, Germany) and temporally (301 days) distributed samples and to determine changes in the community as a function of time. To that end, dust samples that had been collected once from the living room floors of 286 individual households, were profiled for fungal and bacterial community variation and diversity using microbial fingerprinting techniques. The profiles were tested for their association with occupant behaviour, building characteristics, outdoor pollution, vegetation, and urbanization. Our results showed that more environmental and particularly outdoor factors (vegetation, urbanization, airborne particulate matter) affected the community composition of indoor fungi than of bacteria. The passage of time affected fungi and, surprisingly, also strongly affected bacteria. We inferred that fungal communities in indoor dust changed semi-annually, whereas bacterial communities paralleled outdoor plant phenological periods. These differences in temporal dynamics cannot be fully explained and should be further investigated in future studies on indoor microbiomes.     

The incidence of fungal spores in the ambient air and inside homes: Evidence from London

Little research has been carried out in London concerning fungal spore prevalence yet this information may help to elucidate geographical patterns of asthma and hay fever. Although many types of spore reach peak concentrations outdoors in late-summer, the incidences in the indoor environment may be more important through the winter because of heating and poor ventilation. Daily average concentrations of fungal spores in the ambient atmosphere were monitored with a Burkard volumetric spore trap on an exposed roof in North London from autumn 1991 until the summer of 1992. Indoor spore measurements were taken in 19 homes in the vicinity through the winter months, both by direct air sampling using a portable Burkard sampler and by dust culture. Trends in the occurrence and concentrations of fungal spores indoors and outdoors were examined. Relationships between the abundance of selected allergenic fungi and features of the houses were analysed including age of dwelling, dampness, cleanliness and presence of pets.Aspergillus andPenicillium were the most frequently occurring spore types in the homes. Overall, high spore incidence was associated with dampness and dust accumulation. The outdoor spore samples revealed generally low concentrations through the winter until March when concentrations of many types includingCladosporium, Epicoccum andAlternaria increased in abundance in response to the warmer weather. Even during the late-spring and early-summer, concentrations of most fungal spores were notably below those reported for rural sites.     

Fungi isolated from asthmatic homes in the Taipei area.

Indoor fungi in Taipei were surveyed during 1969-1974 using Sabouraud's agar plates. No remarkable yearly variation of fungal incidence was observed. The fungal population was found to be different from home to home. The dominant members were Aspergillus, Penicillium, Mycelia sterila, Oospora, Rhodotorula, Hormodendrum and yeasts. The order of frequency of fungi found indoors was quite different from that of outdoor fungi. The fungal spores were more numerous during summer months, and this trend was more apparent for Aspergillus, Oospora, Rhodotorula, and Hormodendrum. The kind and the incidence of fungi found in asthmatic homes were not different from those detected in non-asthmatic homes, except that in asthmatic homes there were greater numbers of fungi.     

Microbial community composition but not diversity changes along succession in arctic sand dunes

The generality of increasing diversity of fungi and bacteria across arctic sand dune succession was tested. Microbial communities were examined by high‐throughput sequencing of 16S rRNA genes (bacteria) and internal transcribed spacer (ITS) regions (fungi). We studied four microbial compartments (inside leaf, inside root, rhizosphere and bulk soil) and characterized microbes associated with a single plant species (Deschampsia flexuosa) across two sand dune successional stages (early and late). Bacterial richness increased across succession in bulk soil and leaf endosphere. In contrast, soil fungal richness remained constant while root endosphere fungal richness increased across succession. There was, however, no significant difference in Shannon diversity indices between early and late successional stage in any compartment. There was a significant difference in the composition of microbial communities between early and late successional stage in all compartments, although the major microbial OTUs were shared between early and late successional stage. Co‐occurrence network analysis revealed successional stage‐specific microbial groups. There were more co‐occurring modules in early successional stage than in late stage. Altogether, these results emphasize that succession strongly affects distribution of microbial species, but not microbial diversity in arctic sand dune ecosystem and that fungi and bacteria may not follow the same successional trajectories.     

Occurrence of mites and fungi in the homes of patients with allergic manifestations

This survey was carried out to evaluate mites and moulds concentrations in the homes of patients with allergic manifestations and positive skin-test for mites and/or fungi. The home environments of 277 patients were evaluated by questionnaire, by sampling of airborne fungal spores and by determining the occurrence of mites and moulds in dust samples. Among the 277 patients examined, 83% reacted positively to house dust mites. The fungal allergen most frequently responsible for skin positivity was Alternaria tenuis. The prevalent airborne fungi were Cladosporium, Penicillium and Alternaria, followed by Aspergillus and Aureobasidium. The other genera were found in less than 50% of the homes. The presence of domestic mites in dust was documented in more than 85% of the homes sampled. The occurrence of fungi in dust was generally higher than in the air. The most common genera recovered were Penicillium, Candida, Aspergillus, Aureobasidium, Alternaria and Cladosporium. The homes investigated were divided into two groups, damp and dry, on the basis of the reported presence of wet or damp spots on inside surfaces and moulds growing inside the home. Our results showed that the concentration of mites and moulds in dust was significantly higher (p < 0.01) in damp homes as compared to dry ones.In general, a good agreement between skin positivity and presence of the same allergens in patients' home environment was observed for mites and, among fungi, for Alternaria, Cladosporium and Aspergillus. Our results seem to confirm that the study of mycoaeroflora and the quantification of mites and moulds in dust samples of allergic patients' homes could lead to more precise diagnoses and therefore to better prophylactic and therapeutic programs for each individual patient.     

Metagenomic assessment of the global diversity and distribution of bacteria and fungi

Bacteria and fungi are of uttermost importance in determining environmental and host functioning. Despite close interactions between animals, plants, their associated microbiomes, and the environment they inhabit, the distribution and role of bacteria and especially fungi across host and environments as well as the cross-habitat determinants of their community compositions remain little investigated. Using a uniquely broad global dataset of 13 483 metagenomes, we analysed the microbiome structure and function of 25 host-associated and environmental habitats, focusing on potential interactions between bacteria and fungi. We found that the metagenomic relative abundance ratio of bacteria-to-fungi is a distinctive microbial feature of habitats. Compared with fungi, the cross-habitat distribution pattern of bacteria was more strongly driven by habitat type. Fungal diversity was depleted in host-associated communities compared with those in the environment, particularly terrestrial habitats, whereas this diversity pattern was less pronounced for bacteria. The relative gene functional potential of bacteria or fungi reflected their diversity patterns and appeared to depend on a balance between substrate availability and biotic interactions. Alongside helping to identify hotspots and sources of microbial diversity, our study provides support for differences in assembly patterns and processes between bacterial and fungal communities across different habitats.     

Molecular analysis of environmental plant DNA in house dust across the United States

Despite the prevalence and costs of allergic diseases caused by pollen, we know little about the distributions of allergenic and non-allergenic pollen inside and outside homes at the continental scale. To better understand patterns in potential pollen diversity across the United States, we used DNA sequencing of a chloroplast marker gene to identify the plant DNA found in settled dust collected on indoor and outdoor surfaces across 459 homes. House location was the best predictor of the relative abundance of plant taxa found in outdoor dust samples. Urban, southern houses in hotter climates that were further from the coast were more likely to have more DNA from grass and moss species, while rural houses in northern, cooler climates closer to the coast were more likely to have higher relative abundances of DNA from Pinus and Cedrus species. In general, those plant taxa that were more abundant outdoors were also more abundant indoors, but indoor dust had uniquely high abundances of DNA from food plants and plants associated with lawns. Approximately 14 % of the plant DNA sequences found outside were from plant taxa that are known to have allergenic pollen compared to just 8 % inside. There was little geographic pattern in the total relative abundance of these allergens highlighting the difficulties associated with trying to predict allergen exposures based on geographic location alone. Together, this work demonstrates the utility of using environmental DNA sequencing to reconstruct the distributions of plant DNA inside and outside buildings, an approach that could prove useful for better understanding and predicting plant allergen exposures.     

Metagenomic Insights into the Bioaerosols in the Indoor and Outdoor Environments of Childcare Facilities

Airborne microorganisms have significant effects on human health, and children are more vulnerable to pathogens and allergens than adults. However, little is known about the microbial communities in the air of childcare facilities. Here, we analyzed the bacterial and fungal communities in 50 air samples collected from five daycare centers and five elementary schools located in Seoul, Korea using culture-independent high-throughput pyrosequencing. The microbial communities contained a wide variety of taxa not previously identified in child daycare centers and schools. Moreover, the dominant species differed from those reported in previous studies using culture-dependent methods. The well-known fungi detected in previous culture-based studies (Alternaria, Aspergillus, Penicillium, and Cladosporium) represented less than 12% of the total sequence reads. The composition of the fungal and bacterial communities in the indoor air differed greatly with regard to the source of the microorganisms. The bacterial community in the indoor air appeared to contain diverse bacteria associated with both humans and the outside environment. In contrast, the fungal community was largely derived from the surrounding outdoor environment and not from human activity. The profile of the microorganisms in bioaerosols identified in this study provides the fundamental knowledge needed to develop public health policies regarding the monitoring and management of indoor air quality.     

Airborne and Grain Dust Fungal Community Compositions Are Shaped Regionally by Plant Genotypes and Farming Practices

Chronic exposure to airborne fungi has been associated with different respiratory symptoms and pathologies in occupational populations, such as grain workers. However, the homogeneity in the fungal species composition of these bioaerosols on a large geographical scale and the different drivers that shape these fungal communities remain unclear. In this study, the diversity of fungi in grain dust and in the aerosols released during harvesting was determined across 96 sites at a geographical scale of 560 km² along an elevation gradient of 500 m by tag-encoded 454 pyrosequencing of the internal transcribed spacer (ITS) sequences. Associations between the structure of fungal communities in the grain dust and different abiotic (farming system, soil characteristics, and geographic and climatic parameters) and biotic (wheat cultivar and previous crop culture) factors were explored. These analyses revealed a strong relationship between the airborne and grain dust fungal communities and showed the presence of allergenic and mycotoxigenic species in most samples, which highlights the potential contribution of these fungal species to work-related respiratory symptoms of grain workers. The farming system was the major driver of the alpha and beta phylogenetic diversity values of fungal communities. In addition, elevation and soil CaCO₃ concentrations shaped the alpha diversity, whereas wheat cultivar, cropping history, and the number of freezing days per year shaped the taxonomic beta diversity of these communities.     

Increased levels of markers of microbial exposure in homes with indoor storage of organic household waste

As part of environmental management policies in Europe, separate collection of organic household waste and nonorganic household waste has become increasingly common. As waste is often stored indoors, this policy might increase microbial exposure in the home environment. In this study we evaluated the association between indoor storage of organic waste and levels of microbial agents in house dust. The levels of bacterial endotoxins, mold beta(1-->3)-glucans, and fungal extracellular polysaccharides (EPS) of Aspergillus and Penicillium species were determined in house dust extracts as markers of microbial exposure. House dust samples were collected in 99 homes in The Netherlands selected on the basis of whether separated organic waste was present in the house. In homes in which separated organic waste was stored indoors for 1 week or more the levels of endotoxin, EPS, and glucan were 3.2-, 7.6-, and 4. 6-fold higher, respectively (all P < 0.05), on both living room and kitchen floors than the levels in homes in which only nonorganic residual waste was stored indoors. Increased levels of endotoxin and EPS were observed, 2.6- and 2.1-fold (P < 0.1), respectively, when separated organic waste was stored indoors for 1 week or less, whereas storage of nonseparated waste indoors had no effect on microbial agent levels (P > 0.2). The presence of textile floor covering was another major determinant of microbial levels (P < 0.05). Our results indicate that increased microbial contaminant levels in homes are associated with indoor storage of separated organic waste. These increased levels might increase the risk of bioaerosol-related respiratory symptoms in susceptible people.     

Indoor and outdoor airborne fungal propagule concentrations in Mexico City

Thirty homes of asthmatic adults located in Mexico City were examined to determine the predominant culturable fungi and the changes in their airborne concentrations. Fungi were cultured and identified microscopically from air samples collected in naturally ventilated homes, during both wet (July–August) and cool dry (November–December) seasons, and from settled dust from the same homes. Airborne dust from indoor yielded 99–4950 cfu m⁻³, and settled dust 10²–10⁶ cfu g⁻¹ on DG18 agar. The indoor geometric mean concentration of airborne fungi during the cool dry season was 460 cfu m⁻³ while in the wet season it was 141 cfu m⁻³. Similarly, numbers of airborne fungal propagules out of doors decreased 60% between the dry and wet season. In general, the total fungal concentrations in indoor air were less than 10³ cfu m⁻³ and a large proportion of them was collected in Stage-2 of the Andersen sampler. Moreover, the ratio between indoor and outdoor concentrations was <3:1. Five of the 30 sampled homes yielded >500 cfu m⁻³ of one genus, with up to 1493Cladosporium cfu m⁻³ or 2549Penicillium cfu m⁻³. Also, these two genera were predominant in both airborne and settled dust, and their concentrations were greater indoors than out, indicating a possible indoor source of fungal propagules. The predominant species wereCladosporium herbarum, Penicillium aurantiogriseum andP. chrysogenum. These results suggest that exposure to large concentrations of fungi occurs indoors and is associated with both seasons of the year and with particular home characteristics.     

Dispersal in microbes: fungi in indoor air are dominated by outdoor air and show dispersal limitation at short distances

The indoor microbiome is a complex system that is thought to depend on dispersal from the outdoor biome and the occupants'' microbiome combined with selective pressures imposed by the occupants'' behaviors and the building itself. We set out to determine the pattern of fungal diversity and composition in indoor air on a local scale and to identify processes behind that pattern. We surveyed airborne fungal assemblages within 1-month time periods at two seasons, with high replication, indoors and outdoors, within and across standardized residences at a university housing facility. Fungal assemblages indoors were diverse and strongly determined by dispersal from outdoors, and no fungal taxa were found as indicators of indoor air. There was a seasonal effect on the fungi found in both indoor and outdoor air, and quantitatively more fungal biomass was detected outdoors than indoors. A strong signal of isolation by distance existed in both outdoor and indoor airborne fungal assemblages, despite the small geographic scale in which this study was undertaken (<500 m). Moreover, room and occupant behavior had no detectable effect on the fungi found in indoor air. These results show that at the local level, outdoor air fungi dominate the patterning of indoor air. More broadly, they provide additional support for the growing evidence that dispersal limitation, even on small geographic scales, is a key process in structuring the often-observed distance–decay biogeographic pattern in microbial communities.     

Seasonal Changes in Endotoxin Exposure and Its Relationship to Exhaled Nitric Oxide and Exhaled Breath Condensate pH Levels in Atopic and Healthy Children

Endotoxin, a component of the cell walls of gram-negative bacteria, is a contaminant in organic dusts (house dust) and aerosols. In humans, small amounts of endotoxin may cause a local inflammatory response. Exhaled nitric oxide (eNO) levels, an inflammation indicator, are associated with the pH values of exhaled breath condensate (EBC). This study evaluated seasonal changes on indoor endotoxin concentrations in homes and the relationships between endotoxin exposure and eNO/EBC pH levels for healthy children and children with allergy-related respiratory diseases. In total, 34 children with allergy-related respiratory diseases and 24 healthy children were enrolled. Indoor air quality measurements and dust sample analysis for endotoxin were conducted once each season inside 58 surveyed homes. The eNO, EBC pH levels, and pulmonary function of the children were also determined. The highest endotoxin concentrations were on kitchen floors of homes of children with allergy-related respiratory diseases and healthy children, and on bedroom floors of homes of asthmatic children and healthy children. Seasonal changes existed in endotoxin concentrations in dust samples from homes of children with allergic rhinitis, with or without asthma, and in EBC pH values among healthy children and those with allergy-related respiratory diseases. Strong relationships existed between endotoxin exposure and EBC pH values in children with allergic rhinitis.     

Do bacterial and fungal communities assemble differently during primary succession?

High‐throughput sequencing technologies are now allowing us to study patterns of community assembly for diverse microbial assemblages across environmental gradients and during succession. Here we discuss potential explanations for similarities and differences in bacterial and fungal community assembly patterns along a soil chronosequence in the foreland of a receding glacier. Although the data are not entirely conclusive, they do indicate that successional trajectories for bacteria and fungi may be quite different. Recent empirical and theoretical studies indicate that smaller microbes (like most bacteria) are less likely to be dispersal limited than are larger microbes – which could result in a more deterministic community assembly pattern for bacteria during primary succession. Many bacteria are also better adapted (than are fungi) to life in barren, early‐successional sediments in that some can fix nitrogen and carbon from the atmosphere – traits not possessed by any fungi. Other differences between bacteria and fungi are discussed, but it is apparent from this and other recent studies of microbial succession that we are a long way from understanding the mechanistic underpinnings of microbial community assembly during ecosystem succession. We especially need a better understanding of global and regional patterns of microbial dispersal and what environmental factors control the development of microbial communities in complex natural systems.     

Sapwood and heartwood affect differentially bacterial and fungal community structure and successional dynamics during Quercus petraea decomposition

In forests, bacteria and fungi are key players in wood degradation. Still, studies focusing on bacterial and fungal successions during the decomposition process depending on the wood types (i.e. sapwood and heartwood) remain scarce. This study aimed to understand the effect of wood type on the dynamics of microbial ecological guilds in wood decomposition. Using Illumina metabarcoding, bacterial and fungal communities were monitored every 3 months for 3 years from Quercus petraea wood discs placed on forest soil. Wood density and microbial enzymes involved in biopolymer degradation were measured. We observed rapid changes in the bacterial and fungal communities and microbial ecological guilds associated with wood decomposition throughout the experiment. Bacterial and fungal succession dynamics were very contrasted between sapwood and heartwood. The initial microbial communities were quickly replaced by new bacterial and fungal assemblages in the sapwood. Conversely, some initial functional guilds (i.e. endophytes and yeasts) persisted all along the experiment in heartwood and finally became dominant, possibly limiting the development of saprotrophic fungi. Our data also suggested a significant role of bacteria in nitrogen cycle during wood decomposition.     

Differentiations of determinants for the community compositions of bacteria,fungi, and nitrogen fixers in various steppes

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Airborne fungi as biocontaminants at two Milan underground stations

The viable or culturable air spora of two Milan underground stations and of the squares above has been investigated during a one-year study. Aerobiological data were collected by semi-quantitative method. Outdoor results show the presence of four dominant genera, Cladosporium, Penicillium, Epicoccum and Alternaria, whose presence varied throughout the year. Cryptococcus neoformans was isolated in Duomo Square. Indoor sampling shows to what extent the mycoflora of an indoor environment can depend both on the fungal spora coming from outside and the capacity of the fungi to colonize the different sublayers found indoors.