Microbial communities associated with house dust.

House dust is a complex mixture of inorganic and organic material with microbes in abundance. Few microbial species are actually able to grow and proliferate in dust and only if enough moisture is provided. Hence, most of the microbial content originates from sources other than the dust itself. The most important sources of microbes in house dust are outdoor air and other outdoor material tracked into the buildings, occupants of the buildings including pets and microbial growth on moist construction materials. Based on numerous cultivation studies, Penicillium, Aspergillus, Cladosporium, and about 20 other fungal genera are the most commonly isolated genera from house dust. The cultivable bacterial flora is dominated by Gram-positive genera, such as Staplylococcus, Corynebacterium, and Lactococcus. Culture-independent studies have shown that both the fungal and the bacterial flora are far more diverse, with estimates of up to 500-1000 different species being present in house dust. Concentrations of microbes in house dust vary from nondetectable to 10(9) cells g(-1) dust, depending on the dust type, detection method, type of the indoor environment and season, among other factors. Microbial assemblages in different house dust types usually share the same core species; however, alterations in the composition are caused by differing sources of microbes for different dust types. For example, mattress dust is dominated by species originating from the user of the mattress, whereas floor dust reflects rather outdoor sources. Farming homes contain higher microbial load than urban homes and according to a recent study, temperate climate zones show higher dust microbial diversity than tropical zones.

Molecular profiling of fungal communities in moisture damaged buildings before and after remediation--a comparison of culture-dependent and culture-independent methods.

BACKGROUND: Indoor microbial contamination due to excess moisture is an important contributor to human illness in both residential and occupational settings. However, the census of microorganisms in the indoor environment is limited by the use of selective, culture-based detection techniques. By using clone library sequencing of full-length internal transcribed spacer region combined with quantitative polymerase chain reaction (qPCR) for 69 fungal species or assay groups and cultivation, we have been able to generate a more comprehensive description of the total indoor mycoflora. Using this suite of methods, we assessed the impact of moisture damage on the fungal community composition of settled dust and building material samples (n = 8 and 16, correspondingly). Water-damaged buildings (n = 2) were examined pre- and post- remediation, and compared with undamaged reference buildings (n = 2). RESULTS: Culture-dependent and independent methods were consistent in the dominant fungal taxa in dust, but sequencing revealed a five to ten times higher diversity at the genus level than culture or qPCR. Previously unknown, verified fungal phylotypes were detected in dust, accounting for 12% of all diversity. Fungal diversity, especially within classes Dothideomycetes and Agaricomycetes tended to be higher in the water damaged buildings. Fungal phylotypes detected in building materials were present in dust samples, but their proportion of total fungi was similar for damaged and reference buildings. The quantitative correlation between clone library phylotype frequencies and qPCR counts was moderate (r = 0.59, p < 0.01). CONCLUSIONS: We examined a small number of target buildings and found indications of elevated fungal diversity associated with water damage. Some of the fungi in dust were attributable to building growth, but more information on the material-associated communities is needed in order to understand the dynamics of microbial communities between building structures and dust. The sequencing-based method proved indispensable for describing the true fungal diversity in indoor environments. However, making conclusions concerning the effect of building conditions on building mycobiota using this methodology was complicated by the wide natural diversity in the dust samples, the incomplete knowledge of material-associated fungi fungi and the semiquantitative nature of sequencing based methods.

The occupant as a source of house dust bacteria.

BACKGROUND: Markers for microbial groups are commonly measured in house dust samples to assess indoor exposure to microbes in studies on asthma and allergy. However, little is known about the sources of different microbes. A better understanding of the nature and origin of microbes present in the immediate environment of human beings is crucial if one wants to elucidate protective as well as adverse effects on human health. OBJECTIVE: To determine the extent to which the bacterial composition of mattress and floor dust reflects the presence of the human body in relation to other environmental sources. METHODS: House dust and skin surface swab samples of occupants in 4 homes were collected and analyzed for their bacterial content, using a culture-independent methodology. Bacterial sequences analyzed from the different house dusts and skin surface swabs represented random samples of bacteria present in a given sample. Highly similar sequences were grouped to assess biodiversity and to draw conclusions about the sources of bacteria. RESULTS: The bacterial flora in the house dust samples was found to be highly diverse and dominated by gram-positive bacteria. To a considerable extent, the presence of different bacterial groups was attributed to human sources. In the individuals' mattress dust samples, 69% to 88% of the bacterial sequences analyzed were associated with human origins. The respective percentages for the individual floor dusts ranged from 45% to 55%. CONCLUSION: Our study indicates that human-derived bacteria account for a large part of the mainly gram-positive bacterial content in house dust.

Probe-based negative selection for underrepresented phylotypes in large environmental clone libraries.

Studies based on cloning and sequencing to investigate microbial diversity in a vast range of samples has become widespread in recent years. Results have revealed immense microbial diversity in many different environments, but also dominance of a few sequence types in the constructed clone libraries. Here we describe a method to enrich the clone libraries by avoiding sequencing of known, abundant sequence types, instead focusing on novel, rare ones. The protocol is based on gridding the PCR products from clone libraries on membranes and hybridisation of species-specific probes. Clones that do not give positive hybridisation results are sequenced. This method was used for fungal clone libraries from compost samples. Altogether 1536 clones were gridded and six probes used. From these clones, 59% hybridised with a probe, and therefore, only 41% of the clones were sequenced. In addition, 384 samples were sequenced to verify the hybridisation results. The numbers of false-negative (5.2%) and false-positive (3.9%) hybridisations were low. This method provides a mean of lowering the costs of sequencing projects and speeding up the process of characterising microbial diversity in environmental samples. The method is especially suitable for samples with a few dominating sequence types.

Diversity and seasonal dynamics of bacterial community in indoor environment.

BACKGROUND: We spend most of our lives in indoor environments and are exposed to microbes present in these environments. Hence, knowledge about this exposure is important for understanding how it impacts on human health. However, the bacterial flora in indoor environments has been only fragmentarily explored and mostly using culture methods. The application of molecular methods previously utilised in other environments has resulted in a substantial increase in our awareness of microbial diversity. RESULTS: The composition and dynamics of indoor dust bacterial flora were investigated in two buildings over a period of one year. Four samples were taken in each building, corresponding to the four seasons, and 16S rDNA libraries were constructed. A total of 893 clones were analysed and 283 distinct operational taxonomic units (OTUs) detected among them using 97% sequence similarity as the criterion. All libraries were dominated by Gram-positive sequences, with the most abundant phylum being Firmicutes. Four OTUs having high similarity to Corynebacterium-, Propionibacterium-, Streptococcus- and Staphylococcus- sequences were present in all samples. The most abundant of the Gram-negative OTUs were members of the family Sphingomonadaceae, followed by Oxalobacteraceae, Comamonadaceae, Neisseriaceae and Rhizobiaceae. The relative abundance of alpha- and betaproteobacteria increased slightly towards summer at the expense of firmicutes. The proportion of firmicutes and gammaproteobacteria of the total diversity was highest in winter and that of actinobacteria, alpha- and betaproteobacteria in spring or summer, whereas the diversity of bacteroidetes peaked in fall. A statistical comparison of the libraries revealed that the bacterial flora of the two buildings differed during all seasons except spring, but differences between seasons within one building were not that clear, indicating that differences between the buildings were greater than the differences between seasons. CONCLUSION: This work demonstrated that the bacterial flora of indoor dust is complex and dominated by Gram-positive species. The dominant phylotypes most probably originated from users of the building. Seasonal variation was observed as proportional changes of the phyla and at the species level. The microflora of the two buildings investigated differed statistically and differences between the buildings were more pronounced than differences between seasons.