Characterization of the bioaerosol in a natural thermal cave and assessment of the risk of transmission of SARS-CoV-2 virus.

Thermal caves represent an environment characterized by unique chemical/physical properties, often used for treatment and care of musculoskeletal, respiratory, and skin diseases.However, these environments are poorly characterized for their physical and microbiological characteristics; furthermore, the recent pandemic caused by COVID-19 has highlighted the need to investigate the potential transmission scenario of SARS-CoV-2 virus in indoor environments where an in-depth analysis of the aerosol concentrations and dimensional distributions are essential to monitor the spread of the virus.This research work was carried out inside a natural cave located in Viterbo (Terme dei Papi, Italy) where a waterfall of sulfur-sulfate-bicarbonate-alkaline earth mineral thermal water creates a warm-humid environment with 100% humidity and 48 °C temperature. Characterization of the aerosol and bioaerosol was carried out to estimate the personal exposure to aerosol concentrations, as well as particle size distributions, and to give an indication of the native microbial load.The data obtained showed a predominance of particles with a diameter greater than 8 µm, associated with low ability of penetration in the human respiratory system. A low microbial load was also observed, with a prevalence of noncultivable strains generated by the aerosolization of the thermal waters.Finally, the estimation of SARS-CoV-2 infection risk by means of mathematical modeling revealed a low risk of transmission, with a decisive effect given by the mechanical ventilation system, which together with the adoption of social distancing measures makes the risk of infection extremely low.

Orange peel waste-based liquid medium for biodiesel production by oleaginous yeasts.

Orange peel waste (OPW), the primary byproduct of the juice extraction process, is annually generated in massive amounts (21 Mton), and its aqueous extraction in biorefining operations yields a liquid fraction, referred to as orange peel extract (OPE). Although OPE contains significant amounts of easily assimilable carbohydrates, such as fructose, glucose, and sucrose, no investigations have been conducted yet to assess its possible use in biodiesel production by oleaginous yeasts. Consequently, the objective of the present study was to assess whether OPE might act as the basis of a liquid medium for microbial lipid production. A screening conducted with 18 strains of oleaginous yeasts in shaken flask on the OPE-based medium showed that Rhodosporidium toruloides NRRL 1091 and Cryptococcus laurentii UCD 68-201 gave the best results in terms of lipid production (5.8 and 4.5 g L-1, respectively) and accumulation (77 and 47% on a dry matter basis, respectively). The subsequent scale transfer of the process to a 3-L STR operated in batch mode halved the time required to reach the lipid peak with the ensuing increase in volumetric productivities in R. toruloides NRRL 1091 (3646 mg L-1 day-1) and C. laurentii UCD 68-201 (2970.7 mg L-1 day-1). The biodiesel yields from the lipids of the former and the latter strain were 36.9 and 31.9%, respectively. Based on multivariate analysis of fatty acid methyl ester compositions, the lipids from the former and the latter strain were highly resembling those of Jatropha and palm oils, two commonly used feedstocks for biodiesel manufacturing.

Meteorite-catalyzed syntheses of nucleosides and of other prebiotic compounds from formamide under proton irradiation.

Liquid formamide has been irradiated by high-energy proton beams in the presence of powdered meteorites, and the products of the catalyzed resulting syntheses were analyzed by mass spectrometry. Relative to the controls (no radiation, or no formamide, or no catalyst), an extremely rich, variegate, and prebiotically relevant panel of compounds was observed. The meteorites tested were representative of the four major classes: iron, stony iron, chondrites, and achondrites. The products obtained were amino acids, carboxylic acids, nucleobases, sugars, and, most notably, four nucleosides: cytidine, uridine, adenosine, and thymidine. In accordance with theoretical studies, the detection of HCN oligomers suggests the occurrence of mechanisms based on the generation of radical cyanide species (CN·) for the synthesis of nucleobases. Given that many of the compounds obtained are key components of extant organisms, these observations contribute to outline plausible exogenous high-energy-based prebiotic scenarios and their possible boundary conditions, as discussed.

First Evidence on the Role of Heavy Ion Irradiation of Meteorites and Formamide in the Origin of Biomolecules.

Formamide (NH2CHO) has been irradiated in condensed phase at 273 K by 11B-boron beams in the presence of powdered meteorites of the chondrite and stony-iron types. Relative to the controls (no radiation or no catalysis), a variegate panel of compounds was observed, including purine and pyrimidine nucleobases (uracil, cytosine, adenine, and guanine), nucleobase analogues, heterocycles, and carboxylic acids involved in metabolic pathways. The presence of amino imidazole carbonitrile (AICN), 4,6-diamino purine (4,6-DAP) and 2,4-diamino pyrimidine (2,4-DAPy) among the observed products suggests the occurrence of an unified mechanism based on the generation of radical cyanide species (•CN). These observations contribute to outline plausible prebiotic scenarios involving 11B-boron as energy source.

Valorization of winery and distillery by-products by hydrothermal carbonization.

This work aims at finding an alternative strategy to manage the waste generated by the winemaking industry to obtain a solid biofuel and phenolic compounds. The effect of temperature (180-260 °C), residence time (1-7 h), and biomass-to-liquid ratio (0.05-0.25) on the co-hydrothermal carbonization of vine pruning and exhausted grape pomace, by using vinasse as moisture source, is studied. The effect of the variables is investigated and optimized using the Box-Behnken design of response surface methodology to maximize mass yield, fuel ratio, energy densification yield and phenols extraction yield and to minimize energy consumption. The statistical analysis shows that the carbonization temperature is a crucial parameter of the process, decreasing the product yield on one hand and improving the quality of hydrochar on the other. At the optimal conditions (246.3 °C, 1.6 h, 0.066), an hydrochar yield of 52.64% and a calorific value of 24.1 MJ/kg were obtained. Moreover, the analysis of the H/C and O/C ratios of hydrochars demonstrates that carbonisation significantly improves the fuel properties of solid biofuel. Liquid by-products obtained from the HTC process are found to contain high concentrations of organic matter but the BOD/COD ratios suggest their potential valorization by biological methods.

Mixed glycerol and orange peel-based substrate for fed-batch microbial biodiesel production.

The aqueous extraction of orange peel waste (OPW), the byproduct of the juice extraction process generated annually in massive amounts (21 Mton), yields a carbohydrate-rich liquid fraction, termed orange peel extract (OPE). Several studies highlight that the combination of glycerol, a biodiesel byproduct, with carbohydrate mixtures might boost microbial lipid production. This study performed first a shaken flask screening of 15 oleaginous yeast strains based on their growth and lipid-producing abilities on OPE- and glycerol-based media. This screening enabled the selection of R. toruloides NRRL 1091 for the assessment of the process transfer in a stirred tank reactor (STR). This assessment relied, in particular, on either single- and double-stage feeding fed-batch (SSF-FB and DSF-FB, respectively) processes where OPE served as the primary medium and nitrogen-containing glycerol-OPE mixtures as the feeding one. The continuous supply mode at low dilution rates (0.02 and 0.01 h-1 for SSF-FB and DSF-FB, respectively) starting from the end of the exponential growth of the initial batch phase enabled the temporal extension of biomass and lipid production. The SSF-FB and DSF-FB processes attained high biomass and lipid volumetric productions (LVP) and ensured significant lipid accumulation on a dry cell basis (YL/X). The SSF-FB process led to LVP of 20.6 g L-1 after 104 h with volumetric productivity (r L) of 0.20 g L-1 h-1 and YL/X of 0.80; the DSF-FB process yielded LVP, r L and YL/X values equal to 15.92 g L-1, 0.11 g L-1 h-1 and 0.65, respectively. The fatty acid profiles of lipids from both fed-batch processes were not significantly different and resembled that of Jatropha oil, a vastly used feedstock for biodiesel production. These results suggest that OPE constitutes an excellent basis for the fed-batch production of R. toruloides lipids, and this process might afford a further option in OPW-based biorefinery.

Production of lignin-modifying enzymes by Trametes ochracea on high-molecular weight fraction of olive mill wastewater, a byproduct of olive oil biorefinery.

The high-molecular weight fraction of olive mill wastewater (HMW-OMW), a byproduct of olive oil biorefinery, was used at the reactor level as the basal medium for production of laccase and Mn-dependent peroxidase (MnP) by Trametes ochracea. Three reactor systems, namely stirred tank reactors equipped with either Rushton turbines or marine impeller and draft tube (STR and STR-MD, respectively) and an air-lift reactor (ALR) were compared for this purpose. Although inocula were supplied as intact pellets, in both STR-based systems fungal growth evolved rapidly into a dispersed form while the ALR enabled the maintenance of the pellet growth mode. STR was deemed to be the most promising system since it best supported the production MnP activity on the HMW-OMW-based medium and its performance in laccase production did not differ from that observed with the STR-MD. Among the stirring regimes considered (250, 400, 500 and 600 rpm), the best production in the STR was observed at 500 rpm and 1.0 vvm for both laccase (8850 ± 270 IU L-1 on day 15) and MnP (17,027.4 ± 87.2 IU L-1 on day 13). When the inocula were supplied to the STR in homogenized form, the MnP production peak (16,856 ± 1070 IU L-1) was attained 8 days earlier than the previous condition and that of laccase was nearly doubled (14,967 ± 907 IU L-1). When compared with literature data, T. ochracea MnP production and productivity on the HMW-OMW-based medium were the highest reported for a wild-type fungal strain.

A sustainable use of Ricotta Cheese Whey for microbial biodiesel production.

The increasing demand of plant oils for biodiesel production has highlighted the need for alternative strategies based either on non-food crops or agro-industrial wastes that do not compete with food and feed production. In this context, the combined use of wastewater and oleaginous microorganisms could be a valuable production option. Ricotta cheese whey (RCW), one of the major byproducts of the dairy industry, is produced in very high and steadily increasing amounts and, due to its high organic load, its disposal is cost-prohibitive. In the present study, in order to assess the adequacy of RCW as a growth medium for lipid production, 18 strains of oleaginous yeasts were investigated in shaken flask for their growth and lipid-producing capabilities on this substrate. Among them, Cryptococcus curvatus NRRL Y-1511 and Cryptococcus laurentii UCD 68-201 adequately grew therein producing substantial amounts of lipids (6.8 and 5.1gL-1, respectively). A high similarity between the percent fatty acid methyl esters (FAME) composition of lipids from the former and the latter strain was found with a predominance of oleic acid (52.8 vs. 48.7%) and of total saturated fatty acids (37.9 vs. 40.8%). The subsequent scale transfer of the C. laurentii UCD 68-201 lipid production process on RCW to a 3-L STR led to significantly improved biomass and total lipid productions (14.4 and 9.9gL-1, respectively) with the biodiesel yield amounting to 32.6%. Although the C. laurentii FAME profile was modified upon process transfer, it resembled that of the Jatropha oil, a well established feedstock for biodiesel production. In conclusion, C. laurentii UCD 68-201, for which there is very limited amount of available information, turned out to be a very promising candidate for biodiesel production and wide margins of process improvement might be envisaged.

The key role of meteorites in the formation of relevant prebiotic molecules in a formamide/water environment.

We show that carbonaceous chondrite meteorites actively and selectively catalyze the formation of relevant prebiotic molecules from formamide in aqueous media. Specific catalytic behaviours are observed, depending on the origin and composition of the chondrites and on the type of water present in the system (activity: thermal > seawater > pure). We report the one-pot synthesis of all the natural nucleobases, of aminoacids and of eight carboxylic acids (forming, from pyruvic acid to citric acid, a continuous series encompassing a large part of the extant Krebs cycle). These data shape a general prebiotic scenario consisting of carbonaceous meteorites acting as catalysts and of a volcanic-like environment providing heat, thermal waters and formamide. This scenario also applies to the other solar system locations that experienced rich delivery of carbonaceous materials, and whose physical-chemical conditions could have allowed chemical evolution.