Marinella Rodolfi

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.

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.

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.

Pyrogenic activity of air to characterize bioaerosol exposure in public buildings: a pilot study

Aims:  The aim of our study was to investigate indoor air quality (IAQ) by comparing pyrogen concentration and microbiological contamination in offices in public buildings.

Pyricularia grisea and Bipolaris oryzae: a preliminary study on the occurrence of airborne spores in a rice field

Qualitative and quantitative analysis of fungalair spores in a rice field located around Pavia(North Italy) were made from 10th June to7th October 1996. Quantitative data wereanalyzed for the two rice pathogensPyricularia grisea (Cooke) Sacc. andBipolaris oryzae Shoem., causal organisms ofblast and brown spot. Results showed thatB. oryzae was detected at the end of June,reaching its peak in July. Brown spot symptomsin-field were detected six-seven days after theaforementioned peak. Pyricularia griseawas monitored later than Bipolaris as itwas detected for the first time on July21st, reaching its peaks on July 27thand August 6–7th. Field evaluation of thedisease showed the presence of blast startingfrom the first week of July.The knowledge of the atmospheric concentrationof Pyricularia and Bipolarisairborne spores together with a correctexamination of the crops can yield informationabout the risk of infection during thevegetative season, thus allowing for a moreaccurate use of fungicides on rice crops,according to the modern conception ofintegrated control.

Differential responsesin vitroof rice cultivars to Italian lineages of the blast pathogenPyricularia grisea (Cooke) Sacc. 1. Oxidative burst

Suspension cultured cells of six rice cultivars differing in their sensitivity to blast were treated with mycelial wall hydrolysates prepared from seven isolates belonging to different Pyricularia grisea lineages. Soon after elicitor addition, rice cells produced significant amounts of superoxide anion, which was rapidly converted into diffusible peroxide. Maximal effects were achieved at 50 mg L-1 elicitor. In all cases, a 7 to 13-fold increase in the basal rate of reactive oxygen species production was found. Neither differential effects among strains nor clear relationships between lineage and the resulting oxidative burst were evident. Interestingly, a good correlation was found between basal (and elicited) levels of peroxide generation and the overall tolerance of rice cultivars to the pathogen. About two days after elicitation, cell death occurred proportional to the amount of hydrogen peroxide released. Peroxide was required to trigger loss of cell viability, but the latter was not due to a direct toxic effect, suggesting the induction of programmed cell death. Results represent the first data aimed to develop in vitro tests for pathogenicity prediction of Italian blast lineages toward rice cultivars.

Evaluation of microbiological air quality and of microclimate in university classrooms.

The proliferation of air-diffused microorganisms inside public buildings such as schools, hospitals, and universities, is often indicated as a possible health risk. In this research, we have illustrated the results of an investigation realized to determine the health of the air in some university classrooms, both from a microbiological and a microclimatic viewpoint, during the normal didactic activity of direct lessons. The results obtained have been expressed by means of contamination indices, already used in previous works. Very little contamination was recorded in the different phases of air treatment, which underlines the efficiency of the system and of the maintenance protocols. The Global Index of Microbial Contamination (GIMC per cubic meter) showed a value greater than the mean during the heating period (290), while the highest values (95th percentile 1,138.45) were recorded in the period using air conditioning. The index of mesophilic bacterial contamination, though it did not show any significant differences in the various modes of air treatment, showed a mean value (1.34) and the 95th percentile value (4.14), which was greater in the air-conditioning phase. Finally, the mean value of the amplification index underlined a decrease in the microbial contamination in comparison to the outside, while showing situations of increased microbial amplification during the period of simple ventilation (95th percentile 4.27). The 95th percentile values found for GICM in the three sampling periods, however, permitted us to identify the value of GIMC per cubic meter equal to 1,000 as a guide to provide a means of self-monitoring the quality of the air inside the classrooms. From a microclimatic viewpoint, two periods of the year manifested discomfort situations: during the heating phase (winter) and during the simple ventilation phase (spring). The results obtained indicate, therefore, a need to intervene on the environmental parameters, not being able, in this particular case, to intervene on other aspects that influence the microclimate.

Biochemistry of lipolytic enzymes secreted by Penicillium solitum and Cladosporium cladosporioides.

Two distinct extracellular lipases were obtained from Penicillium solitum 194A, isolated from domestic compost, and Cladosporium cladosporioides 194B, isolated from dairy wastewater. These alkaline enzymes had molecular masses of 42 and 30 kDa, respectively. The P. solitum 194A lipase differed in mass from previously reported enzyme, indicating that it is a novel lipase, and indicating that penicillia can secrete lipase isoenzymes. The C. cladosporioides lipase was more active on esters of medium-chain acids, whereas the P. solitum lipase was more active on longer chained substrates. The C. cladosporioides enzyme displayed higher thermal stability than the P. solitum lipase, preserving full activity up to 48 °C and showing a T50 (10 min) of 60 °C. Their different catalytic properties and good protein stability should make these enzymes suitable for biotechnological applications. Furthermore, the combined use of these two fungal strains may prove to be valuable in lipid-rich waste management.