Prevalence of fungi in carpeted floor environment: Analysis of dust samples from living-rooms,bedrooms, offices and school classrooms

The results of 100 carpet dust analyses from atopic individuals' environment were compared according to the sampling period or the location. Dust samples were collected with a standard domestic vacuum cleaner, in locations with carpeted floor: in residences (living-room and/or bedroom), in school classrooms and in offices. The quantities of fungi vary from 5000 CFU/g to 66 000 000 CFU/g of dust. More than 100 species were isolated by dilution plating. The main species found in carpet dust wereEurotium repens, Penicillium chrysogenum, Alternaria alternata, Aureobasidium pullulans andPhoma herbarum. Strict xerophilic species were rather rare and detected in small quantities. Differences in the distribution of the CFU concentrations were examined for the four different sampling locations and were statistically significant (P=0.0174). In this study, schools were open spaces, and offices, mostly with air conditioning systems, were locations in which air is not confined. This, added to frequent professional carpet cleaning, probably explains the lowest levels of fungal concentration found in these locations. The majority of the homes had the largest fungal concentration in the living-room (median: 2×10⁵ CFU/g) while some bedrooms (median: 7×10⁴ CFU/g) had the highest concentrations. It is suggested that, when fungi are suspected to be the origin of respiratory allergy or irritating symptoms, the mycoflora of the bedroom, principally, should be investigated first.     

Prevalence of fungi in carpeted floor environment: analysis of dust samples from living-rooms,bedrooms, offices and school classrooms

The results of 100 carpet dust analyses from atopic individuals' environment were compared according to the sampling period or the location. Dust samples were collected with a standard domestic vacuum cleaner, in locations with carpeted floor: in residences (living-room and/or bedroom), in school classrooms and in offices. The quantities of fungi vary from 5000 CFU/g to 66 000 000 CFU/g of dust. More than 100 species were isolated by dilution plating. The main species found in carpet dust wereEurotium repens, Penicillium chrysogenum, Alternaria alternata, Aureobasidium pullulans andPhoma herbarum. Strict xerophilic species were rather rare and detected in small quantities. Differences in the distribution of the CFU concentrations were examined for the four different sampling locations and were statistically significant (P=0.0174). In this study, schools were open spaces, and offices, mostly with air conditioning systems, were locations in which air is not confined. This, added to frequent professional carpet cleaning, probably explains the lowest levels of fungal concentration found in these locations. The majority of the homes had the largest fungal concentration in the living-room (median: 2×10⁵ CFU/g) while some bedrooms (median: 7×10⁴ CFU/g) had the highest concentrations. It is suggested that, when fungi are suspected to be the origin of respiratory allergy or irritating symptoms, the mycoflora of the bedroom, principally, should be investigated first.     

Occurrence of toxigenic Aspergillus versicolor isolates and sterigmatocystin in carpet dust from damp indoor environments

Over the past decade, there has been growing concern regarding the role of toxigenic fungi in damp indoor environments; however, there is still a lack of field investigations on exposure to mycotoxins. The goal of our pilot study was to quantify the proportion of toxigenic Aspergillus versicolor isolates in native carpet dust from damp dwellings with mold problems and to determine whether sterigmatocystin can be detected in this matrix. Carpet dust samples (n = 11) contained from <2.5 x 10(1) to 3.6 x 10(5) (median, 3.1 x 10(4)) A. versicolor CFU/g of dust, and the median proportion of A. versicolor from total culturable fungi was 18%. Based on thin-layer chromatography detection of sterigmatocystin, 49 of 50 A. versicolor isolates (98%) were found to be toxigenic in vitro. By using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry, sterigmatocystin could be detected in low concentrations (2 to 4 ng/g of dust) in 2 of 11 native carpet dust samples. From this preliminary study, we conclude that most strains of A. versicolor isolated from carpet dust are able to produce sterigmatocystin in vitro and that sterigmatocystin may occasionally occur in carpet dust from damp indoor environments. Further research and systematic field investigation are needed to confirm our results and to provide an understanding of the health implications of mycotoxins in indoor environments.     

Effectiveness of vacuum cleaning and wet cleaning in reducing house-dust mites,fungi and mite allergen in a cotton carpet: A case study

In order to evaluate the effect of continuous, thorough vacuuming on house dust organisms and mite allergen, a cotton carpet was vacuumed every other day, six times in total. Seven weeks later, the carpet was cleaned by means of spray extraction. Samples were taken before and after this wet cleaning.In total 1150 g of dust was collected, containing approximately 174 000 arthropods (dead and alive) and 9000×10⁶ fungal spores. In the course of the experiment, the amount of dust collected at each vacuuming decreased. The number of extracted house dust mites did not decrease significantly, but that of the predator miteCheyletus did. The number of extracted fungal spores showed a significant decrease (from 142 to 16×10⁶ spores/m² per minute vacuuming), as did the extracted mite allergen per m². After 7 weeks the number of mite eggs and complete house dust mites had increased enormously. After cleaning by spray extraction another increase in the number of complete mites and mite eggs was found, while the amount of mite allergen was diminished.The population growth of the house dust mite between the 6th and the 7th vacuuming is probably due to the decrease of their most important predator,Cheyletus. After the wet cleaning a number of extra eggs hatched, probably due to the high humidity in the carpet. The procedures used in this study to combat house dust mites may have an adverse effect in the long run.     

Airborne dust,bacteria, actinomycetes and fungi at a flourmill

A study was carried out on suspended dust, bacterial and fungal aerosols in a four-storey flourmill building located in Giza, Egypt. Airborne microorganisms were quantitatively isolated using liquid impinger and gravimetric samplers during the period from March 2004 to February 2005. Suspended dust varied from 1.96 to 16.3 mg m⁻³ and 0.69 to 1.8 mg m⁻³ in the indoor and outdoor environments, respectively. Suspended dust was significantly greater (P < 0.05) at bran package, double roller, purifiers and flour storage units in comparison to the outdoor reference site. The dust levels exceed the occupational exposure limit (OEL) of 0.5 mg m⁻³ for flour dust. Airborne microbial counts were found at median values, between sampling locations, ranged from 0 to >10⁴ CFU m⁻³. Gram-negative bacteria were found in small numbers (0–10² CFU m⁻³). The highest concentration of actinomycetes (>10³ CFU m⁻³) was detected in the storage unit. Airborne fungal counts were found at the median values, between sampling locations, varied from 10³ to 10⁴ CFU m⁻³. The counts of airborne bacteria and fungi were significantly greater (P < 0.05) at the purifiers and double roller mill units in comparison to the outdoor reference site using the liquid impinger sampler. Microbial levels associated with bulk deposited dust averaged between 10⁵ and 10⁶ CFU g⁻¹. Alcaligenes (5.4%) Pseudomonas (3.87%) and Enterobacter (3.1%) were the predominant Gram-negative species while Bacillus (29.4%) and Micrococci (13.9%) were the major components of Gram-positive bacteria. Aspergillus and Penicillium were the predominant fungal types indoor whereas Cladosporium (35.2%) and Aspergillus species (22.2%) were the predominant fungal types outdoor. A number of allergenic and toxigenic bioaerosols were found in the flourmill workplace.     

Ochratoxin A in airborne dust and fungal conidia

Farm workers are often exposed to high concentrations of airborne organic dust and fungal conidia, especially when working with plant materials. The purpose of this investigation was to study the possibility of exposure to the mycotoxin ochratoxin A (OTA) through inhalation of organic dust and conidia. Dust and aerosol samples were collected from three local cowsheds. Aerosol samples for determination of total conidia and dust concentrations were collected by stationary sampling on polycarbonate filters. Total dust was analysed by gravimetry, and conidia were counted using scanning electron microscopy. A method was developed for extraction and determination of OTA in small samples of settled dust. OTA was extracted with a mixture of methanol, chloroform, HCI, and water, purified on immunoaffinity column, and analysed by ion-pair HPLC with fluorescence detection. Recovery of OTA from spiked dust samples (0.9–1.0 μg/kg) was 74% (quantitation limit 0.150 μg/kg). OTA was found in 6 out of 14 settled dust samples (0.2–70 μg/kg). The total concentration of airborne conidia ranged from < 1.1 × 10⁴ to 3.9 × 15⁵ per m³, and the airborne dust concentration ranged from 0.08 to 0.21 mg/m³. Conidia collected from cultures of Penicillium verrucosum and Aspergillus ochraceus contained 0.4–0.7 and 0.02–0.06 pg OTA per conidium, respectively. Testing of conidial extracts from these fungi in a Bacillus subtilis bioassay indicated the presence of toxic compounds in addition to OTA. The results show that airborne dust and fungal conidia can be sources of OTA. Peak exposures to airborne OTA may be significant, e.g., in agricultural environments. This revised version was published online in August 2006 with corrections to the Cover Date.     

A lack of influence of permethrin treatments of wool carpet on habitation by house dust mites

Wool carpet samples, untreated and treated with commercial levels of permethrin, were placed on a mite-inhabited carpet in a Melbourne house for 14 months. The levels of live house dust mites in the samples were found to be high at the end of this period and to exhibit no differences according to the presence of permethrin in the carpets.     

Measurement of airborne fungal spore dispersal from three types of flooring materials

Research was conducted in an experimental roomto measure the effect of human activity onairborne dispersal of settled fungal sporesfrom carpet and vinyl tile flooring. A seriesof experiments were conducted in whichcommercial loop pile carpet, residential cutpile carpet, or vinyl tile installed in theexperimental room were contaminated with Penicillium chrysogenum spores. The flooringmaterials were contaminated to two differentlevels (10⁶ and 10⁷ colony formingunits per square meter [c.f.u./m²] offlooring surface). Airborne culturable andtotal P. chrysogenum concentrations weremeasured using Andersen single-stage impactorsamplers and Burkard personal slide impactorsamplers, respectively. Bioaerosolconcentrations were measured at floor level, 1meter, and the adult breathing zone (1.5 meter)heights before and after human activityconsisting of walking in a prescribed patternfor 1 minute in the room. Airborne P.chrysogenum concentrations were greater withthe higher surface loading for all threeflooring materials. For all flooring materialsthere was no significant difference betweensampler locations, although the data from the1-meter location were the highest, followed bythe floor level and the breathing zonelocations, respectively. The data from theseexperiments indicate that while a very smallfraction of culturable P. chrysogenumspores present on flooring materials wereaerosolized by walking, relatively highairborne concentrations of spores maybere-entrained from contaminated materials. Theairborne P. chrysogenum concentrationswere significantly higher after walking on cutpile carpet than with the other two flooringmaterials at both contamination levels, withthe differences in concentration often 2orders of magnitude. No differences weremeasured in airborne culturable P.chrysogenum between vinyl flooring and looppile carpet at both contamination levels. Total spore data from the experiments with the10⁷ c.f.u./m² contamination levelindicated that walking on loop pile carpetproduced higher airborne spore concentrationsthan similarly contaminated vinyl tile althoughno significant difference was observed at the10⁶ c.f.u./m² level.     

Effect of entomopathogenic fungus <Emphasis Type="Italic">Metarhizium robertsii</Emphasis> on non-target organisms,water bugs (Heteroptera: Corixidae,Naucoridae, Notonectidae)

The influence of the fungus Metarhizium robertsii Bischoff, Rehner and Humber on the mortality of four water bug species, Cymatia coleoptrata (Fabricius), Sigara assimilis (Fieber), Ilyocoris cimicoides cimicoides (Linnaeus), and Notonecta reuteri Hungerford, and bloodsucking mosquito Anopheles messeae Falleroni, was investigated under various concentrations of conidia and different treatment types. We found that the mortality of adults of the water bug species was similar or higher than that of A. messeae, with C. coleoptrata and S. assimilis being more susceptible to M. robertsii than N. reuteri, I. c. cimicoides, and the mosquito A. messeae. Treatment with dry conidia at concentrations of 5 × 10⁴ and 5 × 10⁵ conidia/ml caused higher mortality of the water bug species than did treatment at the same concentrations with conidia in an aqueous suspension. In contrast, higher concentrations (5 × 10⁶ conidia/ml) led to higher mortality after treatment with the aqueous suspension, relative to treatment with dry conidia. Our studies showed that water bugs exhibited the classical development of a mycosis with hemocoel colonization, mummification, and conidia formation on cadavers directly on the surface of the water. Possible changes in invertebrate communities in aquatic ecosystems after treatment with Metarhizium are discussed.     

Bioavailability and speciation of arsenic in urban street dusts from Baoding city,China

Twenty-one street dust samples were collected from the urban sites in Baoding, China, to investigate the species and bioavailabilty of arsenic in them. The ecological risk and bioavailability of arsenic were evaluated using three models including Bioavailability Factor (BF), Contamination Factors (C_f)and Geoaccumulation Index model (I_geo). The species of arsenic in the dust samples were analyzed using an optimized BCR sequential extraction method. The total concentrations of As in the street dust samples ranged from 13.16 mg kg^?1 to 67.26 mg kg^?1. Geoaccumulation index (I_geo) of As ranged from 0.28 to 1.99. The speciation analysis indicated that As in the street dust samples were mainly in the residual fraction (F4), and the proportion ranged from 84.35% to 87.07%. Moreover, the ranges of the BF and C_f were 0.650–0.129 and 0.119–0.186, respectively. The results indicated that arsenic contained in the street dust samples was with low bioavailability.     

Scanning electron microscopy of Penicillium conidia

The morphology of conidia in 211 species and 12 varieties belonging to the genus Penicillium Link ex Gray have been studied and compared.According to surface ornamentation, conidia have been classified into six groups: A, smooth-walled (7% of the species); B, delicately roughened (13%); C, warty (28%); D, echinate (10%); E, striate with low irregular ridges (36%); and F, striate with scarce high ridges or bars (6%). Whereas the first two groups are closely related in both shape and average size, a gradual reduction was observed in size and in the length/width (l/w) ratio in the remaining groups. Echinate conidia were globose, having the largest average size. Only four species produced conidia not surpassing 2 m in diameter. Maximum length observed was 8 m, and most elongated conidia had a l/w ratio of 3.5. Forty per cent of the species studied had globose conidia.Conidia of the monoverticillate species were generally smaller, more globose and frequently with ridges. In the Asymmetrica, the conidia were generally larger, and showed ridges in comparatively few species. Conidia of the Symmetrica, which were frequently striate with ridges, presented the most elongated forms. The largest average size was found in the conidia of the Polyverticillata which were generally warty. Finally, we have considered the variations in surface ornamentation of conidia during the evolution of the genus Penicillium and drawn attention to their possible relationship with certain habitats and ways of conidial dispersion.     

Dustborne fungi in large office buildings

Fungi are ubiquitous in our daily environments. However, their effects on office workers' health are of great interest to many environmental health researchers. Dust has been considered an important reservoir of indoor fungi from which aerosolization and exposure could occur. We have examined the characteristics of dustborne fungal populations recovered from floors and chairs in office buildings. We investigated twenty-one offices in four office buildings in Boston, MA over a year beginning May 1997. We conducted intensive environmental sampling every six weeks to measure culturable dustborne fungi from floors and chairs, surface dust levels and water activity in carpeting. Carbon dioxide, temperature, and relative humidity were monitored continuously. Concentrations of total dustborne fungi recovered from floors were positively related to carbon dioxide ( = 0.00064; p-value = 0.0002) and temperatures between 20 and 22.5 °C (p-value = 0.0026). Also, total fungal concentrations in floors gradually increased over the year (p-value = 0.0028). Total fungi recovered from chairs varied significantly by season (p-value < 0.0001),highest in September and lowest in March, and were positively correlated with dust loads in floors ( = 0.25; p-value < 0.0001). We used principal component analysis (PCA) to reduce various observed fungal species to fewer factors. Six groups(PCA factors) were obtained for dustborne fungi recovered from both floors and chairs. The models of the first PCA factors for both floors and chairs were similar to those for total fungal concentrations. The results of this study provide essential information to further evaluate the effects of dustborne fungi on office workers' health.This revised version was published online in October 2005 with corrections to the Cover Date.     

Attachment of Blastomyces dermatitidis conidia to murine bronchoalveolar macrophages: Characterization of binding the elicitation of respiratory burst

We studied the mechanisms of adherence of Blastomyces dermatitidis conidia to murine bronchoalveolar macrophages and the ability of the conidia to elicit an increase in macrophage O _inf2 ^sup- production, using an avirulent fungal strain. The number of cell associated conidia was counted by visual inspection of 2 hour macrophage monolayers incubated with conidia and O _inf2 ^sup- was measured by reduction of ferricytochrome c. Adherence of conidia to bronchoalveolar macrophages was time dependent and reached a plateau after 30 min (36±5%, 51±22%, and 36±17% macrophages with adherent conidia after 15, 30, and 60 min, respectively). Both Ca⁺² and Mg⁺² were required. The carbohydrates mannose, mannan, fucose, alpha-methylmannoside, beta-glucan, galactose, N-acetylglucosamine and chitotriose (100–1000 g/ml) did not inhibit adherence of conidia to macrophages. Trypsin treatment of macrophages or conidia did not affect binding. Conidia did not stimulate bronchoalveolar macrophage production of O _inf2 ^sup- above baseline concentrations (2.0±0.9 vs 0.8±0.5 nmol O _inf2 ^sup- , p>0.05). We conclude that murine bronchoalveolar macrophage-B. dermatitidis conidia interactions occur primarily by a non-lectin-like attachment and do not result in the production of macrophage derived O _inf2 ^sup- .     

Inhibition of some mycotoxigenic fungi by iturin A,a peptidolipid produced by Bacillus subtilis

Bacillus subtilis produces peptidolipid compounds of the iturin group that have been shown to have antifungal properties, but not all fungal species are sensitive to these compounds. In this study, the activity of iturin A, produced by B. subtilis strain B-3, was tested. Paper disks impregnated with various concentrations of iturin A were placed on agar plates seeded with conidia of toxigenic species of Fusarium, Gerlacia, Penicillium or Aspergillus. Most isolates were inhibited at iturin A concentrations as low as 4 g/disk. Penicillium italicum, P. vindicatum, A. ochraceus and A. versicolor were most strongly inhibited by the iturin whereas P. citrinum and A. parasiticus were least sensitive to iturin A.Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the US Department of Agriculture and does not imply approval to the exclusion of other products that may also be suitable.     

Mould biodiversity in homes II. Analysis of mattress dust

Fungal species from 130 samples of mattress dust have been identified. The number of moulds varies from one mattress to another with extreme values ranging from less than 1000 to more than 70 000 000 CFU/g of dust (median value of 30 000 CFU/g).Eurotium repens is undoubtedly the most frequent species detected in mattress dust.Aureobasidium pullulans, Alternaria alternata, Penicillium chrysogenum, Aspergillus penicilloides andAspergillus restrictus, which were frequently isolated and/or revealed important concentrations in dust samples, are the other main occurring species. Fungal flora in mattresses are more numerous in winter, the peak period being December-January. The most important contaminations by xerophilic species occurred during that peak period, whereas non-osmophilous species were constantly present during the year. The qualitative conditions of the habitat have an influence on the abundance of moulds in mattresses. Mattresses from damp houses could be a source forAlternaria throughout the year. No significant difference has been detected between the contamination level of men and women's mattresses (P=0.651). Children's mattresses are less contaminated than adults (P=0.025). The level of contamination could be affected by perspiration.     

Pupation site selection of cat fleas (Siphonaptera: Pulicidae) in various carpet types and its influence on insecticide efficacy

Pupation sites of cat flea, Ctenocephalides felis (Bouché), larvae were determined in three styles of nylon and one style of wool carpet. Nylon saxony carpet had 59.3% of pupae at the top of the pile and 40% at the base of the pile. In nylon contract carpet, 55.2% of pupae were found at the top, 42.6% in the middle, and only 2.2% at the base of the pile. Nylon loop carpet contained 59.2% of pupae at the base, 25.5% in the middle, and 15.3% in the top of the pile. Wool loop carpet had 92.4% at the base and 3.8% both in the middle and top of the pile. Bioassays comparing the control of pupae manually placed at the base of carpets to that in carpets with natural pupation showed that control of pupae in the latter was 39-68% higher. Pupal control after natural pupation was greatest in nylon saxony and nylon contract carpets and lowest in nylon loop and wool loop carpets. Additional studies demonstrated that vacuuming provided the same level of pupal control on nylon saxony carpet as a spray application of permethrin to the carpet surface. Therefore, pupae that survived chemical and mechanical control treatments in nylon saxony carpet probably pupated away from the surface of the pile. Application of permethrin to the base of nylon saxony carpet did not significantly increase control. Future bioassays with cat flea pupae in carpet should be performed after natural pupation and consider carpet make and style.     

Adhesion,spreading, and proliferation of cells on protein carpets: Effects of stability of a carpet

Summary In the present report we have investigated the role that the physical properties of substrata play in modulating the effects which components of extracellular matrix (ECM) exert on adhesion, spreading, and growth of retinal pigmented epithelial cells. By simple modifications of conditions for protein adsorption on glass we obtained a set of substrata all coated with proteins of ECM (protein carpets) but with different physical properties. Using these protein carpets we have shown that their stability (desorption rate) in tissue culture conditions varies according to the technique with which they were prepared. Both semiremovable and immobilized carpets are stable, whereas removable protein carpets desorb readily. Therefore, the protein concentration or composition or both may change with time in tissue culture depending on the technique used to prepare the carpet. In addition, efficacy of cell attachment to given protein may vary depending on whether a technique used to prepare the protein carpet involves denaturation of the protein. Adherent cells quickly remove (clear) weakly adsorbed protein carpets and it seems that the carpet removal is a mechanical process. During the carpet removal cells are rounded, which indicates that a spread cell phenotype normally associated with stress fibers and focal contacts occurs when the substratum is rigid enough to sustain cell traction. In addition, substrata lacking the rigidity to support the spread phenotype do not support cell proliferation either.     

Myrothecium: A new indoor contaminant?

In the natural environmentMyrothecium species occur as soil or leaf surface saprobes or as weak plant pathogens. In addition, some species ofMyrothecium are known to produce trichothecene mycotoxins. During a previous aerobiological investigation at two Las Vegas elementary schools,Myrothecium conidia were found to be the second most abundant spore type identified indoors from Burkard personal spore trap samples. The present study was undertaken to re-examine the schools to locate the source ofMyrothecium spores and to examine the ability ofMyrothecium to grow on indoor substrates. There were no obvious signs of contamination in the classrooms; however,Myrothecium spores occurred on about 30% of the Burkard samples. Two colonies ofMyrothecium were identified from subcultures of the Andersen samples, and three colonies were identified from carpet dust samples. Culture studies showed that a strain ofMyrothecium cinctum was able to grow on various culture media as well as on various indoor substrates including paper, cardboard, wallpaper, ceiling tiles, dry wall, carpets and cotton rug. Although there was no attempt to estimate any human health risks, these investigations are believed to be the first to document abundantMyrothecium spores from indoor air samples.     

Survival and proliferation of alginate encapsulated Trichoderma spp. in Egyptian soil in comparison with allyl alcohol soil fumigation

Conidia of Trichoderma harzianum and T. pseudokoningii (Rifai) were formulated to make alginate pellets withor without 10% cellulose as a food-base material. The formulations were compared for their ability for survival and proliferation of Trichoderma spp. in clay-loamy soil (50% moisture content)with allyl alcohol fumigation (0.05, 0.1 and 0.2 ml/1000 ml space). Trichoderma medium E (TME) containing 100 g/ml pentachloronitrobenzene (PCNB) was valuable for isolation and counting of Trichoderma spp. from the tested soil than the Glucose-Czapek's agar medium containing 1:15000 Rose-bengal. The promotive effect of Trichoderma by different doses of allyl alcohol fumigationstill enhanced after two-month incubation period. Conidia entrapped in alginate with or without cellulose and introduced into the soil survived better than conidia added directly to the same soil after three months incubation period. Sterile soil provided a more favorable environment for the proliferation and survival of immobilized conidia than the non-sterile soil, and the addition of 10% cellulose increased the survival of the entrapped conidia more than those prepared without cellulose. Soil fumigation inhibited the occurrence of other fungal species; however, inoculation of the soil with alginate immobilized conidia or conidial suspension had such inhibitory effect but in a less extent.This revised version was published online in October 2005 with corrections to the Cover Date.