Airborne bacteria and fungal spores in the indoor environment. A case study in Singapore

The concentration of airborne fungal spores and bacteria as related to room temperature, humidity and occupancy levels within a library building in Singapore was determined. Measurement of indoor air quality with respect to microorganisms is of particular importance in tropical environments due to the extensive use of air‐conditioning systems and the potential implications for human health. This study has revealed a number of interesting relationships between the concentrations of fungal spores and bacteria in relation to both environmental and human factors. The levels of fungal spores measured in the indoor environment were approximately fifty times lower than those measured outside, probably because of the lowered humidity caused by air‐conditioning in the indoor environment. The variation in fungal spore concentration in the outdoor environment is likely to be due to the diurnal periodicity of spore release and the response to environmental factors such as light temperature and humidity. The indoor concentration of fungal spores in air was not clearly correlated to concentrations measured in air outside of the library building and remained relatively constant, unaffected by the difference in the numbers of occupants in the library. In contrast, the indoor concentrations of bacteria in air were approximately ten times higher than those measured outdoors, indicating a signficant internal source of bacteria. The elevated levels of indoor bacteria were primarily attributed to the number of library occupants. Increased human shedding of skin cells, ejection of microorganisms and particulates from the respiratory tract, and the transport of bacteria on suspended dust particles from floor surfaces probably accounts for the strong positive correlation between occupancy levels and the concentration of bacteria in internal air.     

A multi-laboratory comparative study of spore trap analyses

Fungal spore trap analyses currently are being marketed to the medical and environmental industries as a means of evaluating fungal bioaerosols. No studies comparing the results of these analyses have been conducted among laboratories providing these services. In the current study we compared the results from seven such laboratories with four different commercial spore trap cassettes with samples from four environmental conditions. The conditions included indoor air from a single location in a building under low, moderate and high agitation, and a sample from outside the same building. The means, ranges and standard deviations of total spore counts per cubic meter were respectively: low agitation indoor 514, 40-1933, 395; moderate agitation indoor 446, 80-1120, 290; high agitation indoor, 5154, 1510-15278, 3335; and outdoor 16012, 3700-28959, 6600. Results were similarly variable for the 27 spore categories that contribute to the total count. No consistent difference was observed in the precision of the kinds of spore traps. We concluded that spore trap analyses should be used with caution and should not be used as a sole method of assessing fungal spore populations and that standardized methods of analysis must be developed that include information about analytical precision of the sample data.     

Airborne fungal spores in 80 homes in the Latrobe Valley, Australia: levels, seasonality and indoor-outdoor relationship

Airborne viable and total fungal spores were sampled inside and outside 80 houses in the Latrobe Valley, Victoria, Australia as part of a larger indoor environmental study. Each residence was visited six times over a period of 1 year for sample collection, and fungal spore samples were collected from at least three indoor sites and from an outdoor site. Viable spores were sampled using an Andersen sampler, while total spores were assessed using a Burkard spore trap. Identification of fungal colonies to genera level was performed in two seasons; winter and late spring. The most common fungal genera/groups wereCladosporium, Penicillium, and yeasts, both indoors and outdoors in winter and late spring. Outdoor levels were higher than those indoors throughout the year, and a significant seasonal variation in spore levels was seen both indoors and outdoors with overall maxima in summer. Contrary to this trend, the levels ofAspergillus, yeasts,Cephalosporium andGliocladium were higher in winter. Most fungal genera were found in greater concentrations outdoors compared to indoors, butPenicillium was more common indoors. Outdoor spore levels were a significant influence on indoor levels, but seasonal differences suggest that other influences are important.     

Incidence and remediation of fungi in a sick building in Malaysia: a case study

Problems caused by indoor microbial, especially fungal growth, have been further exaggerated by the increased incidence of water intrusions, condensation from air-conditioning system, and other factors. While fungal contamination in a building can be one of the indicators of indoor air quality (IAQ), air quality assessment and remediation should be considered as important and should be carried out systematically. This study reports the incidence and remediation of an excessive fungal growth in a building in Universiti Sains Malaysia (USM), Pulau Pinang, Malaysia. Acting on an official report by the building owner and occupants, an immediate walk-through investigation was carried out between June and December 2009. A thorough sampling comprising swab and spore impactor techniques showed that the colony-forming unit per cubic meter of air (CFU/m³) levels were far above those in the guidelines of most developed countries. Thus, the building was declared a ‘sick building’ and closed to the publics. However, through immediate action with recommended methods to overcome the problem, the premise has been pronounced safe. This is a good example of successful teamwork that involved a continuous investigation by a team of experts along with comprehensive action taken by the occupants and other authorities responsible for the building.     

Do indoor plants contribute to the aeromycota in city buildings?

Many studies have focused on the sources of fungal contamination in indoor spaces. Pathogenic fungi have been detected in the potting mix of indoor plants; however, it is unclear if plants in indoor work spaces make qualitative or quantitative contributions to the aeromycota within buildings. The current work represents a field study to determine, under realistic office conditions, whether indoor plants make a contribution to the airborne aeromycota. Fifty-five offices, within two buildings in Sydney’s central business district, were studied over two seasonal periods: autumn and spring. We found that indoor plant presence made no significant difference to either indoor mould spore counts or their species composition. No seasonal differences occurred between autumn and spring samples. Indoor spore loads were significantly lower than outdoor levels, demonstrating the efficiency of the heating, ventilation and air conditioning systems in the buildings sampled. Neither the number of plants nor the species of plant used had an influence on spore loads; however, variations of those two variables offer potential for further studies. We conclude that conservative numbers of indoor plants make no substantial contribution to building occupants exposure to fungi.     

Rapid method for the determination of ochratoxin A in urine by immunoaffinity column clean-up and high-performance liquid chromatography

Buildings with poor indoor air quality (IAQ) frequently have many areas with surface fungal contamination. Studies have demonstrated that certain fungal genera (e.g., Cladosporium, Penicillium, and Stachybotrys) are able to grow on building materials such as wallpaper, drywall, and ceiling tiles, particularly after water damage has occurred. Due to the increasing awareness of sick building syndrome (SBS), it has become essential to identify building materials that prevent the interior growth of fungi. The objective of this study was to identify building materials that would not support the growth of certain fungal genera, regardless of whether an external food source was made available. The growth of three fungal genera (Cladosporium, Penicillium, and Stachybotrys) was evaluated on cellulose-containing ceiling tile (CCT) and inorganic ceiling tile (ICT). Both types of ceiling tile were exposed to environmental conditions which can occur inside a building. Our results show that ICT did not support the growth of these three fungal genera while CCT did. Our data demonstrate that ICT could serve as an ideal replacement for CCT. This revised version was published online in June 2006 with corrections to the Cover Date.     

Fungal spores are an important component of library air

The airborne fungal spore types were studied in different libraries in Delhi, using an Andersen sampler and a Burkard personal sampler, for culturable and non-culturable fungi respectively. The concentration inside the libraries, before and after the agitation of books, were compared with outside air. The major contributors to the library air areCladosporium, aspergilli/penicillia, smuts andAlternaria, varying from 50 to 14%. Some fungi (Cladosporium, Alternaria, smut,Penicillium chrysogenum andnigricans) showed seasonal occurrence, corresponding to their occurrence in the extramural environment. Aspergilli/penicillia,Drechslera, Curvularia andAspergillus flavus had a significantly higher concentration (P<0.01) inside the library, and recorded a significant increase in concentration after agitation of books. Air-conditioned libraries have low fungal spore concentrations, as compared to naturally ventilated libraries.     

Microbial contamination of storerooms at the Auschwitz-Birkenau Museum

Biodeterioration of heritage collections caused by microorganisms is a worldwide problem. To avoid degradation caused by biological contaminants transported into the indoor environment by air, proper bioaerosol protection is required. The aim of this study was to assess the level of microbial contamination of the Auschwitz-Birkenau Museum collection based on qualitative and quantitative analyses of bacteria and fungi isolated from the atmosphere and settled dust of museum storerooms. The obtained results demonstrated that a correctly operated air-conditioning system and limiting the number of visitors in the studied storerooms can significantly inhibit microbial contamination of the air and decrease deposition of bacterial and fungal particulates on exhibit surfaces. The performed analyses confirm that an aerobiological assessment of museum premises is a useful tool in their hygienic evaluation and, if necessary, in decision-making regarding interventions to minimize biological decay of collections.     

An assessment of the air quality in indoor and outdoor air with reference to fungal spores and pollen grains in four working environments in Kerala, India

Qualitative and quantitative analyses of airborne fungal spores and pollen grains in four working environments (market, saw mill, poultry and cow sheds) in Thiruvananthapuram, the capital city of Kerala, India, were carried out for 2 years using Burkard Personal Slide Sampler and Andersen Two-Stage Sampler. Total spore concentration in these sites was always higher in indoor environments than in outdoor environments. Difference in concentration was not statistically significant in any of these work places except in saw mill (t test, p < 0.05). The highest spore concentration was recorded here followed by market, poultry and cow sheds. A total of 32 fungal spore types from indoor environments and 33 spore types from outdoor environments were recorded. Of them, 16 spore types were common to all the sites. Ameropsores, Cladosporium, other basidiospores, Ganoderma and Nigrospora were the dominant spore types in both indoor and outdoor environments. A total of 27 species of viable fungi from indoor and 24 species from outdoor environments were identified. Penicillium citrinum, Aspergillus flavus and Aspergillus niger were the most dominant viable fungi isolated. In contrast, total pollen concentration was always higher in outdoor environments than in indoor environments. Twenty-nine pollen types from indoor and 32 pollen types from outdoor were captured during the sampling. Poaceae, Cocos, Artocarpus, Amaranthus/Chenopodium and Tridax were the common and dominant pollen types observed in all the sites. Peak spore and pollen incidence were recorded during the late rainy and dry seasons (October–February) in both indoor and outdoor environments. The study revealed high prevalence of predominantly allergenic fungal spores and pollen grains in all the four work places. Workers/visitors are at potential risk of susceptibility to respiratory/allergic disorders.     

Characterization of an airborne microbial community: A case study in the archive of the University of Coimbra,Portugal

Understanding the structure of indoor airborne microbial communities could be useful in optimizing conservation and disinfection procedures in archive repositories, preventing the biodeterioration of stored collections. In this study we characterized the microbial air community inside the Archive of the University of Coimbra, by identifying different fungal and bacterial organisms retrieved from air samples. The microbial contamination was determined using conventional culture methods, and the isolates were typified using morphological techniques. Results indicated a low microbial air contamination (107 ± 12 CFU/m³), particularly regarding fungal propagules (6 ± 1 CFU/m³). Fungal isolates were identified using ITS-DNA sequencing. Among fungal isolates, Penicillium was the most frequent genus, and Penicillium griseofulvum was the predominant species. Simpson diversity index (1-D) was applied to phenotypic and genotypic results. Total phenotypic diversity varied from 0.4 to 0.8 and regarding fungal species, the diversity was higher than 0.5. These results were compared with previous analyses of the Archive's air, suggesting that short-term changes in atmospheric conditions may influence the indoor air microbial community structure.     

Determining Fungi rRNA Copy Number by PCR

The goal of this project is to improve the quantification of indoor fungal pollutants via the specific application of quantitative PCR (qPCR). Improvement will be made in the controls used in current qPCR applications. This work focuses on the use of two separate controls within a standard qPCR reaction. The first control developed was the internal standard control gene, benA. This gene encodes for β-tubulin and was selected based on its single-copy nature. The second control developed was the standard control plasmid, which contained a fragment of the ribosomal RNA (rRNA) gene and produced a specific PCR product. The results confirm the multicopy nature of the rRNA region in several filamentous fungi and show that we can quantify fungi of unknown genome size over a range of spore extractions by inclusion of these two standard controls. Advances in qPCR have led to extremely sensitive and quantitative methods for single-copy genes; however, it has not been well established that the rRNA can be used to quantitate fungal contamination. We report on the use of qPCR, combined with two controls, to identify and quantify indoor fungal contaminants with a greater degree of confidence than has been achieved previously. Advances in indoor environmental health have demonstrated that contamination of the built environment by the filamentous fungi has adverse impacts on the health of building occupants. This study meets the need for more accurate and reliable methods for fungal identification and quantitation in the indoor environment.     

Long-term use of high-efficiency vacuum cleaners and residential airborne fungal-spore exposure

Exposure to fungal allergens is an important contributor to allergic respiratory disease, but information on the efficacy of residential fungal allergen-avoidance in allergic-disease management is lacking. Using vacuum cleaners with high-efficiency exhaust filtration is one method recommended for reducing residential allergen exposure levels, but their use to reduce fungal-spore exposure levels has not been evaluated. To evaluate the effectiveness of high-efficiency vacuuming to control airborne fungal-spore levels, fungal bioaerosols were repeatedly assessed over the course of 10 months in homes randomly assigned to groups using either conventionally filtered (control) or high-efficiency-filtered vacuum cleaners for routine vacuum cleaning. Air samples were analyzed for three fungal-spore categories representing taxa with predominantly outdoor sources and one representing taxa that commonly have indoor sources. In a two-way analysis of variance, sampling period had a significant effect on mean levels of all fungal-spore categories. Vacuum cleaner type had a marginally significant effect on the indoor spore category, with one high-efficiency vacuum group mean (of three) significantly lower than one control mean. No effect was observed of vacuum cleaner type on outdoor spore categories. Including home-environment variables in analysis of covariance models strengthened the effect of the vacuum-type treatment on the indoor spore category, with no effect on the three outdoor spore categories. Decreased indoor spore levels vs. controls were only observed in high-efficiency vacuum groups during the last sampling period, at the end of the heating season. The results suggest that using a vacuum with high-efficiency filtered exhaust could have some modest effectiveness in controlling airborne fungal-spore exposure in homes when infiltration of outdoor air is very limited.     

Fungal and Bacterial Communities in Indoor Dust Follow Different Environmental Determinants

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

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

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

Fungal population in the atmosphere of Ismailia City

Fungal spore populations in the outdoor and indoor atmosphere of Ismailia have been studied during the period from March 1992 to May 1993. A total of 23 350 cfu and 73 species were recorded.Cladosporium cladosporioides, Aureobasidium pullulans andAspergillus flavus were the most abundant. The indoor and outdoor mycoflora showed marked quantitative and qualitative differences. In view of count, recorded species could be categorized into three groups as follows: (a) species showing higher counts in out-than indoor, (b) species showing the opposite trend i.e. lower counts in out-door than indoor, (c) species showing approximately equal counts in out- and indoor. Regarding seasonal periodicity, March and either September or October showed the highest count for both normal fungal flora (NFF) and opportunistic fungal flora (OFF). While January and July showed the lowest count of them both, May but not July was the lowest as for outdoor NFF.     

Long-term use of high-efficiency vacuum cleaners and residential airborne fungal-spore exposure

Exposure to fungal allergens is an important contributor to allergic respiratory disease, but information on the efficacy of residential fungal allergen-avoidance in allergic-disease management is lacking. Using vacuum cleaners with high-efficiency exhaust filtration is one method recommended for reducing residential allergen exposure levels, but their use to reduce fungal-spore exposure levels has not been evaluated. To evaluate the effectiveness of high-efficiency vacuuming to control airborne fungal-spore levels, fungal bioaerosols were repeatedly assessed over the course of 10 months in homes randomly assigned to groups using either conventionally filtered (control) or high-efficiency-filtered vacuum cleaners for routine vacuum cleaning. Air samples were analyzed for three fungal-spore categories representing taxa with predominantly outdoor sources and one representing taxa that commonly have indoor sources. In a two-way analysis of variance, sampling period had a significant effect on mean levels of all fungal-spore categories. Vacuum cleaner type had a marginally significant effect on the indoor spore category, with one high-efficiency vacuum group mean (of three) significantly lower than one control mean. No effect was observed of vacuum cleaner type on outdoor spore categories. Including home-environment variables in analysis of covariance models strengthened the effect of the vacuum-type treatment on the indoor spore category, with no effect on the three outdoor spore categories. Decreased indoor spore levels vs. controls were only observed in high-efficiency vacuum groups during the last sampling period, at the end of the heating season. The results suggest that using a vacuum with high-efficiency filtered exhaust could have some modest effectiveness in controlling airborne fungal-spore exposure in homes when infiltration of outdoor air is very limited.     

Airborne fungal spores in a sawmill environment in Palakkad District, Kerala, India

Concentration of airborne fungal spores inindoor and outdoor environments of a sawmill in Palakkad district of Kerala, India was studied with Burkard Personal Slide Sampler from January to December 1997. Total spore concentration in the indoor and outdoor showed a 3:2 ratio. Higher spore count was observed in indoor in January and in outdoor in October. Thirty three fungal spore types were identified from the indoor and twenty six from the outdoor. Aspergillus/Penicillium, Cladosporium, Nigrospora, Ganoderma, `other basidiospores' and ascospores were the dominant components of the airspora. Aspergillus/Penicillium, the most dominant spore type in the indoor contributed 51.19% and Cladosporium, the most dominant spore type in the outdoor contributed 44.75% of the total spores. The study revealed high prevalence of predominantly allergenic fungal spores in the sawmill environment.     

Fungal population in the atmosphere of Ismailia City

Fungal spore populations in the outdoor and indoor atmosphere of Ismailia have been studied during the period from March 1992 to May 1993. A total of 23 350 cfu and 73 species were recorded,Cladosporium cladosporioides, Aureobasidium pullulans andAspergillus flavus were the most abundant. The indoor and outdoor mycoflora showed marked quantitative and qualitative differences. In view of count, recorded species could be categorized into three groups as follows: (a) species showing higher counts in out- than indoor, (b) species showing the opposite trend i.e. lower counts in out-door than indoor, (c) species showing approximately equal counts in out- and indoor. Regarding seasonal periodicity, March and either September or October showed the highest count for both normal fungal flora (NFF) and opportunistic fungal flora (OFF). While January and July showed the lowest count of them both, May but not July was the lowest as for outdoor NFF.     

Possible cause of allergy for the librarians: books manipulation and ventilation as sources of fungus spores spreading

The objective of this study was to investigate the airborne viable spore concentrations and identify the fungal species in all indoor spaces from the lending library at the Technical University “Gheorghe Asachi” Iaşi, Romania. Samples were collected using the settle plate method and swab samples from PC cooler fan grids as well as from the wall in it’s vicinity and from paper/wood fragments. There were no air conditioning systems in the library rooms. The heating systems were standard with an environmental temperature of 20°C in winter, except for the storage area of old/rare books stacks II, where the temperature was below 15°C and the humidity was very high due to water infiltrations in the walls and poor maintenance. More than 296 fungal colonies from over 78 samples were identified, enumerated, and reported. Indoor airborne fungal spore deposition rates were within the range of 419–1,677 CFU/m², with the predominance of genera being Aspergillus spp., Penicillium spp., Cladosporium spp., Alternaria spp. and Chaetomium spp. Approximately ten fungal colonies could not be identified. The PC fans move particles from the low levels (floor) to the air, and are thus responsible for maintaining a constant air velocity and contribute to fungal-spore aerosolization, transport, deposition and resuspension. Book paper and wood furniture are known to be suitable substrates for cellulose degrading fungi.     

Airborne fungi in four stations of the St. Petersburg Underground railway system

The fungal and bacterial aerobiota of four St. Petersburg Underground stations has been examined over a 4-month period. In the indoor air of St. Petersburg Underground 50 fungal species were found, among which were likely deteriogenic fungi. The most prevailing genera were Acremonium, Aspergillus, Cladosporium and Penicillium. Fungal spore density in the underground air was within the sanitary level accepted for public buildings. The spore densities and specificities correlated with the station type. A more specific (independent of outdoor) air mycobiota was found in deeper stations. All fungal isolates were tested in laboratory conditions for their ability to produce extracellular proteinase, phospholipase, and hemolytic activities which can be associated to virulence. Only 2 of the 75 isolates expressed a high level of all three activities. Assuming this figure can serve as a rough assessment of pathogenicity potential, the risk of invasive mycoses was not considered significant. But taking into account the situation with peak-hours overcrowding, it may be concluded that the risk of “mould” allergic diseases for some categories of the underground passengers in St. Petersburg does exist.