Anna Maria Picco

JEM Spotlight: Fungi, mycotoxins and microbial volatile organic compounds in mouldy interiors from water-damaged buildings

Concerns have been raised about exposure to mycotoxin producing fungi and the microbial volatile organic compounds (MVOCs) they produce in indoor environments. Therefore, the presence of fungi and mycotoxins was investigated in 99 samples (air, dust, wallpaper, mycelium or silicone) collected in the mouldy interiors of seven water-damaged buildings. In addition, volatile organic compounds (VOCs) were sampled. The mycotoxins were analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (20 target mycotoxins) and quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). Morphological and molecular identifications of fungi were performed. Of the 99 samples analysed, the presence of one or more mycotoxins was shown in 62 samples by means of LC-MS/MS analysis. The mycotoxins found were mainly roquefortine C, chaetoglobosin A and sterigmatocystin but also roridin E, ochratoxin A, aflatoxin B(1) and aflatoxin B(2) were detected. Q-TOF-MS analysis elucidated the possible occurrence of another 42 different fungal metabolites. In general, the fungi identified matched well with the mycotoxins detected. The most common fungal species found were Penicillium chrysogenum, Aspergillus versicolor (group), Chaetomium spp. and Cladosporium spp. In addition, one hundred and seventeen (M)VOCs were identified, especially linear alkanes (C(9)-C(17)), aldehydes, aromatic compounds and monoterpenes.

Assessment of airborne microorganism contamination in an industrial area characterized by an open composting facility and a wastewater treatment plant

In order to assess the potential exposure hazard to workers and people living in the immediate surroundings of an area characterized by an open composting facility and a wastewater treatment plant, a quantitative and a qualitative analysis of airborne microorganisms were carried out. Air sampling was performed once a week for four consecutive weeks in summer and winter. Six sites were selected as air sampling sites: one was upwind at approximately 40 m from the facilities; the other five were downwind at increasing distances from the facilities, with the furthest at 100 m away. Monitoring permitted us to verify the influence that the composting activities and wastewater treatment had on the bacterial and fungal contamination of the air. The results obtained have been expressed by means of contamination indexes that have already been used in previous works: a major microbiological contamination near the plants was evidenced. Near the facilities, mesophilic bacteria, psychrophilic bacteria and microfungi showed the highest median concentrations, respectively, of 307.5, 327.5 and 257.5 CFU/m(3). Moreover, the season generally influenced the concentration of the bacteria as well as of the fungi; higher in summer than in winter. The contamination index global index of microbial contamination (GIMC/m(3)) showed mean values of 4058.9 in summer and 439.7 in winter and the contamination index-amplification index (AI) showed values of 4.5 and 1.1 in the same seasons, respectively. Controlling the seasonal effect, mesophilic bacteria, Pseudomonas spp. and Enterobacteriaceae showed a significant decline in concentration with respect to upwind air samples and with increasing distance. Both GIMC and AI showed a significant decline with respect to upwind air samples by increasing the distance from facilities after adjusting for the seasonal effect. In conclusion, even if these plants do not represent a potential risk for nearby populations, they may pose a potential health risk for workers.

Determination of aerial microbiological contamination in scholastic sports environments

Aims: To assess the microbiological indoor air quality (IAQ) of high school and college gyms during physical training lessons and to evaluate the effective microbiological exposure of students.

Identification of volatile markers for indoor fungal growth and chemotaxonomic classification of Aspergillus species.

Microbial volatile organic compounds (MVOCs) were collected in water-damaged buildings to evaluate their use as possible indicators of indoor fungal growth. Fungal species isolated from contaminated buildings were screened for MVOC production on malt extract agar by means of headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (GC-MS) analysis. Some sesquiterpenes, specifically derived from fungal growth, were detected in the sampled environments and the corresponding fungal producers were identified. Statistical analysis of the detected MVOC profiles allowed the identification of species-specific MVOCs or MVOC patterns for Aspergillus versicolor group, Aspergillus ustus, and Eurotium amstelodami. In addition, Chaetomium spp. and Epicoccum spp. were clearly differentiated by their volatile production from a group of 76 fungal strains belonging to different genera. These results are useful in the chemotaxonomic discrimination of fungal species, in aid to the classical morphological and molecular identification techniques.

Assessment of genetic diversity in Italian rice germplasm related to agronomic traits and blast resistance (Magnaportheoryzae)

Italy is the only country in Europe with a significant land area used for rice production. In this paper, the genetic diversity of 172 national varieties and 47 foreign accessions (ITALORYZA collection) was investigated using a set of neutral markers evenly distributed throughout the 12 chromosomes. Out of the 218 alleles detected in our analysis, 17 and 29% were specific to the Italian and foreign accessions, respectively. From the neighbour-joining tree generated, six sub-groups of temperate japonica germplasm were identified. Plant height and grain type measured in the source collection were fitted to the phylogenetic tree, along with the period of variety registration. This integrated genotype–phenotype analysis revealed that specific sub-groups are characterized by uniform classes of grain type, or by similar plant size, or by period of release in the market. The whole collection was also evaluated for leaf blast resistance by inoculating the plants with three strains of Magnaporthe oryzae, representing the pathogen genetic diversity existing in Italy. Only 15 out of 172 Italian accessions (8.7%) were resistant to all three fungal strains. The correlation between genotype and leaf blast phenotype revealed that the most highly resistant Italian varieties are included in a single germplasm sub-group derived from US varieties. This study represents the starting point for carrying out detailed phenotype–genotype whole-genome association studies and identification of the genetic basis of important agronomic traits for rice cultivation in temperate climates.

Airborne fungi as biocontaminants at two Milan underground stations

The viable or culturable air spora of two Milan underground stations and of the squares above has been investigated during a one-year study. Aerobiological data were collected by semi-quantitative method. Outdoor results show the presence of four dominant genera, Cladosporium, Penicillium, Epicoccum and Alternaria, whose presence varied throughout the year. Cryptococcus neoformans was isolated in Duomo Square. Indoor sampling shows to what extent the mycoflora of an indoor environment can depend both on the fungal spora coming from outside and the capacity of the fungi to colonize the different sublayers found indoors.

Functional Analysis of the Degradation of Cellulosic Substrates by a Chaetomium globosum Endophytic Isolate

ABSTRACT Most photosynthetically fixed carbon is contained in cell wall polymers present in plant biomasses, the largest organic carbon source in the biosphere. The degradation of these polymers for biotechnological purposes requires the combined action of several enzymes. To identify new activities, we examined which enzymes are activated by an endophytic strain of Chaetomium globosum to degrade cellulose-containing substrates. After biochemical analyses of the secretome of the fungus grown on cellulose or woody substrates, we took advantage of the available genomic data to identify potentially involved genes. After in silico identification of putative genes encoding either proteins able to bind to cellulose or glycohydrolases (GHs) of family 7, we investigated their transcript levels by reverse transcription-quantitative PCR (RT-qPCR). Our data suggest that eight genes compose the core of the cellulose-degrading system of C. globosum. Notably, the related enzymes belong structurally to the well-described GH families 5, 6, 7, 16, and 45, which are known to be the core of the cellulose degradation systems of several ascomycetes. The high expression levels of cellobiose dehydrogenase and two GH 61 enzymes suggest the involvement of this oxidoreductive synergic system in C. globosum. Transcript analysis along with relevant coding sequence (CDS) isolation and expression of recombinant proteins proved to be a key strategy for the determination of the features of two endoglucanases used by C. globosum for the first attack of crystalline cellulose. Finally, the possible involvement of transcriptional regulators described for other ascomycetes is discussed.

Superficial mycoses in Italy

A total of 1244 cases suspected of superficial mycoses were analysed in a period of 12 months in Italy. Dermatophytoses were found in 62.4%; the other common disease was Pityriasis versicolor (44.1%). The predominant speciesEpidermophyton floccosum (34,2%) was strickingly high in relation to other statistics; this species was commonest in tinea cruris. Among the other dermatophytesM. canis was predominant (31.3%) commonest in tinea corporis, followed byT. mentagrophytes (17,8%) andT. rubrum (10,3%). A high rate of yeasts was found in tinea manum (81.6%) and tinea pedis (47.7%).

Biodiversity of emerging pathogenic and invasive fungi in plants, animals and humans in Italy

Abstract In the last 30 years, the spread of emerging and invasive fungal pathogens has had a profound impact on plants, animals and humans, causing crop losses and damaging their health with economic and social impact. Awareness of the biodiversity of these pathogens is essential for their detection, identification and control. In this article, their diversity in Italy is reviewed taking into consideration the international context. Fungal endophytes, important ecological agents whose lifestyle can be switched by stochastic events from beneficial/neutral to pathogenic, are included in this review.

High spots for diversity of soil and litter microfungi in Italy

Abstract Soil fungal diversity plays a fundamental role in delivering key ecosystem goods and services. This article assesses diversity of saprobic soil and litter microfungi, as taxonomical and functional components which affect above- and below-ground relationships within Alpine and Mediterranean regions of Italy. We highlighted biodiversity high spots focusing on four research topics that have been developed over time and are currently in progress in Italy. Preliminary quantitative data concerning soil microfungi in the Raethian Alps showed a strong reduction of Colony Forming Unit (CFU)s with altitude. Keratinophilic microfungi in natural and anthropogenic environments were widespread among filamentous fungi and 121 species have been isolated in Italy since 1960. Heat stimulated microfungi in Mediterranean region soils showed high values both in abundance and species density even two years after the experimental fire, with Neosartorya spp. playing a pivotal role. The diversity of microfungi of Quercus ilex (150 species), in the Mediterranean region, higher than that in leaf litter of other species, was explained mainly by different forms of growth and the phytoclimatic characters of the areas under study.