Co-occurrence of toxic bacterial and fungal secondary metabolites in moisture-damaged indoor environments

Toxic microbial secondary metabolites have been proposed to be related to adverse health effects observed in moisture-damaged buildings. Initial steps in assessing the actual risk include the characterization of the exposure. In our study, we applied a multi-analyte tandem mass spectrometry-based methodology on sample materials of severely moisture-damaged homes, aiming to qualitatively and quantitatively describe the variety of microbial metabolites occurring in building materials and different dust sample types. From 69 indoor samples, all were positive for at least one of the 186 analytes targeted and as many as 33 different microbial metabolites were found. For the first time, the presence of toxic bacterial metabolites and their co-occurrence with mycotoxins were shown for indoor samples. The bacterial compounds monactin, nonactin, staurosporin and valinomycin were exclusively detected in building materials from moist structures, while chloramphenicol was particularly prevalent in house dusts, including settled airborne dust. These bacterial metabolites are highly bioactive compounds produced by Streptomyces spp., a group of microbes that is considered a moisture damage indicator in indoor environments. We show that toxic bacterial metabolites need to be considered as being part of very complex and diverse microbial exposures in 'moldy' buildings. PRACTICAL IMPLICATIONS: Bacterial toxins co-occur with mycotoxins in moisture-damaged indoor environments. These compounds are measurable also in settled airborne dust, indicating that inhalation exposure takes place. In attempts to characterize exposures to microbial metabolites not only mycotoxins but also bacterial metabolites have to be targeted by the analytical methods applied. We recommend including analysis of samples of outdoor air in the course of future indoor assessments, in an effort to better understand the outdoor contribution to the indoor presence of microbial toxins. There is a need for a sound risk assessment concerning the exposure to indoor microbial toxins at concentrations detectable in moisture-damaged indoor environments.     

Molds and mycotoxins in dust from water‐damaged homes in New Orleans after hurricane Katrina

Abstract Abstract Dust collected in New Orleans homes mold‐contaminated because of the flooding after hurricane Katrina was analyzed for molds and mycotoxins. The mycoflora was studied by cultivation and quantitative PCR for selected molds. The most commonly found mold taxa were Aspergillus, Cladosporium, and Penicillium. Verrucarol, a hydrolysis product of macrocyclic trichothecenes predominately produced by Stachybotrys spp. was identified in three dust samples by gas chromatography‐tandem mass spectrometry, and sterigmatocystin (produced by various Aspergillus spp.) was found in two samples by high pressure liquid chromatography‐tandem mass spectrometry. This is the first demonstration of mycotoxins in Katrina‐associated dust samples. The analytical methods used represent valuable tools in further studies on bioaerosol exposure and health risks.     

Coupled simulation of heat and moisture transport in air and porous materials for the assessment of moisture related damage

This paper describes the coupling of a model for heat and moisture transport in porous materials to a commercial Computational Fluid Dynamics (CFD) package. The combination of CFD and the material model makes it possible to assess the risk of moisture related damage in valuable objects for cases with large temperature or humidity gradients in the air. To couple both models the choice was made to integrate the porous material model into the CFD package. This requires the heat and moisture transport equations in the air and the porous material to be written down in function of the same transported variables. Validation with benchmark experiments proved the good functionality of the coupled model. A simulation study of a microclimate vitrine for paintings shows that phenomena observed in these vitrines are well predicted by the model and that data generated by the model provides additional insights in the physical mechanisms behind these phenomena.     

Higher measured moisture in California homes with qualitative evidence of dampness

Relationships between measured moisture and qualitative dampness indicators (mold odor, visible mold, visible water damage, or peeling paint) were evaluated using data collected from California homes in a prospective birth cohort study when the infants were 6 or 12 months of age (737 home visits). For repeated visits, agreement between observation of the presence/absence of each qualitative indicator at both visits was high (71–87%, P < 0.0001). Among individual indicators, musty odor and visible mold were most strongly correlated with elevated moisture readings. Measured moisture differed significantly between repeated visits in opposite seasons (P < 0.0001), and dampness increased with the number of indicators in a home. Linear mixed‐effect models showed that 10‐unit increases in maximum measured moisture were associated with the presence of 0.5 additional dampness indicators (P < 0.001). Bedroom (BR) walls were damper than living room (LR) walls in the same homes (P < 0.0001), although both average and maximum readings were positively correlated across room type (r = 0.75 and 0.67, respectively, both P < 0.0001). Exterior walls were significantly damper than interior walls (P < 0.0001 in both LRs and BRs), but no differences were observed between maximum wall readings and measurements at either window corners or sites of suspected dampness.     

Levels and sources of volatile organic compounds in homes of children with asthma

Many volatile organic compounds (VOCs) are classified as known or possible carcinogens, irritants, and toxicants, and VOC exposure has been associated with the onset and exacerbation of asthma. This study characterizes VOC levels in 126 homes of children with asthma in Detroit, Michigan, USA. The total target VOC concentration ranged from 14 to 2274 μg/m³ (mean = 150 μg/m³; median = 91 μg/m³); 56 VOCs were quantified; and d‐limonene, toluene, p, m‐xylene, and ethyl acetate had the highest concentrations. Based on the potential for adverse health effects, priority VOCs included naphthalene, benzene, 1,4‐dichlorobenzene, isopropylbenzene, ethylbenzene, styrene, chloroform, 1,2‐dichloroethane, tetrachloroethene, and trichloroethylene. Concentrations varied mostly due to between‐residence and seasonal variation. Identified emission sources included cigarette smoking, solvent‐related emissions, renovations, household products, and pesticides. The effect of nearby traffic on indoor VOC levels was not distinguished. While concentrations in the Detroit homes were lower than levels found in other North American studies, many homes had elevated VOC levels, including compounds that are known health hazards. Thus, the identification and control of VOC sources are important and prudent, especially for vulnerable individuals. Actions and policies to reduce VOC exposures, for example, sales restrictions, improved product labeling, and consumer education, are recommended.     

Moisture damage in home associates with systemic inflammation in children

This study investigated the association between confirmed moisture damage in homes and systemic subclinical inflammation in children. Home inspections were performed in homes of 291 children at the age of 6 years. Subclinical inflammation at the age of 6 years was assessed by measuring the circulating levels of C‐reactive protein (CRP) and leukocytes in peripheral blood and fractional exhaled nitric oxide (FeNO). Proinflammatory cytokines interleukin (IL)‐1β and IL‐6 and tumor necrosis factor (TNF)‐α were measured in unstimulated, and in phorbol 12‐myristate 13‐acetate and ionomycin (PI), lipopolysaccharide (LPS), or peptidoglycan (PPG)‐stimulated whole blood. Major moisture damage in the child's main living areas (living room, kitchen, or child's bedroom) and moisture damage with mold in the bathroom were associated with increased levels of CRP and stimulated production of several proinflammatory cytokines. There were no significant associations between moisture damage/visible mold and leukocyte or FeNO values. The results suggest that moisture damage or mold in home may be associated with increased systemic subclinical inflammation and proinflammatory cytokine responsiveness.     

Children's homes--determinants of moisture damage and asthma in Finnish residences

Certain housing characteristics increase the risk for moisture damage, which has been associated with increased risk for asthma in children. Modeling moisture damage as a function of these characteristics could therefore provide a simple tool to estimate building-related risk for asthma. This study aimed to find out specific associations between asthma case-control status of children and moisture damage and housing characteristics. The data consisted of information on 121 asthmatic children and predominately two age-, gender- and place of residence-matched control children for every case, and information on moisture damage and housing characteristics in the homes of the children. In a previous study, we found a statistically significant association between moisture damage observations in main living areas and asthma in children. Using logistic regression, five models were formulated to predict moisture damage status of the homes and moisture damage status of living areas. The models were able to classify the damage status correctly in 65.0-87.7% of the homes (kappa values 0.10-0.47) as functions of housing characteristics. None of the models qualified as a significant determinant of the case-control status of the children. PRACTICAL IMPLICATIONS: It can be hypothesized that building-related risk for asthma could be roughly estimated using models predicting moisture damage status of buildings as a function of easily obtainable housing characteristics. The results of this study indicated that, with a moderate certainty, it is possible to model moisture damage status of buildings using housing characteristics. However, the models developed did not associate with asthma in children. In conclusion, it was not possible to estimate the risk for asthma by studying housing characteristics only, but detailed information on moisture damage (e.g. location of damage) was crucial for such estimation.     

Microbial specialists in below‐grade foundation walls in Scandinavia

Below‐grade foundation walls are often exposed to excessive moisture by water infiltration, condensation, leakage, or lack of ventilation. Microbial growth in these structures depends largely on environmental factors, elapsed time, and the type of building materials and construction setup. The ecological preferences of Actinomycetes (Actinobacteria) and the molds Ascotricha chartarum, Myxotrichum chartarum (Ascomycota), Geomyces pannorum, and Monocillium sp. (Hyphomycetes) have been addressed based on analyses of 1764 samples collected in below‐grade spaces during the period of 2001–2012. Our results show a significant correlation between these taxa and moist foundation walls as ecological niches. Substrate preference was the strongest predictor of taxa distribution within the wall, but the taxa's physiological needs, together with gradients of abiotic factors within the wall structure, also played a role. Our study describes for the first time how the wall environment affects microbial growth.     

Next‐generation DNA sequencing reveals that low fungal diversity in house dust is associated with childhood asthma development

Dampness and visible mold in homes are associated with asthma development, but causal mechanisms remain unclear. The goal of this research was to explore associations among measured dampness, fungal exposure, and childhood asthma development without the bias of culture‐based microbial analysis. In the low‐income, Latino CHAMACOS birth cohort, house dust was collected at age 12 months, and asthma status was determined at age 7 years. The current analysis included 13 asthma cases and 28 controls. Next‐generation DNA sequencing methods quantified fungal taxa and diversity. Lower fungal diversity (number of fungal operational taxonomic units) was significantly associated with increased risk of asthma development: unadjusted odds ratio (OR) 4.80 (95% confidence interval (CI) 1.04–22.1). Control for potential confounders strengthened this relationship. Decreased diversity within the genus Cryptococcus was significantly associated with increased asthma risk (OR 21.0, 95% CI 2.16–204). No fungal taxon (species, genus, class) was significantly positively associated with asthma development, and one was significantly negatively associated. Elevated moisture was associated with increased fungal diversity, and moisture/mold indicators were associated with four fungal taxa. Next‐generation DNA sequencing provided comprehensive estimates of fungal identity and diversity, demonstrating significant associations between low fungal diversity and childhood asthma development in this community.     

Indoor air microbes and respiratory symptoms of children in moisture damaged and reference schools

Microbial indoor air quality and respiratory symptoms of children were studied in 24 schools with visible moisture and mold problems, and in eight non-damaged schools. School buildings of concrete/brick and wooden construction were included. The indoor environment investigations included technical building inspections for visible moisture signs and microbial sampling using six-stage impactor for viable airborne microbes. Children's health information was collected by questionnaires. The effect of moisture damage on concentrations of fungi was clearly seen in buildings of concrete/brick construction, but not in wooden school buildings. Occurrence of Cladosporium, Aspergillus versicolor, Stachybotrys, and actinobacteria showed some indicator value for moisture damage. Presence of moisture damage in school buildings was a significant risk factor for respiratory symptoms in schoolchildren. Association between moisture damage and respiratory symptoms of children was significant for buildings of concrete/brick construction but not for wooden school buildings. The highest symptom prevalence was found during spring seasons, after a long exposure period in damaged schools. The results emphasize the importance of the building frame as a determinant of exposure and symptoms.     

The effect of ozonization on furniture dust: Microbial content and immunotoxicity in vitro

Moisture and mold problems in buildings contaminate also the furniture and other movable property. If cleaning of the contaminated furniture is neglected, it may continue to cause problems to the occupants even after the moisture-damage repairs. The aim of this study was to determine the effectiveness of high-efficiency ozone treatment in cleaning of the furniture from moisture-damaged buildings. In addition, the effectiveness of two cleaning methods was compared. Samples were vacuumed from the padded areas before and after the treatment. The microbial flora and concentrations in the dust sample were determined by quantitative cultivation and QPCR-methods. The immunotoxic potential of the dust samples was analyzed by measuring effects on cell viability and production of inflammatory mediators in vitro. Concentrations of viable microbes decreased significantly in most of the samples after cleaning. Cleaning with combined steam wash and ozonisation was more effective method than ozonising alone, but the difference was not statistically significant. Detection of fungal species with PCR showed a slight but nonsignificant decrease in concentrations after the cleaning. The immunotoxic potential of the collected dust decreased significantly in most of the samples. However, in a small subgroup of samples, increased concentrations of microbes and immunotoxicological activity were detected. This study shows that a transportable cleaning unit with high-efficiency ozonising is in most cases effective in decreasing the concentrations of viable microbes and immunotoxicological activity of the furniture dust. However, the method does not destroy or remove all fungal material present in the dust, as detected with QPCR analysis, and in some cases the cleaning procedure may increase the microbial concentrations and immunotoxicity of the dust.     

Salt damage to brick kiln walls in Japan: spatial and seasonal variation of efflorescence and moisture content

To clarify the mechanism of salt fretting of a brick building, monthly observations were made over one year at a two-storey brick kiln in central Japan. Products of weathering that had fallen were collected and weighed; salts were identified using XRD and the moisture content of the brick was measured at its surface using a portable infrared optical moisture meter. Gypsum (CaSO₄·2H₂O) was found at the bottom of the first floor wall in the warm and humid season. Thenardite (Na₂SO₄) is dominant on the first floor wall; it is abundant in the cold and dry season but sparse in the warm and humid season. Magnesium sulphate is dominant on the second floor wall; epsomite (MgSO₄·7H₂O) was observed in the dry-to-wet season and hexahydrite (MgSO₄·6H₂O) in the wet-to-dry season. Seasonal changes in salt abundance result from the temperature dependence of the solubilities and equilibrium relative humidities (ERH) of each salt. The amount of brick decay material due to salt weathering is much greater from the second floor wall than from the first floor wall and is greater from spring to the end of summer. These seasonal and spatial variations of the rate of brick decay can be explained by a combination of factors: (1) the types of salt, (2) efflorescence or sub-florescence and (3) salt deliquescence.     

Microbial growth in building material samples and occupants’ health in severely moisture‐damaged homes

There is no commonly approved approach to detect and quantify the health‐relevant microbial exposure in moisture‐damaged buildings. In 39 single‐family homes with severe moisture damage, we studied whether concentrations of viable microbes in building material samples are associated with health among 71 adults and 68 children, and assessed with symptoms questionnaires, exhaled NO, and peak expiratory flow (PEF) variability. Symptoms were grouped into three scores: upper respiratory symptoms, lower respiratory symptoms, and general symptoms. The homes were divided into three groups based on viable counts of fungi, actinomycetes, and total bacteria cultivated from building material samples. Highest group of actinomycete counts was associated with more general symptoms, worse perceived health, and higher daily PEF variability (aOR 12.51; 1.10‐141.90 as compared to the lowest group) among adults, and with an increase in lower respiratory symptoms in children, but the confidence intervals were wide. We observed significant associations of fungal counts and total microbial score with worse perceived health in adults. No associations with exhaled NO were observed.     

Fungi and their secondary metabolites in water-damaged indoors after a major flood event in eastern Croatia

In winter and summer of 2016 and 2017, airborne fungi and house dust were collected in indoors of the village Gunja, which had been flooded, and the control village Gornji Stupnik (Croatia) in order to explore variations of fungal indoor levels, particularly Aspergilli section Nidulantes series Versicolores, as well as fungal metabolites in dust. Levels of airborne Aspergilli (Versicolores) were three times as high in winter and summer in Gunja than in the control village, while dustborne isolates were equally present in both locations. Sequencing of the calmodulin gene region revealed that among Aspergilli (Versicolores), A. jensenii and A. creber were dominant and together with A. puulaauensis, A. tennesseensis and A. venenatus produced sterigmatocystin and 5-methoxysterigmatocystin (HPLC coupled with mass spectrometry); A. amoenus, A. fructus, A. griseoaurantiacus, A. pepii, and A. protuberus produced sterigmatocystin but not 5-methoxysterigmatocystin; A. sydowii did not produce any of these toxins. A total of 75 metabolites related to Penicillium (29), Aspergillus (22), Fusarium (10), Alternaria (5), Stachybotrys (2), and other fungi (7) were detected in dust by liquid chromatography-tandem mass spectrometry. The majority of metabolites including sterigmatocystin and 5-methoxysterigmatocystin exhibited a higher prevalence in winter in Gunja.     

Field and laboratory performance comparison for asphalt mixtures with different moisture conditioning systems

The purpose of this study is to compare laboratory and field performances for dense graded asphalt mixtures. Control road pavement section was constructed in Black Sea Coast Highway. Marshall identical control samples produced in laboratory and core samples taken from wearing courses before the traffic opened were used. Three different moisture conditioning methods were applied to control laboratory and core samples. Mechanical properties of samples were evaluated with indirect tension, indirect tension strength and repeated creep tests. Indirect tensile strength test results for laboratory mixtures were found 1.22 and 1.30 times more in accordance with field samples at 10 °C and 20 °C respectively. Marshall samples gave higher resilient modulus for all control and conditioned mixtures. Repeated creep test results also proved the difference between laboratory and field sample performance. Laboratory samples protected their structural integrity along with the test duration and did not showed tertiary creep. It is concluded from this study that laboratory samples state expressly higher performance according to the core field samples.     

Meta-analyses of the associations of respiratory health effects with dampness and mold in homes

The Institute of Medicine (IOM) of the National Academy of Sciences recently completed a critical review of the scientific literature pertaining to the association of indoor dampness and mold contamination with adverse health effects. In this paper, we report the results of quantitative meta-analyses of the studies reviewed in the IOM report plus other related studies. We developed point estimates and confidence intervals (CIs) of odds ratios (ORs) that summarize the association of several respiratory and asthma-related health outcomes with the presence of dampness and mold in homes. The ORs and CIs from the original studies were transformed to the log scale and random effect models were applied to the log ORs and their variance. Models accounted for the correlation between multiple results within the studies analyzed. Central estimates of ORs for the health outcomes ranged from 1.34 to 1.75. CIs (95%) excluded unity in nine of 10 instances, and in most cases the lower bound of the CI exceeded 1.2. Based on the results of the meta-analyses, building dampness and mold are associated with approximately 30-50% increases in a variety of respiratory and asthma-related health outcomes. PRACTICAL IMPLICATIONS: The results of these meta-analyses reinforce the IOM's recommendation that actions be taken to prevent and reduce building dampness problems, and also allow estimation of the magnitude of adverse public health impacts associated with failure to do so.     

Improved health after intervention in a school with moisture problems

In a school with floor moisture problems, the personnel had complaints consistent with the sick-building syndrome (SBS). Interventive measures including the laying of a ventilated floor were undertaken to eliminate the emissions. To examine if the intervention resulted in positive health effects, 34 personnel and 336 pupils were interviewed just before the intervention and also 7 months after. Also were interviewed 21 personnel and 224 pupils at an adjacent school serving as a control. Compared with the control school, the problem school showed more complaints, more general symptoms and more symptoms from the eyes, airways and skin, both among the personnel and the pupils. In the post-intervention examinations, the excess of symptoms among the personnel had almost disappeared. Among the pupils, the frequency of eye irritation was reduced but a general improvement of the other symptoms was not as obvious. However, after adjustment for a recent common cold, atopy and stress among the pupils, only one symptom ("stuffy nose") remained significantly elevated. In conclusion, the intervention was followed by positive health effects, supporting the hypothesis that emissions from building material had contributed to the excess of symptoms. A recent common cold was highly related to the symptoms and should be considered in future SBS studies.     

Slowly decreasing mucosal hyperreactivity years after working in a school with moisture problems

In our first study in 1995, teachers, who had worked in a water-damaged school for more than 5 years, were tested for nasal histamine reactivity by rhinostereometry. They were found to have significantly increased reactivity compared with teachers in a school without these indoor-climate problems. This finding could not be explained by differences in atopy or other personal characteristics. In this 2-year follow-up study (1995-97), 26 of 28 teachers in the target school and all 18 teachers in the control school, who participated in the initial study, accepted to take part. They were tested with the same histamine provocation procedure and answered the same questionnaire as 2 years earlier. Technical measurements of temperature, relative humidity, dust, carbon dioxide, formaldehyde and total volatile organic compounds (TVOC) were carried out in both schools during the time period between the two test occasions. In this provocation test, the teachers from the repaired water-damaged school still demonstrated an increased reactivity to histamine compared with the teachers in the control school, but the difference between the growth curves of the provocation tests was less than in 1995. Teachers in the target school still complained about the indoor air quality more than their colleagues, although the complaints were less common. No major differences were observed in the technical investigation between the two schools and the measurements were all within values usually seen in schools in northern countries. Our conclusion is that the observed nasal mucosal hyperreactivity among the teachers in the renovated water-damaged school seems to persist over years and only slowly decrease even after successful remedial measures have been taken.