High indoor microbial levels are associated with reduced Th1 cytokine secretion capacity in infancy.

BACKGROUND: Exposure to microbes and their components may affect the maturation of the immune system. We examined the association of house dust microbial content with cytokine-producing capacity at birth and at the age of 1 year. METHODS: Production of TNF-α, IFN-γ, IL-5, IL-8 and IL-10 at birth (n = 228) and at the age of 1 year (n = 200) following 24- and 48-hour whole-blood stimulation with staphylococcal enterotoxin B (SEB), lipopolysaccharide and the combination of phorbol ester and ionomycin was measured. Concentrations of ergosterol (marker for fungal biomass), muramic acid (marker for Gram-positive bacteria) and 3-hydroxy fatty acids with a carbon chain length from 10 to 14 (marker for Gram-negative bacteria) in living room floor dust were analyzed using gas chromatography-tandem mass spectrometry. Five single microbial species or groups were determined using a quantitative polymerase chain reaction method. RESULTS: A high total level of the studied Gram-positive bacteria in general or Mycobacterium spp. in house dust was associated with decreased SEB-stimulated IFN-γ production, especially at the age of 1 year. The total level of indoor fungi analyzed (Penicillium spp., Aspergillus spp. and Paecilomyces variotii group, Trichoderma viride/atroviride/koningii,Wallemia sebi) was also inversely associated with IFN-γ production at the age of 1 year, but this association did not remain significant after adjustment for potential confounders. A few associations were found between microbial exposures and other measured cytokines. CONCLUSIONS: High indoor microbial exposures may affect immune development in early life by reducing T helper type 1 cytokine secretion capacity. The observed hyporesponsiveness may reflect the adaptation of the immune system to environmental antigens. In future, more attention should be paid especially to the immunomodulatory role of exposures to Gram-positive bacteria.

Microbial secondary metabolites in school buildings inspected for moisture damage in Finland, The Netherlands and Spain.

Secondary metabolites produced by fungi and bacteria are among the potential agents that contribute to adverse health effects observed in occupants of buildings affected by moisture damage, dampness and associated microbial growth. However, few attempts have been made to assess the occurrence of these compounds in relation to moisture damage and dampness in buildings. This study conducted in the context of the HITEA project (Health Effects of Indoor Pollutants: Integrating microbial, toxicological and epidemiological approaches) aimed at providing systematic information on the prevalence of microbial secondary metabolites in a large number of school buildings in three European countries, considering both buildings with and without moisture damage and/or dampness observations. In order to address the multitude and diversity of secondary metabolites a large number of more than 180 analytes was targeted in settled dust and surface swab samples using liquid chromatography/mass spectrometry (LC/MS) based methodology. While 42%, 58% and 44% of all samples collected in Spanish, Dutch and Finnish schools, respectively, were positive for at least one of the metabolites analyzed, frequency of detection for the individual microbial secondary metabolites - with the exceptions of emodin, certain enniatins and physcion - was low, typically in the range of and below 10% of positive samples. In total, 30 different fungal and bacterial secondary metabolites were found in the samples. Some differences in the metabolite profiles were observed between countries and between index and reference school buildings. A major finding in this study was that settled dust derived from moisture damaged, damp schools contained larger numbers of microbial secondary metabolites at higher levels compared to respective dust samples from schools not affected by moisture damage and dampness. This observation was true for schools in each of the three countries, but became statistically significant only when combining schools from all countries and thus increasing the sample number in the statistical analyses.

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.

Odour annoyance and physical symptoms among residents living near waste treatment centres.

Waste treatment processes produce odours and biological emissions to the environment, but their health effects are controversial. The aim of our study was to assess odour-associated self-reported physical symptoms among residents living near waste treatment centres. The study was conducted in the surroundings of five large-scale Finnish waste treatment centres with composting plants. In 2006, 1142 randomly selected residents living within 1.5, 3.0 and 5.0 km of these centres were interviewed by telephone. A questionnaire with 102 items asked about respondent's personal characteristics, odour exposure and symptoms during the preceding 12 months. Physical symptoms were analysed by distance to the waste treatment centre and by the respondent's perception and annoyance of waste treatment odour. The residents who were classified as "annoyed of the odour" reported following physical symptoms more than the others did: unusual shortness of breath (OR 1.5, 95% CI 1.0-2.2), eye irritation (1.5, 1.1-2.1), hoarseness/dry throat (1.5, 1.1-2.0), toothache (1.4, 1.0-2.1), unusual tiredness (1.5, 1.1-2.0), fever/shivering (1.7, 1.1-2.5), joint pain (1.5, 1.1-2.1) and muscular pain (1.5, 1.1-2.0). Moreover, the ORs for almost all other physical symptoms were elevated among the annoyed respondents. Reported odour annoyance near the waste treatment centres showed an association with many physical symptoms among residents living in the neighbouring areas.

Concentrations and diversity of microbes from four local bioaerosol emission sources in Finland.

Microbial particles can readily be released into the air from different types of man-made sources such as waste operations. Microbiological emissions from different biological sources and their dispersion may be an issue of concern for area planning and for nearby residents. This study was designed to determine the concentrations and diversity of microbiological emissions from four different man-made source environments: waste center with composting windrows, sewage treatment plant, farming environment, and cattle manure spreading. Samples of airborne particles were collected onto polyvinyl chloride filters at three distances along the prevailing downwind direction, from each source environment during a period of approximately 1 week. These samples were analyzed for 13 species or assay groups of fungi, bacterial genus Streptomyces, and Gram-positive and -negative bacteria using quantitative polymerase chain reaction (PCR). Samples for determining the concentrations of viable fungi and bacteria were collected from all environments using a six-stage impactor. The results show that there were variations in the microbial diversity between the source environments. Specifically, composting was a major source for the fungal genera Aspergillus and Penicillium, particularly for Aspergillus fumigatus, and for the bacterial genus Streptomyces. Although the microbial concentrations in the sewage treatment plant area were significantly higher than those at 50 or 200 m distance from the plant area, in the farming environment or cattle manure spreading area, no significant difference was observed between different distances from the source. In summary, elevated concentrations of microbes that differ from background can only be detected within a few hundred meters from the source. This finding, reported earlier for culturable bacteria and fungi, could thus be confirmed using molecular methods that cover both culturable and nonculturable microbial material.

Evaluating housing quality, health and safety using an Internet-based data collection and response system: a cross-sectional study.

BACKGROUND: Typically housing and health surveys are not integrated together and therefore are not representative of population health or national housing stocks. In addition, the existing channels for distributing information about housing and health issues to the general public are limited. The aim of this study was to develop a data collection and response system that would allow us to assess the Finnish housing stock from the points of view of quality, health and safety, and also to provide a tool to distribute information about important housing health and safety issues. METHODS: The data collection and response system was tested with a sample of 3000 adults (one per household), who were randomly selected from the Finnish Population Register Centre. Spatial information about the exact location of the residences (i.e. coordinates) was included in the database inquiry. People could participate either by completing and returning a paper questionnaire or by completing the same questionnaire via the Internet. The respondents did not receive any compensation for their time in completing the questionnaire. RESULTS: This article describes the data collection and response system and presents the main results of the population-based testing of the system. A total of 1312 people (response rate 44%) answered the questionnaire, though only 80 answered via the Internet. A third of the respondents had indicated they wanted feedback. Albeit a majority (>90%) of the respondents reported being satisfied or quite satisfied with their residence, there were a number of prevalent housing issues identified that can be related to health and safety. CONCLUSIONS: The collected database can be used to evaluate the quality of the housing stock in terms of occupant health and safety, and to model its association with occupant health and well-being. However, it must be noted that all the health outcomes gathered in this study are self-reported. A follow-up study is needed to evaluate whether the occupants acted on the feedback they received. Relying solely on an Internet-based questionnaire for collecting data would not appear to provide an adequate response rate for random population-based surveys at this point in time.

Odor annoyance near waste treatment centers: a population-based study in Finland.

Decomposition of biodegradable waste in municipal waste centers may produce odor emissions and subsequently cause discomfort to nearby residents. The public health importance of the resulting nuisance has not been sufficiently characterized. The aim of this study was to study the perception and annoyance of waste odor among residents in relation to distance from the large-scale source. In 2006, 1142 randomly selected residents living within 5 km from the boundaries of five waste treatment centers were interviewed by telephone. These centers were landfilling municipal waste and composting source-separated biowaste and/or sludge. The questionnaire consisted of 102 items containing questions on perceived environmental nuisance. Odds ratios (ORs) and confidence intervals (CIs) were calculated adjusting for sex and age. The proportion of respondents perceiving odor varied by center and distance (< 1.5 km: 66-100%; 1.5 to < 3 km: 13-84%; 3 to < 5 km: 2-64%). The pooled OR for odor annoyance was 6.1 (95% CI 3.7-10) in the intermediate and 19 (95% CI 12-32) in the innermost zone compared with residents in the outermost zone. Intensity of odor characterized as very strong or fairly strong affected odor annoyance more than weekly or more frequently perceived odor. The high level of odor perception and annoyance in residents living near waste treatment centers draws attention to the need to prevent odor nuisance constricting emission peaks and frequent emissions. Because odors may affect fairly distant (even 1.5 to < 3 km) residential areas, planning of the locations of waste treatment operations is essential.

Determination of bacterial load in house dust using qPCR, chemical markers and culture.

In this study, we developed two novel qPCR-assays for the detection of bacteria in house dust; one that determines the total bacterial amount and another that detects Gram-positive and Gram-negative bacteria separately. The methods were tested in silico and in vitro with microbial strains and vacuum cleaner dust samples, and validated in relation to culture and chemical marker analysis. We also compared the results of these three types of methods (qPCR, culture and chemical marker analysis) in 211 house dust samples from farming and non-farming environments. Microbial concentrations determined by the new qPCR assays (median 7.2 x 10(5) cell equivalents mg(-1)) were about two orders of magnitude higher than concentrations obtained by culture (median 6.7 x 10(3) cfu mg(-1)). The median concentration of muramic acid was 25.67 ng mg(-1) and that of 3-hydroxy fatty acids, expressed as LPS(10-16) was 26.14 pg mg(-1). Correlations between qPCR and chemical markers were moderate, while correlations between culture and qPCR and chemical markers were low to moderate. All the methods used in this study showed that the microbial concentrations are statistically significantly higher (p < 0.001, Mann-Whitney) in farming than non-farming environments.As a conclusion, all tested methods can be used for determining the bacterial load in dust samples, but none of the methods was superior to the others. The results obtained with these methods represent different aspects of bacterial exposure and therefore the results are not expected to be identical with each other.

Immunotoxicological properties of airborne particles at landfill, urban and rural sites and their relation to microbial concentrations.

We investigated the immunotoxicological activity of airborne particles in three different environments during 11 months. Specifically, we analyzed the relation of the immunotoxicological activity to microbial concentrations. During the study period, samples from a landfill, an urban and a rural site were collected on filters once a month. The immunotoxicological characteristics of collected particle samples were studied by exposing mouse macrophages (RAW264.7), and measuring the viability and production of inflammatory mediators i.e. nitric oxide (NO), tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 after 24 hours. In addition, the results were compared to corresponding microbial concentrations measured with quantitative polymerase chain reaction (qPCR) in the same sites. The particles collected from the landfill caused significantly more toxicity and inflammatory responses than the particles from other sites. The immunotoxicological activity of the samples changed according to the season, being the lowest in all study sites in the winter. In the rural and urban sites the responses peaked during the spring, whereas at the landfill the highest responses were detected towards the fall. All immunotoxic responses correlated strongly with airborne microbial concentrations at the landfill, whereas on the other sites the correlations were weaker. These results indicate that the overall immunotoxicological activity of the particles is increased in waste handling area with a heavy microbial load. The activity of the samples collected from different sites changes according to the season being at their lowest at wintertime. At the waste center, the immunotoxicological responses are related with concentrations of microbes, whereas at rural area and city center other factors seem to govern the toxicity of the sample.

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.

Predominance of Gram-positive bacteria in house dust in the low-allergy risk Russian Karelia.

Simple living conditions and farming environment have been associated with reduced risk for allergic diseases such as atopy and asthma but the factors responsible for this effect remain unresolved. We examined the bacterial composition of house dusts obtained from Finnish and Russian Karelia, two adjacent areas with high and low occurrence of atopic diseases respectively. Two dust mixes, both composed of 10 randomly selected dust samples from 349 Finnish and 417 Russian Karelian households were studied for bacterial biomarkers (DNA, Limulus-active endotoxin, 3-OH fatty acids, muramic acid) and for 16S rRNA gene sequences. Overall, the DNA cloning revealed more taxons (94 different genera) of dustborne bacteria than seen in any previous study on residential environments. Majority (67%) of the bacterial DNA clones in house dust from the low-allergy Russian Kareliarepresented Gram-positive bacteria (Firmicutes and Actinobacteria), predominantly Staphylococcaceae and Corynebacteriaceae. Russian Karelian dust showed up to 20-fold higher contents of muramic acid (marker of Gram-positive bacteria) and a sevenfold higher number of clones of animal-associated species, whereas in Finnish Karelian dust Gram-negatives (mainly Proteobacteria) predominated. Clones of plant-associated bacterial species and of chloroplast, indicating plant biomass, were more numerous in Finnish than in Russian Karelian dust. In conclusion, this study revealed major disparities between Finnish and Russian house dusts. The higher bacterial content and the predominance of Gram-positive bacteria in Russian dust may have implications for occurrence of atopy.

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.

Indoor air particles and bioaerosols before and after renovation of moisture-damaged buildings: the effect on biological activity and microbial flora.

Many building-related health problems coincide with moisture damage and mold growth within a building. Their elimination is assumed to improve indoor air quality. The aim of this study was to follow the success of remediation in two individual buildings by analyzing the microbial flora and immunotoxicological activity of filter samples. We compare results from samples collected from indoor air in the moisture-damaged buildings before and after renovation and results from matched reference buildings and outdoor air. The microbial characteristics of the samples were studied by analyzing ergosterol content and determining the composition of fungal flora with quantitative polymerase chain reaction (QPCR). In addition, the concentrations of particles were monitored with optical particle counter (OPC). The immunotoxicological activity of collected particle samples was tested by exposing mouse macrophages (RAW264.7) for 24 h to particle suspension extracted from the filters, and measuring the viability of the exposed cells (MTT-test) and production of inflammatory mediators (nitric oxide, IL-6 and TNF*) in cell culture medium by enzyme-linked immunoassay (ELISA). The results show that for Location 1 the renovation decreased the immunotoxicological activity of the particles collected from damaged building, whereas no difference was detected in the corresponding samples collected from the reference building. Interestingly, only slight differences were seen in the concentration of fungi. In the Location 2, a decrease was seen in the concentration of fungi after the renovation, whereas no effect on the immunotoxicological responses was detected. In this case, the immunotoxicological responses to the indoor air samples were almost identical to those caused by the samples from outdoor air. This indicates that the effects of remediation on the indoor air quality may not necessarily be readily measurable either with microbial or toxicological parameters. This may be associated with different spectrum of harmful agents in different mold and moisture-damaged buildings.

Monitoring success of remediation: seven case studies of moisture and mold damaged buildings.

Based on seven case studies of buildings that underwent different degrees of moisture and mold damage remediation, we aimed to develop methodology for assessment of the success of the remediation process. Methods used in gauging the success included technical monitoring of performance of building structures and heating, ventilation and air conditioning (HVAC) systems, microbial monitoring of indoor air quality (IAQ), and health effects studies of building occupants. The assessment was based on measurable change in the situations before and after remediation. Based on technical monitoring, remediation was successful in three cases, with partial improvement noted in three cases, whereas no remediation was conducted in one case. Based on microbial monitoring, improvement was detected in one, partial improvement in two and no improvement in two cases, whereas no follow-up was conducted in two cases. Health effect studies (mainly self-reported health status) showed improvement in one case, partial improvement in two cases, and no improvement in two cases, whereas no follow-up was conducted in one case, and in one case, follow-up failed due to low response rate. The results illustrate that it is possible to monitor the effects of remediation using various metrics. However, in some cases, no improvement could be observed in IAQ or occupant health, even if the remediation was considered technically successful, i.e. the remediation was fully completed as recommended. This could be due to many reasons, including: 1) all damage may not have been addressed adequately; 2) IAQ or health may not have been perceived improved regardless of remediation; and/or 3) the methods used may not have been sensitive/specific enough to detect such improvement within the 6-12 months follow-up periods after completion of the remediation. There is a need to further develop tools for monitoring and assessment of the success of moisture damage remediation in buildings.

Elements that contribute to healthy building design.

BACKGROUND: The elements that contribute to a healthy building are multifactorial and can be discussed from different perspectives. OBJECTIVES: WE PRESENT THREE VIEWPOINTS OF DESIGNING A HEALTHY BUILDING: the importance of sustainable development, the role of occupants for ensuring indoor air quality, and ongoing developments related to indoor finishes with low chemical emissions and good fungal resistance. DISCUSSION: Sustainable design rediscovers the social, environmental, and technical values of pedestrian and mixed-use communities, using existing infrastructures including "main streets" and small-town planning principles and recapturing indoor-outdoor relationships. This type of design introduces nonpolluting materials and assemblies with lower energy requirements and higher durability and recyclability. Building occupants play a major role in maintaining healthy indoor environments, especially in residences. Contributors to indoor air quality include cleaning habits and other behaviors; consumer products, furnishings, and appliances purchases, as well as where and how the occupants use them. Certification of consumer products and building materials as low-emitting products is a primary control measure for achieving good indoor air quality. Key products in this respect are office furniture, flooring, paints and coatings, adhesives and sealants, wall coverings, wood products, textiles, insulation, and cleaning products. Finishing materials play a major role in the quality of indoor air as related to moisture retention and mold growth. CONCLUSIONS: Sustainable design emphasizes the needs of infrastructure, lower energy consumption, durability, and recyclability. To ensure good indoor air quality, the product development for household use should aim to reduce material susceptibility to contaminants such as mold and should adopt consumer-oriented product labeling.

Comparison of mold concentrations quantified by MSQPCR in indoor and outdoor air sampled simultaneously.

Mold specific quantitative PCR (MSQPCR) was used to measure the concentrations of the 36 mold species in indoor and outdoor air samples that were taken simultaneously for 48 h in and around 17 homes in Cincinnati, Ohio. The total spore concentrations of 353 per m(3) of indoor air and 827 per m(3) of outdoor air samples were significantly different (por=0.5). These results suggest that interpretation of the meaning of short-term (<48 h) mold measurements in indoor and outdoor air samples must be made with caution.

Microbial exposure, symptoms and inflammatory mediators in nasal lavage fluid of kitchen and clerical personnel in schools.

OBJECTIVES: The aim of this study was to investigate how the microbial conditions of kitchen facilities differ from those in other school facilities. The health status of the personnel was also studied. MATERIALS AND METHODS: The microbial investigations were conducted in six moisture-damaged schools and two reference schools. The symptoms of the kitchen personnel were surveyed with questionnaires and inflammatory responses in nasal lavage (NAL) fluid were measured. RESULTS: The total concentrations of airborne microbes were lower in kitchens than in other facilities of the schools. However, the occurrence of moisture damage increased the airborne microbial concentrations both in kitchens, and in other facilities. Bacterial concentrations were high on surfaces in the damaged kitchens. Gram-negative bacteria predominated, but also thermophilic bacteria and mycobacteria were detected. Respiratory and general symptoms were prevalent both among kitchen workers and clerical personnel in the moisture-damaged environments. Reported allergies and repeated respiratory infections were connected with high IL-4 concentrations in NAL fluid. Median concentrations of studied inflammatory mediators (NO, IL-4, IL-6 and TNF-alpha) were slightly higher in NAL samples of kitchen workers than among the clerical personnel. CONCLUSIONS: Kitchen facilites differ from other facilities of the school building for their moisture conditions and microbial contamination. Thus, they represent a specific type of environment that may affect the health status of the personnel.

Production of proinflammatory mediators by indoor air bacteria and fungal spores in mouse and human cell lines.

We compared the inflammatory and cytotoxic responses caused by household mold and bacteria in human and mouse cell lines. We studied the fungi Aspergillus versicolor, Penicillium spinulosum, and Stachybotrys chartarum and the bacteria Bacillus cereus, Pseudomonas fluorescens, and Streptomyces californicus for their cytotoxicity and ability to stimulate the production of inflammatory mediators in mouse RAW264.7 and human 28SC macrophage cell lines and in the human A549 lung epithelial cell line in 24-hr exposure to 10(5), 10(6), and 10(7) microbes/mL. We studied time dependency by terminating the exposure to 10(6) microbes/mL after 3, 6, 12, 24, and 48 hr. We analyzed production of the cytokines tumor necrosis factor-alpha and interleukins 6 and 1ss (TNF-alpha, IL-6, IL-1ss, respectively) and measured nitric oxide production using the Griess method, expression of inducible NO-synthase with Western Blot analysis, and cytotoxicity with the MTT-test. All bacteria strongly induced the production of TNF-alpha, IL-6 and, to a lesser extent, the formation of IL-1ss in mouse macrophages. Only the spores of Str. californicus induced the production of NO and IL-6 in both human and mouse cells. In contrast, exposure to fungal strains did not markedly increase the production of NO or any cytokine in the studied cell lines except for Sta. chartarum, which increased IL-6 production somewhat in human lung epithelial cells. These microbes were less cytotoxic to human cells than to mouse cells. On the basis of equivalent numbers of bacteria and spores of fungi added to cell cultures, the overall potency to stimulate the production of proinflammatory mediators decreased in the order Ps. fluorescens > Str. californicus > B. cereus > Sta. chartarum > A. versicolor > P. spinulosum. These data suggest that bacteria in water-damaged buildings should also be considered as causative agents of adverse inflammatory effects.

Media for cultivation of indoor streptomycetes.

The growth of 10 indoor Streptomyces spp. isolates on nutritionally complex and selective 26 media revealed that the mycelium production had a tendency to increase in order: starch-casein < glycerol-arginine < glucose-tryptone, and NH(4)NO(3) < Na-caseinate-asparagine. Yeast extract increased mycelium biosynthesis, but not always the growth rate. The strains belonging to streptomycetes most common environmental isolates produced visible mycelium in 5 days on all media.