Increased Levels of Markers of Microbial Exposure in Homes with Indoor Storage of Organic Household Waste

As part of environmental management policies in Europe, separate collection of organic household waste and nonorganic household waste has become increasingly common. As waste is often stored indoors, this policy might increase microbial exposure in the home environment. In this study we evaluated the association between indoor storage of organic waste and levels of microbial agents in house dust. The levels of bacterial endotoxins, mold β(1→3)-glucans, and fungal extracullar polysaccharides (EPS) of Aspergillus and Penicillium species were determined in house dust extracts as markers of microbial exposure. House dust samples were collected in 99 homes in The Netherlands selected on the basis of whether separated organic waste was present in the house. In homes in which separated organic waste was stored indoors for 1 week or more the levels of endotoxin, EPS, and glucan were 3.2-, 7.6-, and 4.6-fold higher, respectively (all P < 0.05), on both living room and kitchen floors than the levels in homes in which only nonorganic residual waste was stored indoors. Increased levels of endotoxin and EPS were observed, 2.6- and 2.1-fold (P < 0.1), respectively, when separated organic waste was stored indoors for 1 week or less, whereas storage of nonseparated waste indoors had no effect on microbial agent levels (P > 0.2). The presence of textile floor covering was another major determinant of microbial levels (P < 0.05). Our results indicate that increased microbial contaminant levels in homes are associated with indoor storage of separated organic waste. These increased levels might increase the risk of bioaerosol-related respiratory symptoms in susceptible people.     

Diversity and Significance of Mold Species in Norwegian Drinking Water

In order to determine the occurrence, distribution, and significance of mold species in groundwater- and surface water-derived drinking water in Norway, molds isolated from 273 water samples were identified. Samples of raw water, treated water, and water from private homes and hospital installations were analyzed by incubation of 100-ml membrane-filtered samples on dichloran-18% glycerol agar. The total count (number of CFU per 100 ml) of fungal species and the species diversity within each sample were determined. The identification of mold species was based on morphological and molecular methods. In total, 94 mold species belonging to 30 genera were identified. The mycobiota was dominated by species of Penicillium, Trichoderma, and Aspergillus, with some of them occurring throughout the drinking water system. Several of the same species as isolated from water may have the potential to cause allergic reactions or disease in humans. Other species are common contaminants of food and beverages, and some may cause unwanted changes in the taste or smell of water. The present results indicate that the mycobiota of water should be considered when the microbiological safety and quality of drinking water are assessed. In fact, molds in drinking water should possibly be included in the Norwegian water supply and drinking water regulations.     

Pilot study of mold populations inside and outside of Puerto Rican residences

Puerto Rico has the highest asthma prevalence in the US. In the states, mold exposures have been linked to the development and exacerbation of asthma. For a pilot study of mold populations in Puerto Rico, dust and air samples were collected in January 2013 inside and outside of residences (n = 10) in the north eastern quadrant of Puerto Rico. Inside and outside (protected from rain) settled dust was collected by wiping above floor or ground-level surfaces with a Swiffer? duster cloth. Air samples were collected in five of the residences inside and five matching outside onto Malt Extract Agar and also onto G25N Agar. The dust (5 mg) was analyzed using mold-specific quantitative PCR and mold contamination estimated using the Environmental Relative Moldiness Index (ERMI) metric. Air sampled (150 l) plates were incubated and resulting mold colonies enumerated. The ERMI values were significantly (p = 0.03) greater inside the homes than outside, 20.37 versus 12.55, respectively. Aspergillus penicillioides, A. sydowii, Chaetomium globosum, the Eurotium group and Penicillium crustosum and A. ustus were found in significantly greater numbers inside than outside Puerto Rican residences based on the Wilcoxon rank sum test. However, after correction for multiple comparisons using the Holms–Bonferroni test, only the Eurotium group remained significantly greater. Air samples suggested higher culturable concentrations of molds outside but the differences were not significantly different. Although this is a small study, it suggests that some people in Puerto Rico are exposed to significantly higher mold concentrations indoors than outdoors.     

Evaluation of bio-aerosols at an animal feed manufacturing industry: A case study

Both total and biological particles (totalculturable bacteria, Gram negative bacteria,mold and actinomycetes) were measured at ananimal feed manufacturing industry. Suspendedparticle concentration ranged from 1.72 to2.3 mg m^–3 with a mean value of1.97 mg m^–3. Airborne microorganisms weredetected in lower concentrations than thoseassociated with suspended dust andfeed-materials. Bacterial concentrations weretwo to three times higher than concentrationsof mold and actinomycetes. Bacterialconcentrations averaged4.86 × 10³ cfu m^–3; 2.6 × 10⁴cfu m^–3 and 3.96 × 10⁷ cfu g^–1 inair, associated with suspended dust andfeed-materials, respectively, whereas moldconcentrations averaged 7.33 × 10²cfu m^–3; 1.97 × 10³ cfu m^–3 and7 × 10⁵ cfu g^–1 of the correspondingenvironments, respectively. Enterobacterspp and Klebsiella spp were the mostabundant Gram negative bacteria, whereas Bacillus species. were the most dominant Grampositive bacteria. Aspergllius niger,other Aspergillus species and Penicillium were the dominant mold isolates.Acremonium was only detected in feedmaterials, whereas Aspergillus fumigatuswas only detected in air. The animal feedindustry environment has a significantbio-contamination and many of microorganismsimplicated in respiratory problems weredetected in this environment.     

Mycoflora isolation and molecular characterization of Aspergillus and Fusarium species in Tunisian cereals

Wheat, barley and maize are the mainly consumed cereals in Tunisia. This study aimed to determine the mycoflora of these cereals with special focus on the mycotoxigenic Aspergillus and Fusarium species. Freshly harvested samples and other stored samples of each type of cereal (31 and 34 samples, respectively) were collected in Tunisia and cultured for fungal isolation and identification. Identification of fungal genera was based on morphological features. Aspergillus and Fusarium species were identified by species specific PCR assays complemented with DNA sequencing. Alternaria (70.83%), Eurotium (62.50%), Aspergillus (54.17%) and Penicillium (41.67%) were the most frequent fungi isolated from wheat. Penicillium (75%), Aspergillus (70%), Eurotium (65%) and Alternaria (65%) were the most frequently recovered genera from barley. The predominant genera in maize were Aspergillus (76.19%), Eurotium (42.86%), and Penicillium (38.09%). Aspergilllus, Penicillium, Fusarium and Alternaria were detected in both stored and freshly harvested grain samples. The frequencies of contamination with Aspergillus, Fusarium and Alternaria were higher in freshly harvested samples, whereas Penicillium species were more frequent in stored samples. The predominant Aspergillus species detected were A. flavus and A. niger. The Fusarium species detected were F. equiseti, F. verticillioides, F. nygamai, and F. oxysporum. This study suggested the potential risk for Aflatoxins and, to a lesser extent, for Ochratoxin A in Tunisian cereals. This is the first survey about mycoflora associated with wheat, barley and maize in Tunisia.     

Seasonal Variations of Indoor Microbial Exposures and Their Relation to Temperature,Relative Humidity,and Air Exchange Rate

Indoor microbial exposure has been related to adverse pulmonary health effects. Exposure assessment is not standardized, and various factors may affect the measured exposure. The aim of this study was to investigate the seasonal variation of selected microbial exposures and their associations with temperature, relative humidity, and air exchange rates in Danish homes. Airborne inhalable dust was sampled in five Danish homes throughout the four seasons of 1 year (indoors, n = 127; outdoors, n = 37). Measurements included culturable fungi and bacteria, endotoxin, N-acetyl-beta-d-glucosaminidase, total inflammatory potential, particles (0.75 to 15 μm), temperature, relative humidity, and air exchange rates. Significant seasonal variation was found for all indoor microbial exposures, excluding endotoxin. Indoor fungi peaked in summer (median, 235 CFU/m³) and were lowest in winter (median, 26 CFU/m³). Indoor bacteria peaked in spring (median, 2,165 CFU/m³) and were lowest in summer (median, 240 CFU/m³). Concentrations of fungi were predominately higher outdoors than indoors, whereas bacteria, endotoxin, and inhalable dust concentrations were highest indoors. Bacteria and endotoxin correlated with the mass of inhalable dust and number of particles. Temperature and air exchange rates were positively associated with fungi and N-acetyl-beta-d-glucosaminidase and negatively with bacteria and the total inflammatory potential. Although temperature, relative humidity, and air exchange rates were significantly associated with several indoor microbial exposures, they could not fully explain the observed seasonal variations when tested in a mixed statistical model. In conclusion, the season significantly affects indoor microbial exposures, which are influenced by temperature, relative humidity, and air exchange rates.     

Winged bean (Psophocarpus tetragonolobus (L.) DC) as a substrate for growth and aflatoxin production by aflatoxigenic strains of Aspergillus spp.

The mold flora of seeds of twelve varieties of winged beans were determined both before and after surface disinfections. When seeds were surface disinfected, molds were detected in 73% of the seeds whereas 81% of the seed that was not disinfected produced molds. Aspergillus spp. were most frequently present while Penicillium spp. occurred in seed of 4 varieties and in less than 4% of the seed. Twelve isolates of A. flavus and A. parasiticus were examined for their ability to produce aflatoxins. Whether aflatoxins were produced and the amount of each varied according to the origin of the isolate and the species of Aspergillus. For example all A. flavus isolates produced at least 2 aflatoxins whereas 4 of the A. parasiticus isolates were nontoxigenic. When ground seeds of winged beans were inoculated with an aflatoxigenic strain of A. parasiticus the level of aflatoxins that occurred varied with the variety. All of the varieties supported greater aflatoxin production than peanuts and 6 of the 12 winged bean varieties gave higher levels of aflatoxins than rice.     

Durability of rubberwood (Hevea brasiliensis) treated with peppermint oil,eucalyptus oil,and their main components

Anti-fungal activities of two essential oils (peppermint oil and eucalyptus oil) and their main components (menthol and eucalyptol, respectively) against molds (Aspergillus niger, Penicillium chrysogenum, and Penicillium sp.) and a white-rot decay fungus (Trametes versicolor) identified from rubberwood surfaces were investigated. The broth dilution method and the agar diffusion technique were employed to determine the minimal inhibitory concentration (MIC) and the minimal fungicidal concentration (MFC) using the concentration of substances between 100 and 800 μl ml^?1. Inhibitory effects of essential oils and their main components at the MICs against mold growth, fungal decay, and termite attack on rubberwood were further examined by means of the dip treatment method. It was found that MFC values against molds for all treatments examined were about 50–100 μl ml^?1 higher than MIC values. Peppermint oil and menthol exhibited high fungistatic and fungicidal activities, with MICs of 300 μl ml^?1 and 350 μl ml^?1, respectively, against the test molds and the decay fungus. Eucalyptus oil and eucalyptol were also effective against these microbes but at higher concentrations of 600 μl ml^?1 and 500 μl ml^?1, respectively. Only peppermint oil at the MIC was capable of providing a complete protection from mold growth on rubberwood for up to 12 weeks at storage conditions of 25 °C and 100% RH. Both peppermint oil and eucalyptus oil at the MICs showed moderate resistance to fungal decay and high resistance to termite attack.     

Airborne fungi associated with ornamental plant propagation in greenhouses

The objective was to determine potential exposure to airborne fungi in greenhouses and to characterize the temporal patterns of airborne fungi in relation to environmental conditions. We analyzed air samples collected in two greenhouses. Results showed that the top 5 fungi in greenhouse 1 were Trichoderma, hyphal fragments, Aspergillus/Penicillium-like, Cladosporium, and Botrytis in a descending order. Those in greenhouse 2 were Aspergillus/Penicillium-like, Cladosporium, Botrytis, yeast-like, and hyphal fragments. Maximum concentrations of Trichoderma and total spores in greenhouse 1 were 36,426 and 49,729 spores/m³, respectively. Maximum concentrations of Aspergillus/Penicillium and total spores in greenhouse 2 were 46,961 and 71,037 spores/m³, respectively. Airborne fungal populations fluctuated dramatically within 2 h during work hours, tenfold for Aspergillus/Penicillium, 66-fold for Trichoderma, and sevenfold for total spores. QPCR detected Trichoderma harzianum ranging from 7 to 3,500 conidia E/m³. Aspergillus/Penicillium and Botrytis showed diurnal patterns, but not Trichoderma. Aspergillus/Penicillium and Cladosporium were positively correlated with temperature, relative humidity, dew point, heat index, and light and negatively with air movement and air pressure. Botrytis and Trichoderma were not correlated with the environmental factors. Greenhouse workers were potentially exposed up to 71,037 spores/m³ of airborne fungi.     

Caracterización de la micobiota aérea en dos depósitos del Archivo Nacional de la República de Cuba

BackgroundThe high relative humidity and temperatures in tropical countries create favorable conditions for development of fungi that are not only a risk to human health but they can also colonize documentary support.AimsTo study the concentration of the airborne fungi in two repositories of the National Archives of the Republic of Cuba, the mycobiota deposited on different photographic supports and maps preserved in these repositories, and to determine the taxonomic characterization of the fungi isolated.MethodsThe air sampling was performed using a sedimentation method, and the supports (6 pictures and 7 maps) were analyzed using moistened sterile swabs.ResultsThe Cladosporium genus was predominant, followed by Aspergillus and Penicillium genera. Filamentous fungi were isolated in all the photos and maps, and yeasts were only isolated from a photographic supports and a map. We identified several species of Aspergillus and Penicillium genera, but Aspergillus niger and Aspergillus flavus predominated. Candida and Rhodotorula were the yeast genera isolated.ConclusionsThe fungal concentration of the air demonstrated that the environments were not contaminated. From the 26 species of filamentous fungi isolated only 5 were detected in the indoor air of the repositories and on one or more of the document supports analyzed (representing a 19.3%). This shows that there is a low ecological relationship between the fungi detected in the indoor air and those that were isolated from the supports studied.     

Mould in buildings: the air spora of domestic dwellings

Filamentous fungi in 37 genera were isolated during winter from the internal air of 62 domestic dwellings in England and Scotland. In the air spora the number of viable fungal colony-forming units (cfu), including yeasts, ranged from < 12 to 449 800 cfu m⁻³. There was considerable variation between dwellings and between weekly samples taken within the same dwelling, but in approximately one-third of all air samples there were < 200 cfu m⁻³. The filamentous fungi isolated most frequently from the air in 47 Scottish dwellings were Penicillium spp. (96% of dwellings), Cladosporium spp. (89%), Aspergillus spp. (75%, mainly A. versicolor), Ulocladium spp. (62%), Geomyces pannorum (57%) and Sistotrema brinkmannii (51%). Yeasts were isolated in 94% of these dwellings and overall comprised 13% of cfu on Andersen sampler plates. Penicillium spp., Cladosporium spp. and S. brinkmannii together accounted for > 70% of the propagules. High numbers of spores in the internal air were associated with surface mould growth and constructional work. In addition, disturbance of surface growth and vacuum cleaning of carpets caused large temporary increases in the atmospheric spore count.     

Fungal Fragments as Indoor Air Biocontaminants

The aerosolization process of fungal propagules of three species (Aspergillus versicolor, Penicillium melinii, and Cladosporium cladosporioides) was studied by using a newly designed and constructed aerosolization chamber. We discovered that fungal fragments are aerosolized simultaneously with spores from contaminated agar and ceiling tile surfaces. Concentration measurements with an optical particle counter showed that the fragments are released in higher numbers (up to 320 times) than the spores. The release of fungal propagules varied depending on the fungal species, the air velocity above the contaminated surface, and the texture and vibration of the contaminated material. In contrast to spores, the release of fragments from smooth surfaces was not affected by air velocity, indicating a different release mechanism. Correlation analysis showed that the number of released fragments cannot be predicted on the basis of the number of spores. Enzyme-linked immunosorbent assays with monoclonal antibodies produced against Aspergillus and Penicillium fungal species showed that fragments and spores share common antigens, which not only confirmed the fungal origin of the fragments but also established their potential biological relevance. The considerable immunological reactivity, the high number, and the small particle size of the fungal fragments may contribute to human health effects that have been detected in buildings with mold problems but had no scientific explanation until now. This study suggests that future fungal spore investigations in buildings with mold problems should include the quantitation of fungal fragments.     

Fungi Isolated from Flue-cured Tobacco at Time of Sale and After Storage

The fungi isolated from 100 samples of flue-cured tobacco from 12 markets in 2 tobacco belts comprised 11 genera, including 10 species of Aspergillus. The mean percentage per sample isolated from 62 samples of tobacco from Middle Belt markets was Alternaria, 40.6%; Aspergillus niger, 47.8%; Aspergillus repens, 38.0%; and Penicillium, 25.8%. The mean percentage per sample isolated from 38 samples of tobacco from Old Belt markets was Alternaria, 74.0%; Penicillium, 52.5%; Aspergillus repens, 38.0%; and Aspergillus ruber, 36.2%. Damaged (74 samples) and nondamaged (26 samples) stored tobacco yielded species of six genera of fungi, including eight species of Aspergillus. Species of Aspergillus and Penicillium were commonly isolated from both damaged and nondamaged tobacco, whereas species of Alternaria, Cladosporium, Fusarium, and Rhizopus were isoalted more frequently from nondamaged tobacco. The fungi that occurred in the highest population in damaged tobacco were Aspergillus repens, A. niger, A. ruber, and Penicillium species.     

Metagenomic Insights into the Bioaerosols in the Indoor and Outdoor Environments of Childcare Facilities

Airborne microorganisms have significant effects on human health, and children are more vulnerable to pathogens and allergens than adults. However, little is known about the microbial communities in the air of childcare facilities. Here, we analyzed the bacterial and fungal communities in 50 air samples collected from five daycare centers and five elementary schools located in Seoul, Korea using culture-independent high-throughput pyrosequencing. The microbial communities contained a wide variety of taxa not previously identified in child daycare centers and schools. Moreover, the dominant species differed from those reported in previous studies using culture-dependent methods. The well-known fungi detected in previous culture-based studies (Alternaria, Aspergillus, Penicillium, and Cladosporium) represented less than 12% of the total sequence reads. The composition of the fungal and bacterial communities in the indoor air differed greatly with regard to the source of the microorganisms. The bacterial community in the indoor air appeared to contain diverse bacteria associated with both humans and the outside environment. In contrast, the fungal community was largely derived from the surrounding outdoor environment and not from human activity. The profile of the microorganisms in bioaerosols identified in this study provides the fundamental knowledge needed to develop public health policies regarding the monitoring and management of indoor air quality.     

Presence ofAspergillus Mich. ex Fr. in several different biotopes in Córdoba (Spain)

The medical, veterinary and saprophytic importance ofAspergillus has been acknowledged by many authors. This paper reports on the occurrence ofAspergillus species in the air and dust of Córdoba (Spain). Four studied were done: first, on the air of three rooms and the outside area of 14 homes over 2 years; second, outdoor air in the west of the central district, with three samples daily over 1 year; third, the air inside and outside four of the largest silos and commercial grain warehouses in the province, over 1 year: and fourth, the dust of 12 primary schools in Córdoba city, for two consecutive school years. For air-sampling, volumetric and gravimetric methods were used, both with a culture medium and with 20-min exposure. For dust-sampling, a customised vacuum cleaner was used; samples collected were inoculated onto Petri dishes containing culture medium. A total of 38Aspergillus species were identified (we also detected four ascosporic states). A comparison of the results obtained at each of the sampling sites was made and an analysis of variance revealed quantitatively significant differences between sampling times.     

Variability in airborne bacterial and fungal population in the tertiary health care centre

An investigation of the air quality and the quantity of airborne microbes was conducted in a private and a government tertiary health care centre of Davanagere in the month of November 2011 to assess the level of air borne pathogens. Using a Merck Microbial Air Sampler MAS-100NT, samples were collected in the morning and in the evening from the different environs of the private and government tertiary health care centre. The media used for the study of fungi was sabouraud dextrose agar. Aspergillus spp, Curvularia spp, Alternaria spp, Penicillium spp, Rhizopus spp, Nigrospora spp, and Fusarium spp were found in either of the tertiary health care centre. Aspergillus spp was dominant in the Government tertiary health care centre, and Alternaria spp and Curvularia spp were dominant in the private tertiary health care centre. For the bacteria, quantitative enumeration was done using soyabean casein digest agar and selective media like Escherichia coli and coliform agar and urinary tract infection agar were used in qualitative enumeration. Selected pathogens like E. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enteritidis, Staphylococcus aureus, Proteus mirabilis, and Entirococcus faecalis were found in either of the tertiary health care centre. Maximum number of fungi and bacteria were isolated from emergency ward and general ward of government and private health care centre. There was also considerable difference in the morning and in the evening.     

Phyllosphere microflora of some Egyptian plants

The fungal and bacterial flora of the leaf surfaces of five plants growing in Egypt were studied. The fungal flora showed seasonal fluctuations with at least one peak. Twenty three genera with fifty three species were found,Aspergillus andPenicillium being most common. Other fungi showed variable percentages of the total count. Nitrogen-fixing bacteria were not isolated from the phyllosphere of the five plant species. Micrococci were most predominant among the epiphytic bacteria. Spore-forming bacteria and actinomycetes were less frequent on the leaf surfaces of the associated plants.     

Isolation and Characterization of Microorganisms Associated with the Traditional Sorghum Fermentation for Production of Sudanese Kisra

Sorghum flour obtained from Sudan was mixed with water in a 1:2 (wt/vol) ratio and fermented at 30°C for 24 h. The bacterial populations increased with fermentation time and reached a plateau at approximately 18 h. At the end of 24 h, sorghum batter pH had dropped from 5.95 to 3.95 and the batter had a lactic acid content of 0.80%. The microbial population during the 24 h of fermentation consisted of bacteria (Pediococcus pentosaceus, Lactobacillus confusus, Lactobacillus brevis, Lactobacillus sp., Erwinia ananas, Klebsiella pneumoniae, and Enterobacter cloacae), yeasts (Candida intermedia and Debaryomyces hansenii), and molds (Aspergillus sp., Penicillium sp., Fusarium sp., and Rhizopus sp.). P. pentosaceus was the dominant microorganism at the end of the 24-h fermentation. When three consecutive fermentations using an inoculum from the previous fermentation were carried out, the bacterial population increase plateaued at 9 h. The microbial populations in these fermentations were dominated by P. pentosaceus.     

Identifying airborne fungi in Seoul,Korea using metagenomics

Fungal spores are widespread and common in the atmosphere. In this study, we use a metagenomic approach to study the fungal diversity in six total air samples collected from April to May 2012 in Seoul, Korea. This springtime period is important in Korea because of the peak in fungal spore concentration and Asian dust storms, although the year of this study (2012) was unique in that were no major Asian dust events. Clustering sequences for operational taxonomic unit (OTU) identification recovered 1,266 unique OTUs in the combined dataset, with between 223?96 OTUs present in individual samples. OTUs from three fungal phyla were identified. For Ascomycota, Davidiella (anamorph: Cladosporium) was the most common genus in all samples, often accounting for more than 50% of all sequences in a sample. Other common Ascomycota genera identified were Alternaria, Didymella, Khuskia, Geosmitha, Penicillium, and Aspergillus. While several Basidiomycota genera were observed, Chytridiomycota OTUs were only present in one sample. Consistency was observed within sampling days, but there was a large shift in species composition from Ascomycota dominant to Basidiomycota dominant in the middle of the sampling period. This marked change may have been caused by meteorological events. A potential set of 40 allergyinducing genera were identified, accounting for a large proportion of the diversity present (22.5?7.2%). Our study identifies high fungal diversity and potentially high levels of fungal allergens in springtime air of Korea, and provides a good baseline for future comparisons with Asian dust storms.