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Depleted gas reservoirs are a valuable option for underground hydrogen storage (UHS). However, different classes of microorganisms, which are capable of using free H2 as a reducing agent for their metabolism, inhabit deep underground formations and can potentially affect the storage. This study integrates metagenomics based on Illumina‐NGS sequencing of bacterial and archaeal 16S rRNA and dsrB and mcrA functional genes to unveil the composition and the variability of indigenous microbial populations of four Italian depleted reservoirs. The obtained mcrA sequences allow us to implement the existing taxonomic database for mcrA gene sequences with newly classified sequences obtained from the Italian gas reservoirs. Moreover, the KEGG and COG predictive functional annotation was used to highlight the metabolic pathways potentially associated with hydrogenotrophic metabolisms. The analyses revealed the specificity of each reservoir microbial community, and taxonomic and functional data highlighted the presence of an enriched number of taxa, whose activity depends on both reservoir hydrochemical composition and nutrient availability, of potential relevance in the context of UHS. This study is the very first to address the profiling of the microbial population and allowed us to perform a preliminary assessment of UHS feasibility in Italy. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the shelf-life of pre-packaged sheep's arrosticini produced in a factory located in northern Italy. Samples were divided into two series and packaged in modified atmosphere with specific gas mixtures: conventional (C: 35% O2/15% CO2/50% N2) and experimental (E: 30% CO2/70% N2). All the samples were stored at 4°C for 10 days and subjected, after 5, 8 and 10 days (t5, t8, t10), to triplicate microbiological and chemical- physical (pH, total volatile basic nitrogen, thiobarbituric acid reactive substances) analyses. At the same time, the colorimetric analysis and a sensorial evaluation were carried out (pack tightness, presence of color or odor alterations), assigning a discrete score from 0 to 5. The samples showed initial total bacterial counts close to 5 Log CFU/g: these values gradually increased during storage, exceeding the threshold of 8 Log CFU/g in the C series; lower values were detected in the E series. A similar trend was highlighted for Enterobacteriaceae, with initial loads of about 3 Log CFU/g and an increase until t10, reaching values higher than 6 Log CFU/g in the C series and close to 5 Log CFU/g in the E series (P=0.002). E. coli also showed a similar trend, although with values approximately 1 Log lower than Enterobacteriaceae. Pseudomonas spp. showed initial counts close to 4.5 Log CFU/g, with a different increase in the C series (6.5 Log CFU/g at t10) and the E series (4.95 Log CFU/g) (P= 0.006). The higher growth in the C series was also observed for lactic acid bacteria, with an increase from 3 to 5 Log CFU/g (3.8 Log CFU/g in the E series P=0.016). The other microbiological parameters showed very low counts and, in most cases undetectable counts (<2 Log CFU/g) for the entire period considered. The initial values obtained from the measurement of the colorimetric indices were within the norm for this type of product: however, starting from t5, lower values of red index and lightness in the E series were measured, with an evident greying of meat surface. The results of the sensory evaluation indicated that the product showed optimal sensorial characteristics up to 8 days of shelf-life in the C series, while the use of an oxygenfree atmosphere, despite having a moderate inhibiting effect on the microbial populations, has led to an early modification of the product (5 days of storage), due to the presence of superficial greyish areas. The microbiological characteristics of arrosticini strictly depend on the hygienic conditions of slaughtering and production; even in optimal situations, the product is particularly perishable, and requires careful management of storage temperatures and times, to maintain its quality characteristics. [ABSTRACT FROM AUTHOR]

Background: Tuber aestivum Vittad., known as black summer truffle, represents high-value food especially used as garnishment in nouvelle cuisine. The aim of this study was to investigate on the viable microbial populations associated with T. aestivum ascomata collected in different sites of Sicily and one locality of Umbria (Italy). Methods: The ripe ascomata of black summer truffles were collected from Central Italy. Cell densities of spoilage bacteria, fecal indicators, potential pathogens, yeasts, and molds were analyzed. Statistical analysis was conducted with XLSTAT software. Results: The microbiological counts of truffles ranged between 6.00 and 9.63 log Colony Forming Unit (CFU)/g for total mesophilic count and between 6.18 and 8.55 log CFU/g for total psychrotrophic count; pseudomonads were in the range 6.98-9.28 log CFU/g. Listeria spp. and coagulase-positive streptococci detected in no samples. Coagulase-negative streptococci were found in some samples with 2.11-4.76 log CFU/g levels. Yeasts and filamentous fungi were detected at consistent levels of 3.60-7.81 log CFU/g. Significant differences (p<0.01) were found between samples and also for all microbial groups. Conclusion: This study evidenced that the common brushing procedure applied for preparation of truffles is not sufficient to eliminate microbial risks for consumers. The application of an efficient decontamination treatment is strongly suggested before consumption of fresh truffles. [ABSTRACT FROM AUTHOR]

The production of wines with peculiar analytical and sensorial profiles, together with the microbiological control of the winemaking process, has always been one of the main objectives of the wine industry. In this perspective, the use of oenological starters containing non-Saccharomyces yeasts can represent a valid tool for achieving these objectives. Here we present the results of seven pilot scale fermentations, each of which was inoculated with a different non-Saccharomyces yeast strain and after three days with a commercial Saccharomyces cerevisiae starter. The fermentations were carried out in double on 70 L of Sangiovese grape must, the most widely planted red grape variety in Italy and particularly in Tuscany, where it is utilized for the production of more than 80% of red wines. Fermentations were monitored by assessing both the development of the microbial population and the consumption of sugars at the different sampling times. The impact of the different starters was assessed after stabilization through the evaluation of the standard analytical composition of the resulting wines, also taking into account polysaccharides and volatile compounds. Moreover, quantitative descriptive sensory analyses were carried out. Compared to the control wines obtained by inoculating the S. cerevisiae starter strain, those inoculated with non-Saccharomyces/Saccharomyces mixed starters presented a significant differentiation in the chemical-analytical composition. Moreover, sensory analysis revealed differences among wines mainly for intensity of color, astringency, and dryness mouthfeel perception. [ABSTRACT FROM AUTHOR]

[Display omitted] • Fermented liver sausages collected from twenty artisan producers were studied. • The bacterial biota displayed the highest frequency (80%) of Latilactobacillus sakei. • Debaryomyces hansenii , Staphylococcus xylosus , Staphylococcus equorum were found. • Main volatiles included terpenoids, aldehydes, ketones, alcohols, esters, and acids. • Hepatitis E virus assessed via real-time-RT-(q)PCR assay was never detected. The aim of the present study was to obtain information on the occurrence of bacteria and eumycetes in ready-to-eat fermented liver sausages manufactured by 20 artisan producers located in the Marche Region (Italy). To this end, culture-dependent analyses and metataxonomic sequencing were carried out. Physico-chemical parameters and volatilome of the fermented liver sausages were also studied. Finally, the presence of hepatitis E virus (HEV) was also assessed via real-time-RT-(q)PCR assays. Active microbial populations mainly represented by lactic acid bacteria, enterococci, coagulase-negative cocci, and eumycetes were detected. Enterobacteriaceae, Pseudomonadaceae, and sulfite-reducing anaerobes were not detected in most of the samples. Latilactobacillus sakei dominated in all the analyzed samples, reaching abundances up to 80%. Staphylococcus xylosus and Staphylococcus equorum were also detected. Among minority bacterial taxa, Weissella spp., Leuconostoc spp., Macrococcus caseolyticus , Brochothrix thermosphacta, Staphylococcus succinus , Lactobacillus coryniformis , Lactiplantibacillus plantarum , Lactococcus garviae, Psychrobacter spp. , and Carnobacterium viridans were detected. The mycobiota was mainly composed by Debaryomyces hansenii that was present in all samples at the highest frequency. Among minority fungal taxa, Aspergillus spp., Penicillium spp., Kurtzmaniella zeylanoides , Candida spp., Yamadazyma spp., Scopulariopsis spp., Yarrowia spp., and Starmerella spp. were detected. Interestingly, associations between some taxa and some physico-chemical parameters were also discovered. The absence of HEV in all the samples attested a high level of safety. Finally, most of the VOCs detected in the analyzed fermented liver sausages belonged to six classes as: terpenoids, aldehydes, ketones, alcohols, esters, and acids. Nitrogen compounds, sulfur compounds, phenols, hydrocarbons, lactones, furans, and aromatic hydrocarbons were also identified. Several significant relationships were observed between mycobiota and VOCs. [ABSTRACT FROM AUTHOR]

Ocean acidification, resulting from rising atmospheric carbon dioxide concentrations, is a pervasive stressor that can affect many marine organisms and their symbionts. Studies which examine the host physiology and microbial communities have shown a variety of responses to the ocean acidification process. Recently, several studies were conducted based on field experiments, which take place in natural CO vents, exposing the host to natural environmental conditions of varying pH. This study examines the sea anemone Anemonia viridis which is found naturally along the pH gradient in Ischia, Italy, with an aim to characterize whether exposure to pH impacts the holobiont. The physiological parameters of A. viridis ( Symbiodinium density, protein, and chlorophyll a+ c concentration) and its microbial community were monitored. Although reduction in pH was seen to have had an impact on composition and diversity of associated microbial communities, no significant changes were observed in A. viridis physiology, and no microbial stress indicators (i.e., pathogens, antibacterial activity, etc.) were detected. In light of these results, it appears that elevated CO does not have a negative influence on A. viridis that live naturally in the site. This suggests that natural long-term exposure and dynamic diverse microbial communities may contribute to the acclimation process of the host in a changing pH environment. [ABSTRACT FROM AUTHOR]

Abstract: In this study we examined the spatial and temporal dynamics of planktonic bacterial assemblages at four different sites in the Lagoon of Venice. Samples were collected in January, April, July and October 2005 and several parameters (temperature, salinity, dissolved oxygen, chlorophyll a, dissolved macronutrients, dissolved and particulate organic carbon, viral and heterotrophic nanoplanktonic abundances) were determined in order to highlight the most important factors which are implied in shaping such assemblages. Furthermore we tested the relationship between similar assemblages and the patterns of activities (prokaryotic carbon production and several hydrolytic activities) that they perform in order to establish if, in this highly variable environment, similar assemblages behave in analogous ways. Results indicate that seasonality act as the main forcing on the communities. Moreover, we found a mismatch between community structure and patterns of activity possibly as a consequence of the heterogeneity of the lagoon which can affect in turn the assemblages'' metabolic requirements (and thus their responses). [Copyright &y& Elsevier]

Abstract: In this study we investigated the microbiota of Gorgonzola rinds and maturing shelf swabs collected in 5 different maturing cellars in the Northwest part of Italy, in association with the detection and characterization of Listeria monocytogenes. Culture-dependent and -independent methods were performed in order to profile the main microbial populations present on the rinds and in the maturing shelves and species-specific PCR and Pulsed Field Gel Electrophoresis (PFGE) were used to identify and type L. monocytogenes isolates. The microflora was predominated by lactic acid bacteria and coagulase negative cocci, while enterococci and yeasts were very variable between the samples. Arthrobacter sp., Carnobacterium sp., Staphylococcus sp. and Brevibacterium linens, as bacteria, and Debaryomyces hansenii, as yeast, were detected by Denaturing Gradient Gel Electrophoresis (DGGE). Cluster analysis of the DGGE profiles clearly highlighted a cellar-specific microflora. L. monocytogenes was isolated in 11.1% of the rinds and 29.4% of the swabs and the molecular characterization of the isolates suggests a route of contamination from the maturing shelves to the rinds. No correlation was found between DGGE profiles and presence or absence of L. monocytogenes. [Copyright &y& Elsevier]

The changes and features of some olive cultivars during natural fermentation in brine were examined. The cultivars were typical of Sicily: "Nocellara messinese", "Nocellara etnea", "Moresca", "Ogliarola" and "Tonda iblea". The carpological data revealed their good quality. Chemical characteristics such as reducing sugars, titratable acidity, total phenols and dry matter were also investigated. The physical and chemical changes of olives and brines throughout the process time were monitored. Sodium chloride concentration was gradually increased from 5% (w/v) to 6% after 60-75 days and 7% after 90. The olives were put in brine acidified up to pH = 4. The salt concentration and pH value were necessary to favor lactic fermentation. An uniform trend of pH was measured for all the olive cultivars: a stable pH under 4.5 value was measured. The course of titratable acidity measured in fermentation brines was in agreement with the pH trend. The concentration of total phenols in flesh olives through the brine storage and fermentation is reported and the reduction of total phenols during the brine fermentation was fast in "Tonda iblea", "Nocellara etnea" and "Moresca" olives. To measure the degree of oleuropein disappearance, the course of hydroxytyrosol in brine was monitored. Throughout the brining period, the growth of microbial populations was revealed on selective media: lactic acid bacteria, mesophilic microorganisms, yeasts and moulds. [ABSTRACT FROM AUTHOR]

Waters from an extensive sulfide-rich aquifer emerge in the Frasassi cave system, where they mix with oxygen-rich percolating water and cave air over a large surface area. The actively forming cave complex hosts a microbial community, including conspicuous white biofilms coating surfaces in cave streams, that is isolated from surface sources of C and N. Two distinct biofilm morphologies were observed in the streams over a 4-year period. Bacterial 16S rDNA libraries were constructed from samples of each biofilm type collected from Grotta Sulfurea in 2002. β-, γ-, δ-, and ϵ-proteobacteria in sulfur-cycling clades accounted for ≥75% of clones in both biofilms. Sulfate-reducing and sulfur-disproportionating δ-proteobacterial sequences in the clone libraries were abundant and diverse (34% of phylotypes). Biofilm samples of both types were later collected at the same location and at an additional sample site in Ramo Sulfureo and examined, using fluorescence in situ hybridization (FISH). The biomass of all six stream biofilms was dominated by filamentous γ-proteobacteria with Beggiatoa-like and/or Thiothrix-like cells containing abundant sulfur inclusions. The biomass of ϵ-proteobacteria detected using FISH was consistently small, ranging from 0 to less than 15% of the total biomass. Our results suggest that S cycling within the stream biofilms is an important feature of the cave biogeochemistry. Such cycling represents positive biological feedback to sulfuric acid speleogenesis and related processes that create subsurface porosity in carbonate rocks. [ABSTRACT FROM AUTHOR]

The relationship between fluid geochemistry and microbial communities was investigated for shallow (< 25 m) submarine hydrothermal vents in the Aeolian Islands (Southern Italy). Thermal waters, gases, and deposits of white filamentous material were collected from 11 sites. The geochemical analyses showed a magmatic component was present in all sampled fluids. The total microbial abundances, evaluated as direct counts of picoplanktonic cells (ranging from 0.2 to 2 mu m in diameter), were between 1.55 107 and 4.18 108 cells per liter. Picophytoplankton (total autofluorescent cells) ranged from 9.6 105 to 7.88 106 cells per liter. Yellow-orange autofluorescent prokaryotes belonging to the cyanobacteria were more abundant than red autofluorescent eukaryotic cells. Chemolithoautotrophic, sulfur-oxidizing, rod-shaped Bacteria were isolated from venting water samples and identified as Thiobacillus -like. Microscopic examination of the white mat deposits showed the presence of filamentous microorganisms. [ABSTRACT FROM AUTHOR]

The standardisation of raw milk cheeses without losing their typicality is an ambitious goal. In the present study, the performance of two Lactococcus lactis autochthonous starter cultures (AS1 and AS2) was evaluated for the production of Valle d'Aosta (VdA, Italy) Fromadzo PDO, in terms of competition and persistence capacity (by rep-PCR strain characterisation), influence on indigenous microbial population (by metataxonomic approach) and volatilome development (by GC–MS and sensory analysis). The best performance was shown by AS2 starter mixture, which was able to colonise Fromadzo cheeses at the expense of indigenous lactic microbiota, and reached mean frequency values of 87% and 70% in 30 day-ripened products from valley floor and alpine pasture milk, respectively. Moreover, both kinds of cheeses, obtained using AS2, achieved the best scores in terms of aroma and taste, the pasture cheese also achieving the best score for texture. [ABSTRACT FROM AUTHOR]

Composting is a complex process in which various micro-organisms, mainly fungi and bacteria, are involved. The process depends on a large number of factors (biological, chemical, and physical) among which microbial populations play a fundamental role. The high temperatures that occur during the composting process indicate the presence of thermotolerant and thermophilic micro-organisms that are key for the optimization of the process. However, the same micro-organisms can be harmful (allergenic, pathogenic) for workers that handle large quantities of material in the plant, and for end users, for example, in the indoor environment (e.g., pots in houses and offices). Accurate knowledge of thermotolerant and thermophilic organisms present during the composting stages is required to find key organisms to improve the process and estimate potential health risks. The objective of the present work was to study thermotolerant and thermophilic mycobiota at different time points of compost maturation. Fungi were isolated at four temperatures (25, 37, 45, and 50 °C) from compost samples collected at five different steps during a 21-day compost-maturation period in an active composting plant in Liguria (northwestern Italy). The samples were subsequently plated on three different media. Our results showed a high presence of fungi with an order of magnitude ranging from 1 × 104 to 3 × 105 colony-forming units (CFU) g−1. The isolated strains, identified by means of specific molecular tools (ITS, beta-tubulin, calmodulin, elongation factor 1-alpha, and LSU sequencing), belonged to 45 different species. Several thermophilic species belonging to genera Thermoascus and Thermomyces were detected, which could be key during composting. Moreover, the presence of several potentially harmful fungal species, such as Aspergillus fumigatus, A. terreus, and Scedosporium apiospermum, were found during the whole process, including the final product. Results highlighted the importance of surveying the mycobiota involved in the composting process in order to: (i) find solutions to improve efficiency and (ii) reduce health risks. [ABSTRACT FROM AUTHOR]