Presence and variability of culturable bioaerosols in three multi-family apartment buildings with different ventilation systems in the Northeastern US.

Bioaerosol concentrations in residential buildings located in the Northeastern US have not been widely studied. Here, in 2011-2015, we studied the presence and seasonal variability of culturable fungi and bacteria in three multi-family apartment buildings and correlated the bioaerosol concentrations with building ventilation system types and environmental parameters. A total of 409 indoor and 86 outdoor samples were taken. Eighty-five percent of investigated apartments had indoor-outdoor (I/O) ratios of culturable fungi below 1, suggesting minimal indoor sources of fungi. In contrast, 56% of the apartments had I/O ratios for culturable bacteria above 1, indicating the prominence of indoor sources of bacteria. Culturable fungi I/O ratios in apartments serviced by central heating, ventilation, and air-conditioning (HVAC) system were lower than those in apartments with window AC. The type of ventilation system did not have a significant effect on the presence of indoor culturable bacteria. A significant positive association was determined between indoor dew point (DP) levels and indoor culturable fungi (P < .001) and bacteria (P < .001), regardless of ventilation type. Also, residents in apartments with central HVAC did not experience extreme DP values. We conclude that building ventilation systems, seasonality, and indoor sources are major factors affecting indoor bioaerosol levels in residential buildings.

Uncovering fungal community composition in natural habitat of Ophiocordyceps sinensis using high-throughput sequencing and culture-dependent approaches.

BACKGROUND: The fungal communities inhabiting natural Ophiocordyceps sinensis play critical ecological roles in alpine meadow ecosystem, contribute to infect host insect, influence the occurrence of O. sinensis, and are repertoire of potential novel metabolites discovery. However, a comprehensive understanding of fungal communities of O. sinensis remain elusive. Therefore, the present study aimed to unravel fungal communities of natural O. sinensis using combination of high-throughput sequencing and culture-dependent approaches. RESULTS: A total of 280,519 high-quality sequences, belonging to 5 fungal phyla, 15 classes, 41 orders, 79 families, 112 genera, and 352 putative operational taxonomic units (OTUs) were obtained from natural O. sinensis using high-throughput sequencing. Among of which, 43 genera were identified in external mycelial cortices, Ophiocordyceps, Sebacinia and Archaeorhizomyces were predominant genera with the abundance of 95.86, 1.14, 0.85%, respectively. A total of 66 genera were identified from soil microhabitat, Inocybe, Archaeorhizomyces, unclassified Thelephoraceae, Tomentella, Thelephora, Sebacina, unclassified Ascomycota and unclassified fungi were predominant genera with an average abundance of 53.32, 8.69, 8.12, 8.12, 7.21, 4.6, 3.08 and 3.05%, respectively. The fungal communities in external mycelial cortices were significantly distinct from soil microhabitat. Meanwhile, seven types of culture media were used to isolate culturable fungi at 16 °C, resulted in 77 fungal strains identified by rDNA ITS sequence analysis, belonging to 33 genera, including Ophiocordyceps, Trichoderma, Cytospora, Truncatella, Dactylonectria, Isaria, Cephalosporium, Fusarium, Cosmospora and Paecilomyces, etc.. Among all culturable fungi, Mortierella and Trichoderma were predominant genera. CONCLUSIONS: The significantly differences and overlap in fungal community structure between two approaches highlight that the integration of high-throughput sequencing and culture-dependent approaches would generate more information. Our result reveal a comprehensive understanding of fungal community structure of natural O. sinensis, provide new insight into O. sinensis associated fungi, and support that microbiota of natural O. sinensis is an untapped source for novel bioactive metabolites discovery.

Diversity of filamentous fungi associated with coral and sponges in coastal reefs of northeast Brazil.

Fungi are known to form associations with various marine organisms and substrata such as sponges and corals, both as potential symbionts or pathogens. These microorganisms occupy an ecological niche that has recently attracted great attention due to their potential in either ecological or pharmaceutical advances. However, the interaction between marine invertebrates and fungi is still poorly understood, including how they are affected by anthropogenic actions. Here, we identified 89 fungal isolates through sequencing of the ITS rDNA region obtained from the various sponge and coral species collected at two northeast Brazilian reefs. We found 43 species of fungi from 16 genera, all belonging to phylum Ascomycota. The sponges and coral shared four genera: Aspergillus, Penicillium, Trichoderma, and Cladosporium, all commonly found in terrestrial habitats and associated with marine invertebrates. We observed some unusual species in relation to the marine environment, such as Clonostachys rosea and Neopestalotiopsis clavispora, most of them related to plants, either as saprophytic or pathogenic, suggesting that these species were transported from the surrounding terrestrial environment to the reefs. In addition, some isolates represent possible undescribed species, reinforcing the importance of studying the marine environment in relation to its ecological and biotechnological importance.

Predicting the concentration of indoor culturable fungi using a kernel-based extreme learning machine (K-ELM).

Indoor fungal is of great significance for human health. The kernel-based extreme learning machine is employed to determine the most important parameters for predicting the concentration of indoor culturable fungi (ICF). For model training and statistical analysis, parameters that contained indoor or outdoor PM10 and PM2.5, RH, Temperature, CO2 and ICF were measured in 85 residential buildings of Baoding, China, from November 2016 to March 2017. The variable selection process contains four different cases to identify the optimal input combination. The results indicate that root mean square error of the optimal input combinations can be improved 5.6% from 1 to 2 input variables, while that could be only improved 1.9% from 2 to 3 input variables. However, considering both precision and simplicity, the combination of indoor PM10 and RH provides a more suitable selection for predicting the ICF.

[Diversity of culturable fungi isolated from Coptis chinensis rot root in Shizhu city of Chongqing].

Root rot disease restricts the Coptis chinensis industry in Shizhu of Sichuan province. To disclose fungi composition and pathogen in rot root,so as to prevent and treat the root rot disease,the C. chinensis rot root of 5 years from 4 areas in Shizhu were collected in 3 seasons respectively. The fungi were isolated and molecularly and morphological identified,followed with population statistics. 437 fungi were isolated,belonging to 5 subphylum,11 classes,16 orders,22 families and 28 genus respectively. There are great difference among the fungi compositions of different area,year and sampling season,while there was no obvious variation rule. Ilyonectria sp.,Pythium sp.,Phoma sp,Trichoderma sp.are dominant genus,while Pythium sp.,Ilyonectria sp.,Phoma sp.,Fusarium sp. may contain root rot pathogen. Antagonistic bacteria may be screened from the strains of Trichoderma sp. isolated.

Isolation and identification of culturable fungi from the genitals and semen of healthy giant pandas (Ailuropoda melanoleuca).

BACKGROUND: In order to better understand the possible role of fungi in giant panda reproduction and overall health, it is important to provide a baseline for the normal fungal composition in the reproductive system. Using morphology and internal transcribed spacer (ITS) sequence analysis, we systematically isolated and identified fungal species from the vagina, foreskin, and semen of 21 (11 males and 10 females) healthy giant pandas to understand the normal fungal flora of the genital tracts. RESULTS: A total of 76 fungal strains were obtained, representing 42 genera and 60 species. Among them 47 fungal strains were obtained from vaginal samples, 24 from foreskins, and 5 from semen samples. Several fungal strains were isolated from more than one sample. More fungal species were isolated from females from males. The predominant genera were Aspergillus, Trichosporon, and Penicillium, followed by Candida, Cladosporium, Sordariomycetes, and Diaporthe. The average number of strains in the female vagina was significantly higher than in the foreskin and semen of male. CONCLUSIONS: A total of 60 fungal species (belonging to 42 genera) were identified in the giant panda's genital tract. Some of the species were commonly shared in both males and females. These findings provide novel information on the fungal community in the reproductive tracts of giant pandas.

Culturable fungi in potting soils and compost.

In the present study the spectrum and the incidence of fungi in potting soils and compost was investigated. Since soil is one of the most important biotopes for fungi, relatively high concentrations of fungal propagules are to be expected. For detection of fungi, samples of commercial soils, compost and soils from potted plants (both surface and sub-surface) were suspended and plated onto several mycological media. The resulting colonies were evaluated qualitatively and quantitatively. The results from the different sampling series vary, but concentrations on the surface of potted plants and in commercial soils are increased tenfold compared to compost and sub-surface soils. Median values range from 9.5 × 10(4) colony forming units (CFU)/g to 5.5 × 10(5) CFU/g. The spectrum of fungi also varies in the soils. However, all sampling series show high proportion of Aspergillus and Penicillium species, including potentially pathogenic species such as Aspergillus fumigatus. Cladosporium, a genus dominant in the ambient air, was found preferably in samples which were in contact with the air. The results show that potentially pathogenic fungi are present in soils. Immunocompromised individuals should avoid handling soils or potted plants in their immediate vicinity.

Household indoor air quality and its associations with childhood asthma in Shanghai, China: On-site inspected methods and preliminary results.

Few studies were conducted for associations of home environment with childhood health by on-site inspection in China. During 2013-2014, we conducted a case-control study with home inspection among 454 children (186 asthma children and 268 non-asthma children) in Shanghai, China. In this paper, we detailedly described the inspected methods and analyzed the preliminarily collected data. Except in winter, most residences meet the national standard for indoor temperature and relative humidity. Most living rooms had ≤1000ppm CO2, whereas over half of the child's bedrooms had slightly >1000ppm CO2 during night. Most residences had notably lower than 2500cfu/m3 airborne culturable fungi and ≤100µg/m3 formaldehyde. More than 70% of the child's bedrooms had ≤75µg/m3 PM2.5 and ≤150µg/m3 PM10. Indoor and outdoor concentrations of particulate matters had strong linear correlations (r=0.891-0.922; p-value <0.001). Most differences between cases and controls were not significant with respect to CO2, culturable fungi, formaldehyde, and particulate matters. Before and after adjusted for potential confounders, indoor averaged concentration of CO2 and particulate matters generally had negative associations with childhood history of doctor-diagnosed asthma in spring, summer, and autumn. Only in winter, indoor CO2 concentration was significantly associated with the increased odds of childhood asthma. Our results indicated that air quality among most residences in Shanghai could meet the national standard for indoor air quality in warm seasons; but household air quality and ventilation status in winter should be greatly improved. We suspected that those "unexpected" negative associations could exist due to changes in lifestyle behaviors regarding indoor air quality after the child being diagnosed asthma by a doctor.

Phylogenetic analyses and nitrate-reducing activity of fungal cultures isolated from the permanent, oceanic oxygen minimum zone of the Arabian Sea.

Reports on the active role of fungi as denitrifiers in terrestrial ecosystems have stimulated an interest in the study of the role of fungi in oxygen-deficient marine systems. In this study, the culturable diversity of fungi was investigated from 4 stations within the permanent, oceanic, oxygen minimum zone of the Arabian Sea. The isolated cultures grouped within the 2 major fungal phyla Ascomycota and Basidiomycota; diversity estimates in the stations sampled indicated that the diversity of the oxygen-depleted environments is less than that of mangrove regions and deep-sea habitats. Phylogenetic analyses of 18S rRNA sequences revealed a few divergent isolates that clustered with environmental sequences previously obtained by others. This is significant, as these isolates represent phylotypes that so far were known only from metagenomic studies and are of phylogenetic importance. Nitrate reduction activity, the first step in the denitrification process, was recorded for isolates under simulated anoxic, deep-sea conditions showing ecological significance of fungi in the oxygen-depleted habitats. This report increases our understanding of fungal diversity in unique, poorly studied habitats and underlines the importance of fungi in the oxygen-depleted environments.

Continued Organic Fertigation after Basal Manure Application Does Not Impact Soil Fungal Communities, Tomato Yield or Soil Fertility.

There is currently a limited understanding of the complex response of fungal microbiota diversity to organic fertigation. In this work, a 2-year field trial with organic tomato crops in a soil previously amended with fresh sheep manure was conducted. Two hypotheses were compared: (i) fertigation with organic liquid fertilizers versus (ii) irrigation with water. At the end of both years, soils were analyzed for physical-chemical parameters and mycobiome variables. Plate culture and DNA metabarcoding methods were performed in order to obtain a detailed understanding of soil fungal communities. Fertigation did not increase any of the physical-chemical parameters. Concerning soil fungal communities, differences were only found regarding the identification of biomarkers. The class Leotiomycetes and the family Myxotrichaceae were identified as biomarkers in the soil fungal community analyzed by means of DNA metabarcoding of the "fertigation" treatment at the end of Year 1. The Mortierella genus was detected as a biomarker in the "water" treatment, and Mucor was identified in the "fertigation" treatment in the cultivable soil fungi at the end of Year 2. In both years, tomato yield and fruit quality did not consistently differ between treatments, despite the high cost of the fertilizers added through fertigation.

The First Comprehensive Biodiversity Study of Culturable Fungal Communities Inhabiting Cryoconite Holes in the Werenskiold Glacier on Spitsbergen (Svalbard Archipelago, Arctic).

Cryoconite holes on glacier surfaces are a source of cold-adapted microorganisms, but little is known about their fungal inhabitants. Here, we provide the first report of distinctive fungal communities in cryoconite holes in the Werenskiold Glacier on Spitsbergen (Svalbard Archipelago, Arctic). Due to a combination of two incubation temperatures (7 °C and 24 ± 0.5 °C) and two media during isolation (PDA, YPG), as well as classical and molecular identification approaches, we were able to identify 23 different fungi (21 species and 2 unassigned species). Most of the fungi cultured from cryoconite sediment were ascomycetous filamentous micromycetes. However, four representatives of macromycetes were also identified (Bjerkandera adusta, Holwaya mucida, Orbiliaceae sp., and Trametes versicolor). Some of the described fungi possess biotechnological potential (Aspergillus pseudoglaucus, A. sydowii, Penicillium expansum, P. velutinum, B. adusta, and T. versicolor), thus, we propose the Arctic region as a source of new strains for industrial applications. In addition, two phytopathogenic representatives were present (P. sumatraense, Botrytis cinerea), as well as one potentially harmful to humans (Cladosporium cladosporioides). To the best of our knowledge, we are the first to report the occurrence of A. pseudoglaucus, C. allicinum, C. ramotenellum, P. sumatraense, P. velutinum, P. cumulodentata, B. adusta, and T. versicolor in polar regions. In all likelihood, two unassigned fungus species (Orbiliaceae and Dothideomycetes spp.) might also be newly described in such environments. Additionally, due to experimenting with 10 sampling sites located at different latitudes, we were able to conclude that the number of fungal spores decreases as one moves down the glacier. Considering the prevalence and endangerment of glacial environments worldwide, such findings suggest their potential as reservoirs of fungal diversity, which should not be overlooked.

Diversity and Metabolic Activity of Fungi Causing Biodeterioration of Canvas Paintings.

Research into the biodeteriorative potential of fungi can serve as an indicator of the condition of heritage items. Biodeterioration of canvas paintings as a result of fungal metabolic activity is understudied with respect to both the species diversity and mechanisms involved. This study brings new evidence for the physiology of fungi biodeteriorative capacity of canvas paintings. Twenty-one fungal isolates were recovered from four oil paintings (The Art Museum, Cluj-Napoca) and one gouache painting (private collection), dating from the 18th to 20th centuries. The species, identified based on the molecular markers Internal Transcribed Spacer (ITS), beta-tubulin (tub2), or translation elongation factor 1 (TEF-1), are common colonisers of canvas paintings or indoor environments (e.g., Penicillium spp., Aspergillus spp., Alternaria spp.). Fungi enzymatic profiles were investigated by means of hydrolysable substrates, included in culture media or in test strips, containing components commonly used in canvas paintings. The pigment solubilisation capacity was assessed in culture media for the primary pigments and studied in relation to the organic acid secretion. Caseinases, amylases, gelatinases, acid phosphatase, N-acetyl-ß-glucosaminidase, naphthol-AS-BI-phosphohydrolase, and ß-glucosidase were found to be the enzymes most likely involved in the processes of substrate colonisation and breakdown of its components. Aureobasidium genus was found to hold the strongest biodeteriorative potential, followed by Cladosporium, Penicillium, Trichoderma, and Aspergillus. Blue pigment solubilisation was detected, occurring as a result of organic acids secretion. Distinct clusters were delineated considering the metabolic activities detected, indicating that fungi specialise in utilisation of certain types of substrates. It was found that both aged and modern artworks are at risk of fungal biodeterioration, due to the enzymatic activities' diversity and intensity, pigment solubilisation capacity or pigment secretion.

Diversity, Abundance, and Ecological Roles of Planktonic Fungi in Marine Environments.

Fungi are considered terrestrial and oceans are a "fungal desert". However, with the considerable progress made over past decades, fungi have emerged as morphologically, phylogenetically, and functionally diverse components of the marine water column. Although their communities are influenced by a plethora of environmental factors, the most influential include salinity, temperature, nutrients, and dissolved oxygen, suggesting that fungi respond to local environmental gradients. The biomass carbon of planktonic fungi exhibits spatiotemporal dynamics and can reach up to 1 µg CL-1 of seawater, rivaling bacteria on some occasions, which suggests their active and important role in the water column. In the nutrient-rich coastal water column, there is increasing evidence for their contribution to biogeochemical cycling and food web dynamics on account of their saprotrophic, parasitic, hyper-parasitic, and pathogenic attributes. Conversely, relatively little is known about their function in the open-ocean water column. Interestingly, methodological advances in sequencing and omics approach, the standardization of sequence data analysis tools, and integration of data through network analyses are enhancing our current understanding of the ecological roles of these multifarious and enigmatic members of the marine water column. This review summarizes the current knowledge of the diversity and abundance of planktonic fungi in the world's oceans and provides an integrated and holistic view of their ecological roles.

Diversity and Antiaflatoxigenic Activities of Culturable Filamentous Fungi from Deep-Sea Sediments of the South Atlantic Ocean.

Despite recent studies, relatively few are known about the diversity of fungal communities in the deep Atlantic Ocean. In this study, we investigated the diversity of fungal communities in 15 different deep-sea sediments from the South Atlantic Ocean with a culture-dependent approach followed by phylogenetic analysis of ITS sequences. A total of 29 fungal strains were isolated from the 15 deep-sea sediments. These strains belong to four fungal genera, including Aspergillus, Cladosporium, Penicillium, and Alternaria. Penicillium, accounting for 44.8% of the total fungal isolates, was a dominant genus. The antiaflatoxigenic activity of these deep-sea fungal isolates was studied. Surprisingly, most of the strains showed moderate to strong antiaflatoxigenic activity. Four isolates, belonging to species of Penicillium polonicum, Penicillium chrysogenum, Aspergillus versicolor, and Cladosporium cladosporioides, could completely inhibit not only the mycelial growth of Aspergillus parasiticus mutant strain NFRI-95, but also the aflatoxin production. To our knowledge, this is the first report to investigate the antiaflatoxigenic activity of culturable deep-sea fungi. Our results provide new insights into the community composition of fungi in the deep South Atlantic Ocean. The high proportion of strains that displayed antiaflatoxigenic activity demonstrates that deep-sea fungi from the Atlantic Ocean are valuable resources for mining bioactive compounds.

Residential airborne culturable fungi under general living scenario: On-site investigation in 12 typical cities, China.

BACKGROUND: Residential airborne fungi may present obvious risk to human health. However, many countries do not recognize the necessarily need to control residential airborne culturable fungi (RAF). In China, few systemic investigations have been conducted to illustrate the distribution of residential airborne fungi and identify the association between indoor influencing variables and RAF under general living scenario in China. OBJECTIVE: This study aimed to investigate RAF with the on-site research of 12 typical cities in China, and provided the latest characteristics and potential influencing factors of RAF under general living scenario. METHODS: We measured RAF and investigated residential characteristics in 12 typical cities in China, 2018. At least 50 resident families were randomly selected both from downwind and upwind districts in each city with pre-proposed requirements. The RAF were sampled by the six-stage Anderson impactor. PM2.5 and PM10 were monitored by calibrated light-scattering dust meters. CO and CO2 were monitored by non-dispersive infrared analyzer method. NO2 was determined by Saltzman method. General linear model was used to evaluate the association between RAF exposure and residential characteristics with adjustment for potential confounders. RESULTS: The RAF concentrations ranged from 0 to 9371 CFU/m3 with a median concentration of 396 CFU/m3. The median concentrations of RAF in the warm season were statistically higher than the cold season in Panjin, Qingdao, Lanzhou and Luoyang, but lower than the cold season in Shijiazhuang, Ningbo and Nanning. RAF in the bedrooms were more than the living rooms in all cities except Xi'an. Temperature and humidity had an interactive effect on the RAF (OR = 1.0006, 95% CI: 1.0005, 1.0006). Some residential environmental pollutants, including PM2.5 (OR = 0.9989, 95% CI: 0.9988, 0.9989), PM10 (OR = 0.9993, 95% CI: 0.9993, 0.9993), and CO2 (OR = 0.0236, 95% CI: 0.0230, 0.0243), were negatively correlated with RAF. CO (OR = 1.1450, 95% CI: 1.1433, 1.1467) and NO2 (OR = 1.0026, 95% CI: 1.0024, 1.0028) were positively correlated with RAF. Architectural characteristics (sunlight exposure, building history, longitude, latitude, total living area, living floor, distance from the road, house type, the layers of window glass and decoration), family-related information (income) and lifestyle behaviors (keeping pets, growing plants, cooking, using insecticide, burning incense, heating, using air conditioner and cleaning frequencies) were also significantly related with RAF. CONCLUSIONS: This study reported nation-wide baseline condition of RAF and related influencing factors under general living scenario with quantitative details, which are exceedingly promising for evidence-driven standard and reasonable control strategy of residential airborne culturable in China.

Exploring the potential relationship between indoor air quality and the concentration of airborne culturable fungi: a combined experimental and neural network modeling study.

Indoor airborne culturable fungi exposure has been closely linked to occupants' health. However, conventional measurement of indoor airborne fungal concentration is complicated and usually requires around one week for fungi incubation in laboratory. To provide an ultra-fast solution, here, for the first time, a knowledge-based machine learning model is developed with the inputs of indoor air quality data for estimating the concentration of indoor airborne culturable fungi. To construct a database for statistical analysis and model training, 249 data groups of air quality indicators (concentration of indoor airborne culturable fungi, indoor/outdoor PM2.5 and PM10 concentrations, indoor temperature, indoor relative humidity, and indoor CO2 concentration) were measured from 85 residential buildings of Baoding (China) during the period of 2016.11.15-2017.03.15. Our results show that artificial neural network (ANN) with one hidden layer has good prediction performances, compared to a support vector machine (SVM). With the tolerance of ± 30%, the prediction accuracy of the ANN model with ten hidden nodes can at highest reach 83.33% in the testing set. Most importantly, we here provide a quick method for estimating the concentration of indoor airborne fungi that can be applied to real-time evaluation.

Phylogenetic diversity of culturable fungi in the Heshang Cave, central China.

Caves are nutrient-limited and dark subterranean ecosystems. To date, attention has been focused on geological research of caves in China, whilst indigenous microbial diversity has been insufficiently characterized. Here, we report the fungal diversity in the pristine, oligotrophic, karst Heshang Cave, central China, using a culture-dependent method coupled with the analysis of the fungal rRNA-ITS gene sequences. A total of 194 isolates were obtained with six different media from 14 sampling sites of sediments, weathered rocks, and bat guanos. Phylogenetic analysis clustered the 194 sequenced isolates into 33 genera within 15 orders of three phyla, Ascomycota, Basidiomycota, and Zygomycota, indicating a high degree of fungal diversity in the Heshang Cave. Notably, 16 out of the 36 fungal genera were also frequently observed in solution caves around the world and 23 genera were previously found in carbonate cave, indicating potential similarities among fungal communities in cave ecosystems. However, 10 genera in this study were not reported previously in any solution caves, thus expanding our knowledge about fungal diversity in cave ecosystems. Moreover, culturable fungal diversity varied from one habitat to another within the cave, being the highest in sediments, followed by weathered rocks and bat guanos as indicated by α-diversity indexes. At the genus level, Penicillium accounted for 40, 54, and 52% in three habitats of sediments, weathered rocks, and bat guanos, respectively. Trichoderma, Paecilomyces, and Aspergillus accounted for 9, 22, and 37% in the above habitats, correspondingly. Despite of the dominance of Penicillium in all samples, ß-diversity index indicated significant differences between each two fungal communities in the three habitats in view of both the composition and abundance. Our study is the first report on fungal communities in a natural pristine solution cave system in central China and sheds light on fungal diversity and functions in cave ecosystems.