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Antibodies to molds and satratoxin in individuals exposed in water-damaged buildings.

Immunoglobulin (Ig)A, IgM, and IgG antibodies against Penicillium notatum, Aspergillus niger, Stachybotrys chartarum, and satratoxin H were determined in the blood of 500 healthy blood donor controls, 500 random patients, and 500 patients with known exposure to molds. The patients were referred to the immunological testing laboratory for health reasons other than mold exposure, or for measurement of mold antibody levels. Levels of IgA, IgM, and IgG antibodies against molds were significantly greater in the patients (p < 0.001 for all measurements) than in the controls. However, in mold-exposed patients, levels of these antibodies against satratoxin differed significantly for IgG only (p < 0.001), but not for IgM or IgA. These differences in the levels of mold antibodies among the 3 groups were confirmed by calculation of z score and by Scheffé's significant difference tests. A general linear model was applied in the majority of cases, and 3 different subsets were formed, meaning that the healthy control groups were different from the random patients and from the mold-exposed patients. These findings indicated that mold exposure was more common in patients who were referred for immunological evaluation than it was in healthy blood donors. The detection of antibodies to molds and satratoxin H likely resulted from antigenic stimulation of the immune system and the reaction of serum with specially prepared mold antigens. These antigens, which had high protein content, were developed in this laboratory and used in the enzyme-linked immunosorbent assay (ELISA) procedure. The authors concluded that the antibodies studied are specific to mold antigens and mycotoxins, and therefore could be useful in epidemiological and other studies of humans exposed to molds and mycotoxins.

Adult↗

Health symptoms caused by molds in a courthouse.

A majority of occupants of a newly renovated historic courthouse in Calgary, Alberta, Canada, reported multiple (3 or more) health-related symptoms, and several reported more than 10 persistent symptoms. Most required at least 1 day outside of the building to recover from their symptoms. Molds that produce mycotoxins, such as Stachybotrys chartarum and Emericella nidulans, were identified in the building, along with fungal organisms of the genera Aspergillus, Penicillium, Streptomyces, Cladosporium, Chaetomium, Rhizopus/Mucor, Alternaria, Ulocladium, and Basidiomycetes. Renovations to this historic had building failed to provide adequate thermal and vapor barriers, thus allowing moist indoor air to migrate into the building enclosure, causing condensation to develop. Mold grew on the condensation and was dispersed throughout the courthouse, including on furniture and files. The courthouse was closed and a new facility was modified with low-offgassing materials, better ventilation and air filtration, and strict building maintenance to accommodate those occupants of the older building who had developed multiple chemical sensitivities.

Air Pollutants↗

Evaluation of bioaerosol exposures during conditioning of biofilter organic media beds.

Biological media air filters (biofilters) are currently being used for the treatment of inorganic and organic gases from sewage treatment plants, industrial processes, and remediation systems. The media may be organic material such as compost, wood chips, or synthetic plastic media, each with a large surface area for microorganism growth and activity. An occupational health and safety graduate student team (OHS team) evaluated potential particulate and bioaerosol exposure from a biofilter unit process used to treat hydrogen sulfide (H2S) gas generated from a primary sludge settling unit process. The OHS team included an industrial hygiene/environmental health engineering specialist, an occupational safety specialist, an occupational health physician, and an occupational health nurse. Concerns were raised regarding the possibility of adverse health effects to maintenance workers during "conditioning" of the biofilter compost-like media beds. Conditioning activities may include in-situ rearrangement of the existing media, removal from the tank/surface, drying/reinsertion of the existing media, or complete removal of the media, and replacement with new. Neither the design engineering firm nor the manufacturer had specific written recommendations or precautions regarding exposure during the conditioning of the compost beds. No personal protection equipment has been used for this activity. The expected agents for adverse health effects associated with this unit process are respirable particulate dust and bioaerosols, which may contain viable bacteria and fungi, as well as endotoxin. Safety procedures are already in place for H2S. Mixed dust from the compost media bed may cause irritation of pre-existing health conditions such as asthma, chronic lung disease, and some skin conditions, and may also lead to new health problems such as inhalation fever, occupational asthma, hypersensitivity pneumonitis, skin rashes and/or skin infections, and upper or lower respiratory infections. Air samples were taken immediately before (background samples) and during a simulation of the conditioning procedure of one of the two compost beds. Airborne samples collected during the simulated procedure yielded more bacteria per volume of collected air than the background samples. These findings suggest an increased risk for bacterial infection during the conditioning procedure. The airborne samples cultured for fungi during the simulation were less than the background sample. Stachybotrys chartarum was not detected in a sample of the filter media. As with the fungi data, endotoxin air sampling yielded less endotoxin during the simulated conditioning procedure than in the background. Two inches of rain had fallen in the 24 hours preceding the sampling, soaking the upper layers of the compost media. Sampling during an actual conditioning procedure on a warm, dry day may yield additional evidence relevant to potential health risks from dust and microorganisms. Although the data is not representative of an actual conditioning procedure, the following common sense precautions are recommended: (1) Dust minimization during dry conditions can be achieved by thoroughly wetting the compost bed prior to removing the compost, and wetting the compost again before returning it to the bed. (2) Based on scientific literature rather than this project's sampling results, the use of an appropriate respirator should be proactively considered. In addition, consideration must be given to the worker's cardiopulmonary effects associated with respirator use, decreased work performance effects, and OSHA administrative requirements of a respirator protection program. (3) Workers with immunocompromised status, asthma, allergic rhinitis/conjunctivitis, or allergic conditions of the skin, eyes, or lungs should be excluded from this particular job, unless it can be shown that their personal protective equipment is effective. (4) Protective outer wear, goggles, and gloves should be worn to minimize health risks associated with clothing garments contaminated by microorganisms.

Aerosols↗

Mold infestation of wet spray-applied cellulose insulation.

Mold investigations were conducted in four buildings that had been insulated with wet spray-applied cellulose insulation (WSACI). Bulk WSACI samples were collected and analyzed by quantitative polymerase chain reaction (QPCR) methods. Airborne mold was evaluated using both Burkard total mold spore and Andersen culturable/viable sampling methods. Although reportedly treated with biocidal borates, QPCR analyses indicated that elevated concentrations of mold cells (reported as spore equivalents per gram) may be present in WSACI. QPCR analyses showed the following: (1) very high concentrations of Penicillium chrysogenum in samples from two of four buildings; (2) very high concentrations of Stachybotrys chartarum in samples from one building and a more moderate presence in a second; (3) moderately high concentrations of Aspergillus versicolor in samples from one building and more moderate concentrations in a second; (4) the presence of the opportunistic pathogen, Aspergillus fumigatus, in samples from three of the four buildings, and (5) the presence of 22 of 23 target mold species. Elevated airborne total mold spore concentrations were observed in all four of the buildings investigated. Culturable/viable airborne mold concentrations were moderately elevated in three of the four buildings. Mold genera/types present were relatively consistent among airborne mold samples collected by both methods and bulk sample analyses. Results of this study suggest that WSACI has the potential to cause elevated airborne mold levels in buildings where it has been applied and pose significant mold exposure and public health risks.

Air Microbiology↗

Airborne fungal fragments and allergenicity.

Exposure to fungi, particularly in water damaged indoor environments, has been thought to exacerbate a number of adverse health effects, ranging from subjective symptoms such as fatigue, cognitive difficulties or memory loss to more definable diseases such as allergy, asthma and hypersensitivity pneumonitis. Understanding the role of fungal exposure in these environments has been limited by methodological difficulties in enumerating and identifying various fungal components in environmental samples. Consequently, data on personal exposure and sensitization to fungal allergens are mainly based on the assessment of a few select and easily identifiable species. The contribution of other airborne spores, hyphae and fungal fragments to exposure and allergic sensitization are poorly characterized. There is increased interest in the role of aerosolized fungal fragments following reports that the combination of hyphal fragments and spore counts improved the association with asthma severity. These fragments are particles derived from any intracellular or extracellular fungal structure and are categorized as either submicron particles or larger fungal fragments. In vitro studies have shown that submicron particles of several fungal species are aerosolized in much higher concentrations (300-500 times) than spores, and that respiratory deposition models suggest that such fragments of Stachybotrys chartarum may be deposited in 230-250 fold higher numbers than spores. The practical implications of these models are yet to be clarified for human exposure assessments and clinical disease. We have developed innovative immunodetection techniques to determine the extent to which larger fungal fragments, including hyphae and fractured conidia, function as aeroallergen sources. These techniques were based on the Halogen Immunoassay (HIA), an immunostaining technique that detects antigens associated with individual airborne particles >1 microm, with human serum immunoglobulin E (IgE). Our studies demonstrated that the numbers of total airborne hyphae were often significantly higher in concentration than conidia of individual allergenic genera. Approximately 25% of all hyphal fragments expressed detectable allergen and the resultant localization of IgE immunostaining was heterogeneous among the hyphae. Furthermore, conidia of ten genera that were previously uncharacterized could be identified as sources of allergens. These findings highlight the contribution of larger fungal fragments as aeroallergen sources and present a new paradigm of fungal exposure. Direct evidence of the associations between fungal fragments and building-related disease is lacking and in order to gain a better understanding, it will be necessary to develop diagnostic reagents and detection methods, particularly for submicron particles. Assays using monoclonal antibodies enable the measurement of individual antigens but interpretation can be confounded by cross-reactivity between fungal species. The recent development of species-specific monoclonal antibodies, used in combination with a fluorescent-confocal HIA technique should, for the first time, enable the speciation of morphologically indiscernible fungal fragments. The application of this novel method will help to characterize the contribution of fungal fragments to adverse health effects due to fungi and provide patient-specific exposure and sensitization profiles.

Air Microbiology↗

Effect of building frame and moisture damage on microbiological indoor air quality in school buildings.

The effect of building frame and moisture damage on microbial indoor air quality was characterized in 17 wooden and 15 concrete or brick school buildings. Technical investigations to detect visible moisture and mold damage were performed according to a standardized protocol. Viable airborne microbes were determined by using a six-stage impactor (Andersen 10-800). Mean concentrations of viable airborne fungi were significantly higher in wooden schools than in concrete schools, showing that the frame material was a determinant of concentrations of airborne fungi. Moisture damage of the building did not alter the fungal concentrations in wooden school buildings. In contrast, in concrete schools the effect of moisture damage was clearly seen as higher concentrations compared with the reference schools. Aspergillus versicolor, Stachybotrys, and Acremonium were detected only in samples from moisture damaged buildings, and can be considered marker fungi of such damage in school buildings. In addition, the presence of Oidiodendron as well as elevated concentrations of Cladosporium and actinobacteria were associated with moisture damage in concrete schools.

Air Pollution, Indoor↗

An investigation into techniques for cleaning mold-contaminated home contents.

This study examined the efficacy of the following treatments to reduce selected fungal spore and mycotoxin levels on materials commonly found in home contents: (1) gamma irradiation at a 10-13 kiloGray exposure, (2) a detergent/bleach wash, and (3) a steam cleaning technique. A minimum of six replicates were performed per treatment. Paper, cloth, wood, and carpet were inoculated with either fungal spores (Stachybotrys chartarum, Aspergillus niger, Penicillium chrysogenum, or Chaetomium globosum) at 240,000 spores/2.54 cm2 of material or with the mycotoxins roridin A, T-2, and verrucarin A at 10 microg per 2.54 cm2 of material. Treatments were evaluated with an agar plating technique for fungal spores and a yeast toxicity culture assay for mycotoxins. Results showed that gamma irradiation inactivated fungal spores, but the treatment was not successful in inactivating mycotoxins. The washing technique completely inactivated or removed spores on all materials except for C. globosum, which was reduced on all items except paper (p < 0.05). Washing inactivated all mycotoxins on paper and cloth but not on carpet or untreated wood (p < 0.001). The steam cleaning treatment did not completely eliminate any fungal spores; however, it reduced P. chrysogenum numbers on all materials, C. globosum was reduced on wood and carpet, and S. chartarum was reduced on wood (p < 0.05). Steam cleaning was unsuccessful in inactivating any of the tested mycotoxins. These results show that the bleach/detergent washing technique was more effective overall in reducing spore and mycotoxin levels than gamma irradiation or steam cleaning. However, the other examined techniques were successful in varying degrees.

Chlorine↗

Controlled study of mold growth and cleaning procedure on treated and untreated wet gypsum wallboard in an indoor environment.

The basis for some common gypsum wallboard mold remediation practices was examined. The bottom inch of several gypsum wallboard panels was immersed in bottled drinking water; some panels were coated and others were untreated. The panels were examined and tested for a period of 8 weeks. This study investigated: (a) whether mold growth, detectable visually or with tape lift samples, occurs within 1 week on wet gypsum wallboard; (b) the types, timing, and extent of mold growth on wet gypsum wallboard; (c) whether mold growth is present on gypsum wallboard surfaces 6 inches from visible mold growth; (d) whether some commonly used surface treatments affect the timing of occurrence and rate of mold growth; and (e) if moldy but dried gypsum wallboard can be cleaned with simple methods and then sealed with common surface treatments so that residual mold particles are undetectable with typical surface sampling techniques. Mold growth was not detected visually or with tape lift samples after 1 week on any of the wallboard panels, regardless of treatment, well beyond the 24-48 hours often mentioned as the incubation period. Growth was detected at 2 weeks on untreated gypsum. Penicillium, Cladosporium, and Acremonium were early colonizers of untreated panels. Aspergillus, Epicoccum, Alternaria, and Ulocladium appeared later. Stachybotrys was not found. Mold growth was not detected more than 6 inches beyond the margin of visible mold growth, suggesting that recommendations to remove gypsum wallboard more than 1 foot beyond visible mold are excessive. The surface treatments resulted in delayed mold growth and reduced the area of mold growth compared with untreated gypsum wallboard. Results showed that simple cleaning of moldy gypsum wallboard was possible to the extent that mold particles beyond "normal trapping" were not found on tape lift samples. Thus, cleaning is an option in some situations where removal is not feasible or desirable. In cases where conditions are not similar to those of this study, or where large areas may be affected, a sample area could be cleaned and tested to verify that the cleaning technique is sufficient to reduce levels to background or normal trapping. These results are generally in agreement with laboratory studies of mold growth on, and cleaning of, gypsum wallboard.

Air Pollutants↗

An environmental assessment of mold concentrations and potential mycotoxin exposures in the greater Southeast Texas area.

An assessment of indoor air quality in homes from the greater Houston and Southeast Texas area was sampled over a three-month period to ascertain the potential toxic exposure to molds and mycotoxins. In 2001 Houston experienced widespread flooding due to Tropical Storm Allison. The slow receding waters allowed molds to proliferate to great levels and affected many homes and businesses in and around the Houston area. Source (air) and surface (swab, tape, and bulk) samples were taken in fungal contaminated residences to measure the amounts of airborne and surface contaminants. Indoor airborne counts of fungal spores that were identified to be 80% or more of the outside counts of similar fungal genera were classified as indoor air contaminants. This study identified species of Aspergillus, Penicilliulm, and Cladosporium at levels ranging from 80% to 20 times greater than the corresponding outdoor readings in the greater Southeast Texas area. Aspergillus and Penicillium species are considered potentially toxic to humans at elevated levels due to their ability to produce mycotoxins. Cladosporium is considered nonpathogenic, but can still cause allergenic reactions from hyphal fragments. Surface sampling which is utilized for identification of visible mold contaminants, identified species of Aspergillus. Penicillium, Chaetomium, Cladosporium, and Stachybotrys ranging from 1 to 100 total colony counts (1-100% for tape and bulk noncultured assays) contingent upon the extent of visual contamination. The numerical values from surface sampling were used primarily to identify the degree of visual contamination, as well as, identify any molds that may have been too heavy to become aerosolized.

Air Pollution, Indoor↗

Adverse human health effects associated with molds in the indoor environment.

Molds are common and important allergens. About 5% of individuals are predicted to have some allergic airway symptoms from molds over their lifetime. However, it should be remembered that molds are not dominant allergens and that the outdoor molds, rather than indoor ones, are the most important. For almost all allergic individuals, the reactions will be limited to rhinitis or asthma; sinusitis may occur secondarily due to obstruction. Rarely do sensitized individuals develop uncommon conditions such as ABPA or AFS. To reduce the risk of developing or exacerbating allergies, mold should not be allowed to grow unchecked indoors. When mold colonization is discovered in the home, school, or office, it should be remediated after the source of the moisture that supports its growth is identified and eliminated. Authoritative guidelines for mold remediation are available. Fungi are rarely significant pathogens for humans. Superficial fungal infections of the skin and nails are relatively common in normal individuals, but those infections are readily treated and generally resolve without complication. Fungal infections of deeper tissues are rare and in general are limited to persons with severely impaired immune systems. The leading pathogenic fungi for persons with nonimpaired immune function, Blastomyces, Coccidioides, Cryptococcus, and Histoplasma, may find their way indoors with outdoor air but normally do not grow or propagate indoors. Due to the ubiquity of fungi in the environment, it is not possible to prevent immunecompromised individuals from being exposed to molds and fungi outside the confines of hospital isolation units. Some molds that propagate indoors may under some conditions produce mycotoxins that can adversely affect living cells and organisms by a variety of mechanisms. Adverse effects of molds and mycotoxins have been recognized for centuries following ingestion of contaminated foods. Occupational diseases are also recognized in association with inhalation exposure to fungi, bacteria, and other organic matter, usually in industrial or agricultural settings. Molds growing indoors are believed by some to cause building-related symptoms. Despite a voluminous literature on the subject, the causal association remains weak and unproven, particularly with respect to causation by mycotoxins. One mold in particular, Stachybotrys chartarum, is blamed for a diverse array of maladies when it is found indoors. Despite its well-known ability to produce mycotoxins under appropriate growth conditions, years of intensive study have failed to establish exposure to S. chartarum in home, school, or office environments as a cause of adverse human health effects. Levels of exposure in the indoor environment, dose-response data in animals, and dose-rate considerations suggest that delivery by the inhalation route of a toxic dose of mycotoxins in the indoor environment is highly unlikely at best, even for the hypothetically most vulnerable subpopulations. Mold spores are present in all indoor environments and cannot be eliminated from them. Normal building materials and furnishings provide ample nutrition for many species of molds, but they can grow and amplify indoors only when there is an adequate supply of moisture. Where mold grows indoors there is an inappropriate source of water that must be corrected before remediation of the mold colonization can succeed. Mold growth in the home, school, or office environment should not be tolerated because mold physically destroys the building materials on which it grows, mold growth is unsightly and may produce offensive odors, and mold is likely to sensitize and produce allergic responses in allergic individuals. Except for persons with severely impaired immune systems, indoor mold is not a source of fungal infections. Current scientific evidence does not support the proposition that human health has been adversely affected by inhaled mycotoxins in home, school, or office environments.

Air Pollution, Indoor↗

Specific molds associated with asthma in water-damaged homes.

OBJECTIVE: We sought to determine if specific molds were found in significantly higher concentrations in the water-damaged homes of asthmatic children compared with homes with no visible water damage. METHODS: The mold concentrations in the dust in asthmatic children's bedrooms in water-damaged homes (N = 60) and control homes (N = 22) were measured by mold-specific quantitative polymerase chain reaction. RESULTS: Two molds, Scopulariopsis brevicaulis and Trichoderma viride, had significantly (P < 0.05) higher concentrations in asthmatics' homes compared with control homes and three other molds (Penicillium crustosum group, Stachybotrys chartarum, and Wallemia sebi) had P values <0.1. CONCLUSIONS: A relative moldiness index was developed to predict the likely development of asthma in water-damaged homes in Cleveland.

Asthma↗

Immunoglobulin G antibodies against environmental moulds in a Norwegian healthy population shows a bimodal distribution for Aspergillus versicolor.

Immunoglobulin G (IgG) antibodies against moulds related to indoor dampness problems are used as biomarkers to indicate exposure. In the present study, we evaluated the frequency of mould exposure in an adult healthy population by examining levels of mould-specific IgG antibodies in Norwegian blood donors. Using enzyme-linked immunosorbent assay, 106 blood donor sera were analyzed for IgG antibodies to Aspergillus versicolor, Penicillium chrysogenum, Cladosporium herbarum, Stachybotrys chartarum and Fusarium oxysporum. The levels of specific IgG antibodies to P. chrysogenum, C. herbarum and S. chartarum correlated (r = 0.46-0.62). Responses to A. versicolor were considerably stronger than to the other moulds, and another 996 blood donor sera were analyzed for IgG antibodies to this mould. Women had significantly higher levels of specific IgG antibodies to A. versicolor than men. The concentration of A. versicolor-specific IgG antibodies showed a non-Gaussian, bimodal distribution profile, in which 12.5% were defined as positive to exposure. This suggests that significant mould exposure in a healthy population can be calculated from mean + 1SD. Western blotting analyses showed that antibody responses to A. versicolor were largely directed against carbohydrate antigens of unknown saccharides.

Antibodies↗

Antifungal activity of thyme (Thymus vulgaris L.) essential oil and thymol against moulds from damp dwellings.

AIMS: To characterize antifungal activities of essential oil of thyme (Thymus vulgaris L.) and pure thymol, as comparative substance, on different mould species isolated from damp dwellings. METHODS AND RESULTS: Fifty samples of wall scrapes were collected from damp dwellings in Zagreb, the capital of Croatia. The members of the following mould genera were recovered from the samples: Aspergillus (44%), Penicillium (18%) Alternaria, Ulocladium, Absidia and Mucor (8%) Cladosporium, Trichoderma and Rhizopus (6%), and Chaetomium (2%). Two strains of Stachybotrys chartarum were isolated from damp dwellings in Slovakia. Antifungal activities of the thyme essential oil, which contains p-cymene (36.5%), thymol (33.0%) and 1,8-cineole (11.3%) as main components, and pure thymol were determined by the dilution method and exposure to vaporous phase of the oil. Minimum inhibitory concentrations (MIC) of both thymol and essential oil were bellow 20 microg ml(-1), except for Mucor spp. (50.20 microg ml(-1)). Thymol exhibited approximately three-times stronger inhibition than essential oil of thyme. The vaporous phase of the thyme essential oil (82 microg l(-1)) in glass chambers strongly suppressed the sporulation of moulds during 60 days of exposure. CONCLUSION: The thyme essential oil possesses a wide range spectrum of fungicidal activity. The vaporous phase of the oil exhibited long-lasting suppressive activity on moulds from damp dwellings. SIGNIFICANCE AND IMPACT OF THE STUDY: Essential oil of thyme and thymol could be used for disinfection of mouldy walls in the dwellings in low concentration.

Antifungal Agents↗

The production and characterization of monoclonal antibodies to the fungus Aspergillus versicolor.

UNLABELLED: Fungal exposure measurements in indoor environments require accurate and precise monitoring methods. Such techniques may be based on monoclonal antibodies (Mabs) and enzyme-linked immunosorbent assays (ELISA) and here we report the cross-reactivity patterns of Mabs produced against Aspergillus versicolor. Balb/c mice were immunized with the particulate fraction of homogenized spores and 46 Mabs (35 IgM, nine IgG3, two IgG1) were produced and tested for cross-reactivity against 55 fungal species. None of the Mabs was found to be species-specific for A. versicolor. Several Mabs strongly cross-reacted with most Aspergillus, Penicillium and Eurotium species and some Mabs also cross-reacted with Paecilomyces variotii and several Cladosporium and Stachybotrys species. Our results show that antibody responses in mice against spores of A. versicolor are dominated by highly cross-reactive antibodies of the IgM isotype. The widespread cross-reactivity suggests that the specificity of antibodies to be used for the detection of fungi in environmental samples need to be thoroughly characterized in order to avoid ambiguities in the interpretation of monitoring results. Furthermore, accurate estimates of spore concentrations may require the application of species-specific Mabs in order to avoid bias in result interpretation because of the differential reactivity of cross-reactive Mabs with different fungi. PRACTICAL IMPLICATIONS: Producers of monoclonal or polyclonal antibodies for the detection of fungi in environmental or clinical samples need to verify antibody reactivity patterns and accurately report that information to potential users. Furthermore, immunoassays based on mouse or human serum or purified immunoglobulin fractions need to consider antibody cross-reactivity as a potential confounding factor during interpretation of results.

Air Microbiology↗

Bacterial strains from moldy buildings are highly potent inducers of inflammatory and cytotoxic effects.

UNLABELLED: We aimed to identify inflammatory and cytotoxic potential of individual indoor air bacterial and fungal strains, as well as extracts of indoor air filter samples containing bacteria and fungi. Mouse RAW264.7 macrophages were exposed in vitro to four bacterial strains; Streptomyces californicus, Mycobacterium terrae, Bacillus cereus and Pseudomonas fluorescens, and three fungal strains; Penicillium spinulosum, Aspergillus versicolor and Stachybotrys chartarum. Furthermore, RAW264.7 macrophages were exposed to indoor air filter sample extracts representing 'low' (n = 21) and 'high' (n = 20) exposure to viable fungi or bacteria. Production of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) as well as cell viability were measured after 24 h exposure. The results show that the bacterial strains induce more profound production of NO, TNF-alpha and IL-6 than the studied fungal strains. They also decrease the viability of mouse macrophages. Similarly, the indoor air filter samples with high concentration of bacteria induced a statistically significant increase in TNF-alpha and IL-6 production as well as a decrease in cell viability. Altogether, these results suggest that indoor air bacterial strains are potent inducers of inflammatory responses and thus possibly related to adverse health effects of the inhabitants. PRACTICAL IMPLICATIONS: There is abundant documentation of the association between building dampness and mold and adverse health effects on occupants, but the causal agents of the effects are still unclear. In order to reveal these causal links, experimental studies with in vitro and in vivo methods are needed. The present findings shed new light on studies of the microbial constituents of indoor air in moldy buildings responsible for adverse health effects. These results imply that bacteria should also be monitored in cases of suspected microbial contamination of indoor air.

Air Microbiology↗

12,13-Epoxy-delta 9-trichothecenes as the probable mycotoxins responsible for stachybotryotoxicosis.

Stachybotrys atra cultures grown on oats produced five compounds toxic to brine shrimp; three are the sesquiterpenoid mycotoxins known as 12,13-epoxy-Delta(9)-trichothecenes. One trichothecene is roridin E, a known metabolite of Myrothecium verrucaria. The other two were hydrolyzed to verrucarol, the product of roridin and verrucarin hydrolysis. Spectroscopic data indicate that the two remaining compounds are also 12,13-epoxy-Delta(9)-trichothecenes. These metabolites are probably among those responsible for stachybotryotoxicosis.

Animals↗

Bacteria, molds, and toxins in water-damaged building materials.

Microbial toxins and eukaryotic cell toxicity from indoor building materials heavily colonized by fungi and bacteria were analyzed. The dominant colonizers at water-damaged sites of the building were Stachybotrys chartarum (10(3) to 10(5) visible conidia cm-2), Penicillium and Aspergillus species (10(4) CFU mg-1), gram-negative bacteria (10(4) CFU mg-1), and mycobacteria (10(3) CFU mg-1). The mycobacterial isolates were most similar to M. komossense, with 98% similarity of the complete 16S rDNA sequence. Limulus assay of water extracts prepared from a water-damaged gypsum liner revealed high contents of gram-negative endotoxin (17 ng mg-1 of E. coli lipopolysaccharide equivalents) and beta-D-glucan (210 ng mg-1 of curdlan equivalents). High-performance liquid chromatography analysis of the methanol extracts showed that the water-damaged gypsum liner also contained satratoxin (17 ng mg-1). This methanol-extracted substance was 200 times more toxic to rabbit skin and fetus feline lung cells than extract of gypsum liner sampled from a non-water-damaged site. The same extract contained toxin(s) that paralyzed the motility of boar spermatozoa at extremely low concentrations; the 50% effective concentration was 0.3 microgram of dry solids per ml. This toxicity was not explainable by the amount of bacterial endotoxin, beta-D-glucan, or satratoxin present in the same extract. The novel in vitro toxicity test that utilized boar spermatozoa as described in this article is convenient to perform and reproducible and was a useful tool for detecting toxins of microbial origin toward eukaryotic cells not detectable in building materials by the other methods.

Animals↗

Profiles of airborne fungi in buildings and outdoor environments in the United States.

We examined 12,026 fungal air samples (9,619 indoor samples and 2,407 outdoor samples) from 1,717 buildings located across the United States; these samples were collected during indoor air quality investigations performed from 1996 to 1998. For all buildings, both indoor and outdoor air samples were collected with an Andersen N6 sampler. The culturable airborne fungal concentrations in indoor air were lower than those in outdoor air. The fungal levels were highest in the fall and summer and lowest in the winter and spring. Geographically, the highest fungal levels were found in the Southwest, Far West, and Southeast. The most common culturable airborne fungi, both indoors and outdoors and in all seasons and regions, were Cladosporium, Penicillium, nonsporulating fungi, and Aspergillus. Stachybotrys chartarum was identified in the indoor air in 6% of the buildings studied and in the outdoor air of 1% of the buildings studied. This study provides industrial hygienists, allergists, and other public health practitioners with comparative information on common culturable airborne fungi in the United States. This is the largest study of airborne indoor and outdoor fungal species and concentrations conducted with a standardized protocol to date.

Air Microbiology↗