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Aims: The work investigates the impact of grapes processing at the beginning of winemaking on the composition of microbiota during the oenological fermentations and on the composition of obtained wines. Methods and Results: The experiments were conducted in a biodynamic winery to exclude interference due to microbial starters. Three winemaking protocols, with different pre‐fermentative management of grapes, were followed by plate count and next generation sequencing on 16S for bacteria and internal transcribed spacer sequencing (ITS) for yeast. Chemical and sensory characterization of wine was performed. The grape processing influenced the evolution of microbiota (especially lactic and acetic acid bacteria) and the fermentation rate. The highest biodiversity was observed in the experiment carried out with whole grapes and carbonic maceration, with the presence of bacterial groups not usually found in winemaking (Bacteroidales, Clostridiales, Oscillospira). The different microbiotas influenced the organic acid profile of wines, the content of biogenic ammines and the perception of organoleptic descriptors linked to the vine cultivar (Syrah). Conclusions: Carbonic maceration impacts on the evolution of the microbiota and the wine features. The absence of addition of starters and sulphur dioxide would seem to be correlated with the high microbial biodiversity. Significance and Impact of the Study: Carbonic maceration is a traditional winemaking practice, today there are difficulties in its managing because the anaerobiosis stimulates spoilage micro‐organisms. The work elucidates the reasons of these difficulties and identified some microbial groups rarely associated with winemaking. The ratio of ethanol accumulation along with physical management of grapes and the supply of oxygen during the early stages of winemaking are powerful instruments of oenological variability, able to offer new possibilities to winemakers in order to defining the quality of red wines. [ABSTRACT FROM AUTHOR]

Abstract: Aims: Straw wine is a highly valuable oenological production, affected by relevant economical losses due to proliferation of spoilage micro‐organisms during drying grapes, after being harvested. In this work, ozone was evaluated as a tool to preserve grapes during drying, in terms of both qualitative and quantitative changes induced in the epiphytic microflora. In addition, the alteration exerted by ozone on grapes’ chemical composition was analysed. Methods and Results: Grapes from four vine varieties were treated with ozone produced by a cold plasma generator during the entire drying period (6 weeks). The microflora was quantified weekly by plate counts and characterized by 454‐pyrosequencing, and was compared with identical, untreated grape samples. At the end of drying, an extensive chemical characterization of the whole mass of grapes by FT‐IR and GC‐MS was performed. Ozone counteracted the growth of microflora by reducing the microbial population up to 3 log units. From the qualitative point of view, ozone reduced the incidence of spoilage micro‐organisms, such as Acetobacter and Botrytis cinerea, among the microbiota of grapes. Conclusions: The statistical analysis discriminates grape samples based on the residual epiphytic microflora at the end of drying and not on their chemical composition. These evidences confirmed that ozone did control spoilage micro‐organisms without altering the volatile profile of grapes. Chemical analysis revealed that untreated grapes are less suitable for winemaking owing to the deprivation of some valuable compounds during the microbial proliferation. Significance and Impact of the Study: Ozone is an emerging tool in winemaking, recently applied for controlling spoilage micro‐organisms in winery. The present study describes a new application of ozone as an effective and safe alternative to chemical preservatives which are actually involved in the control of microbial alterations of grapes. [ABSTRACT FROM AUTHOR]

To examine the influence of different types of exertion on immediate and delayed attention in 116 primary school children divided in three groups of exertion [cognitive exertion - CE (school curricular lesson), physical exertion - PE (traditional physical education lesson), mixed cognitive and physical exertion - CPE (coordinative physical education lesson)]. CPE was the combination of physical load due to the practice of physical exercises and of cognitive load requested to perform movement-based problem solving tasks requiring accurate timing, temporal estimations, temporal production, and spatial adjustments. Children's attentional capacity was tested before (pre) and after (at 0 min and at 50 min post) a CE, a PE, or a CPE lesson, using the d2-test of attention, and analyzed using a 3 × 3 × 2 mixed analysis of covariance with exertion type and time as within factors, gender as between factor, and baseline data as covariate. Effect sizes were calculated as partial eta squared (ƞ2). Results showed that participants' attentional performance was significantly affected by exertion type ( P < 0.0001), by time ( P < 0.0001) and by exertion type × time interactions ( P < 0.0001). The effect sizes ranged from medium (0.039) to large (0.437). Varying the type of exertion has different beneficial influences on the level of attention in school children. [ABSTRACT FROM AUTHOR]

The aims of this study were to estimate the difference in exercise intensity (METs), energy cost (EE) and gender diff erence between a typical salsa lesson (TSL), rueda de casino lesson (RCL), and salsa dancing at a night club (SDN). Subjects performed 1 pre-testing session and 3 testing conditions. During the pre-testing session height, weight and VO 2max were assessed. During the testing conditions all subjects performed 3 diff erent kinds of salsa dance. Heart rate was assessed during each dance condition. The exercise intensity of the 3 salsa dancing conditions was moderate ranging from 3.9 to 5.5 METs. A signifi cant diff erence between genders for HR peak (P = 0.01), max %HRR (P = 0.006) and mean EE (P = 0.02) were observed. Signifi cant gender × condition interactions for HR 2max (P = 0.03), mean %HRR (P = 0.02), mean METs (P = 0.02) and mean EE (P = 0.02) were found. In addition, a signifi cant main eff ect for each condition was found in all variables (P < 0.01). Our results showed that the exercise intensities of all 3 salsa dancing conditions were moderate. Findings showed some signifi cant diff erences in exercise intensity between males and females and within conditions. Salsa dancing could be useful in achieving a signifi cant training eff ect in people who have a low level of fi tness [ABSTRACT FROM AUTHOR]

Abstract: The microbial composition and its spatial distribution of Grana Trentino, a hard Parmesan-like cheese, was determined, from vat milk to cheese. After cutting along the vertical axis of the cheese wheels, three layers were sampled diagonally across the cheese: under the cheese rind, an intermediate section and the cheese core. After two different ripening periods (9 and 18 months), the cheese samples were analysed using traditional culture dependent and culture independent methods. Milk samples were dominated by mesophilic and psychrophilic bacterial counts. Thermophilic bacteria (Lactobacillus helveticus) were found in high amounts in cooked whey and natural whey starter cultures. After 9 months of ripening, lactic acid bacteria (LAB) counts were higher than those after 18 months. Furthermore, the LAB numbers in the cheese core was lower than those under the rind or in the intermediate section. The main LAB species isolated from milk (Lactococcus lactis, Pediococcus pentosaceus, Streptococcus uberis and Lactococcus garvieae) were not found in the corresponding cheeses. Some differences were observed in the species composition among the three cheese sections. Microbiota under the rind and in the intermediate section was similar and dominated by Lactobacillus paracasei and Lactobacillus rhamnosus. The core, after 18 months of ripening, was characterized by a total absence of LAB. In each sample, all LAB were genotypically grouped and the different biotypes were subjected to several technological tests indicating that some non-starter LAB (NSLAB) displayed technological features that are favorable for the production of Grana Trentino cheese. [Copyright &y& Elsevier]

Raw cow's milk and cheese samples collected from factories producing “Puzzone di Moena” located in the alpine province of Trento (northern Italy), were analyzed for their enterococcal content. Presumptive enterococci (Gram positive, catalase negative, coccal shaped and positive for growth on kanamycin aesculin azide) were isolated, verified for their growth in presence of 6.5% NaCl, 40% bile salts and esculin hydrolysis, and analyzed by a genetic polyphasic approach consisting of intergenic spacer (ITS)-polymerase chain reaction (PCR) between the 16S and 23S rRNA genes, denaturing gradient gel electrophoresis (DGGE) of the V3 region of the 16S rRNA gene and a sodA gene-based multiplex PCR. Results showed the presence of Enterococcus faecalis as the main species found in milk and cheese. Enterococcus casseliflavus and Enterococcus durans were also detected in raw milk. In order to exclude clonal relatedness, the isolates were subjected to randomly amplified polymorphic DNA-PCR (RAPD-PCR). Winter cheese was characterized by a higher biodiversity than summer production, both in terms of Enterococcus species and number of strains. No strain was found positive for vancomycin resistance as determined by vanA - and vanB -specific PCR. Two strains were shown to be characterized by an anti-Listeria behavior due to bacteriocin-like inhibitory substances (BLIS) production. BLIS producer strains persisted during the ripening of cheese. PRACTICAL APPLICATIONS Most of the traditional cheeses are made from raw milk, whose lactic acid bacterial population plays a major role during ripening. Enterococci from milk may survive the adverse hostile environment in cheese during ripening and be potentially transferred to consumers. Enterococcus strains isolated from milk involved in cheese production, and from ripened “Puzzone di Moena” cheeses, were shown not to host van genes, thus, they should not be involved in the process of antimicrobial resistance dissemination among intestinal microflora. Furthermore, it was found that bacteriocinogenic enterococci dominated over nonbacteriocin-like inhibitory substances producer Enterococcus strains during the cheese process, confirming previous studies showing that bacteriocin production may influence bacterial complexes. From an application standpoint, these results are basic in the selection of nonstarter lactic acid bacteria to be used as adjunct cultures in traditional cheese productions. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate sport-specific basketball skills before and after 8 months of integrated and non-integrated basketball practice of participants with intellectual disability; in relation to the competitive basketball level and the degree of intellectual disability., Design: Pre-test/training/post-test design., Methods: Forty-one adult male players with intellectual disability were randomly divided into 21 athletes playing in the Integrated Basketball group together with 10 athletes without intellectual disability, and 20 athletes playing in the Non-integrated Basketball group. All players were assessed through pre and post basketball skill tests for assessing four levels of ability of increasing difficulty (levels I, II, III, and IV), each one characterized by the fundamental skills of the basketball game: ball handling, passing, receiving, and shooting. The athlete's global score based on the total score of all levels of ability was calculated for each player., Results: Passing, receiving, shooting, ball handling, global, level I, and level II scores improved after the intervention independently by integrated basketball or non-integrated basketball. Post-pre (∆) scores in ball handling, receiving, passing, shooting, global, level I, and level II showed that the athletes in the Integrated Basketball group improved significantly more than athletes in the Non-integrated Basketball group. Ball handling, receiving, passing, shooting, global, level I, level II, and level III scores were negatively correlated with intellectual disability level., Conclusions: Athletes with intellectual disability who performed both integrated basketball and non-integrated basketball improved significantly their basketball skills after an 8-month training. However, the athletes training in the Integrated Basketball group obtained the best scores., Competing Interests: Declaration of interest statement The authors have no conflicts of interest to report., (Copyright © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The aim of this study was to develop health-oriented fermented salamis by replacing synthetic preservative (E 252) with dried Sumac (Rhus coriaria) fruit powder (DSFP). The salamis were produced at an industrial scale using meat from the "Suino Nero dei Nebrodi" breed, without adding starter cultures. The experimental design included four different salami productions: CTR, control production without nitrate salt and DSFP; CMC, commercial control production with nitrate salt but without DSFP; EXP1, experimental production without nitrate salt but with DSFP; and EXP2, experimental production with both nitrate salt and DSFP. Plate counts showed that DSFP did not inhibit the growth of lactic acid bacteria (LAB), coagulase-negative staphylococci, and yeasts, all of which reached approximately 7.0 log CFU/g in 45 d ripened salamis. Except for the CTR production, Escherichia coli levels decreased to undetectable amounts at 30 d of ripening. Culture-independent methods identified 16 taxonomic groups, with LAB being the predominant group across all trials, comprising 46.05-81.81 % of relative abundance (RA) in 45 d ripened salamis. Physicochemical analysis indicated that adding DSFP increased antioxidant activity by nearly 30 % and reduced primary lipid oxidation to levels comparable to those achieved with nitrate salt. The addition of DSFP in CMC, EXP1, and EXP2 salamis resulted in an approximate 11 % increase in total terpene aromatic profiles. Sensory evaluation indicated that the addition of DSFP did not impact overall acceptability (p > 0.05). Therefore, incorporating DSFP in fermented meat production offers a viable alternative to the use of synthetic preservatives., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Cow milk microbiota has received increased attention in recent years, not only because of its importance for human health but also because of its effect on the quality and technological properties of milk. Several studies, therefore, have investigated the effect of various production factors on the microbial composition of milk. However, most of the previous studies considered a limited number of animals from experimental or single farm, which could have biased the results. Therefore, this study aimed to understand the effect of different alpine production systems in Italy on the compositional and microbiological quality of milk, considering commercial herds with different feeding intensities and cattle breeds. The results obtained in this work indicated that the month and season of sampling (July for summer or February for winter) more than farm, breed, and cow diet exerted significant effects on cow milk parameters and microbiota. In particular, significant differences were observed for urea content in milk between sampling seasons. Differences in milk fat were mainly related to breed specific effects. From a microbiological point of view, statistically significant differences were found in presumptive lactic acid bacteria counts. Based on a culture-independent method, milk obtained in February harbored the highest number of Firmicutes (e.g., Lactobacillus) and the lowest number of Actinobacteria (e.g., Corynebacterium). Moreover, bacterial richness and diversity were higher in July during alpine pasture season indicating a significant effect of pasture feed on the growth of bacterial communities. The results of this study highlighted the effect of month or season mainly related to differences in feeding management (e.g., access to pasture during vegetation period, concentrates supplementation) on composition and microbiota in milk., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Sedentary behaviours in adults with intellectual disabilities (ID) negatively impact health-related determinants and increase the risk of physical dysfunction or chronic health issues. Inclusive sport practice can enhance social inclusion and fitness, benefiting overall person development and life quality. This study aimed to assess fitness level variation in athletes with and without ID before and after a 9-month inclusive (INC) and non-inclusive (N&#95;INC) basketball training. 38 athletes with ID and 14 athletes without ID belonged to INC group, 38 athletes with ID belonged to N&#95;INC group and 23 participants with ID belonged to the control group (CG). Before and after the intervention period, all participants performed anthropometric (body weight, body mass index, body fat percentage) and fitness tests (muscle strength and power, cardiovascular endurance, balance, flexibility, agility, movement speed, and coordination). ID and non-ID athletes of both INC and N&#95;INC groups significantly improved in most of the tests (weight, balance, flexibility, muscle strength, endurance, agility, movement speed and coordination) while CG group significantly worsened weight, muscle endurance, flexibility, agility, movement speed and coordination, after the intervention (p < 0.05). These findings showed that sport training improved fitness in athletes with ID, decreasing health risks. Moreover, involving in the same training group persons with and without disabilities, does not limit the beneficial effects that training induce on persons without ID and could be a valid way to promote both social inclusion and physical health of persons with ID., (© 2024. The Author(s).)

The aim of this study was to develop a novel and healthier fermented meat product by replacing pork fat with avocado pulp (AVP) during salami production. Experimental salamis were produced under laboratory conditions by substituting pork fat with AVP partially (10-AVP) and totally (20-AVP), while control salamis (CTR) remained AVP-free. The microbial composition of control and experimental salamis was assessed using a combined culture-dependent and -independent approach. Over a 20-days ripening period, lactic acid bacteria, coagulase-negative staphylococci, and yeasts dominated the microbial community, with approximate levels of 9.0, 7.0 and 6.0 log CFU/g, respectively. Illumina technology identified 26 taxonomic groups, with leuconostocs being the predominant group across all trials [constituting 31.26-59.12 % of relative abundance (RA)]. Gas Chromatography-Mass Spectrometry (GC-MS) analysis revealed changes in fatty acid composition and volatile organic compounds due to the substitution of pork fat with AVP. Specifically, monounsaturated fatty acids and terpene compounds increased, while saturated fatty acids and lipid oxidation products decreased. Although AVP influenced the sensory characteristics of the salamis, the highest overall satisfaction ratings were observed for the 10-AVP salamis. Consequently, substituting pork fat with AVP emerges as a viable strategy for producing healthier salamis and diversifying the meat product portfolio., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This work was carried out with the aim to reduce the transformation duration of Protected Designation of Origin (PDO) Pecorino Siciliano cheese. To this purpose, the cooking in hot water (experimental production, EXP) was compared to the traditional cheese cooking under whey permeate (control production, CTR). The microbiological composition of under rind (UR) and core (Co) section of CTR and EXP cheeses was determined by a combined culture-dependent and -independent approach. Total mesophilic microorganisms and lactic acid bacteria (LAB) present in raw ewes' milk (5.0 log CFU/mL) increased during cheese making and reached values of about 8.0 log CFU/g in both sections (UR and Co) of 5-month ripened cheeses of both productions (CTR and EXP) monitored. The identification of the viable LAB populations in ripened cheeses showed that Enterococcus, Lacticaseibacillus, Lactiplantibacillus, Levilactobacillus, Limosilactobacillus and Streptococcus dominated UR and Co sections of all cheeses. MiSeq Illumina analysis demonstrated that LAB populations (lactobacilli, lactococci and streptococci) dominated the bacterial community of cheeses at 95.63-98.41 % of relative abundance. The two different cooking operations did not influence the physicochemical characteristics of PDO Pecorino Siciliano cheeses. Sensory evaluation performed by artificial senses analysis and trained panelists confirmed that the modification of PDO Pecorino Siciliano cheese production protocol did not significantly affect product characteristics and overall acceptance. Thus, data of this work confirmed that cooking under hot water allowed to reduce transformation duration and safeguard typicality of PDO Pecorino Siciliano cheese., Competing Interests: Declaration of competing interest No conflict of interest exists., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The rind acts as a protective barrier for internally-bacterial ripened cheeses. Unlike surface-inoculated smear cheeses, centripetal maturation is not assumed to occur in these cheeses. This research was aimed to evaluate the microbial diversity of the wooden shelves used for the ripening of Protected Denomination of Origin (PDO) Pecorino di Filiano and Protected Geographical Indication (PGI) Canestrato di Moliterno cheeses. The microorganisms associated with the rind of these cheeses were also investigated. Both wooden shelf surfaces and cheese rinds were sampled by brushing method to collect their biofilms. Wooden shelves showed levels of total mesophilic microorganisms (TMM) between 5.6 and 7.2 log CFU/cm 2 , while cheese rinds between 6.1 and 7.8 log CFU/cm 2 . The major dairy pathogens (Salmonella spp., Listeria monocytogenes, Escherichia coli, and Staphylococcus aureus) were never detected, while mesophilic and thermophilic bacteria dominated the surfaces of all wooden shelves and cheese rinds. LAB community was represented by Enterococcus spp., Leuconostoc spp., and Marinilactibacillus spp. Among yeasts, Debaryomyces spp., Candida spp., were identified, while Aspergillus spp., and Penicillium spp., dominated the community of filamentous fungi. MiSeq Illumina analysis identified 15 phyla, 13 classes, 28 orders, 54 families, and 56 genera among bacteria. Staphylococcus spp. was identified from all wooden surfaces, with a maximum abundance of 71 %. Brevibacterium, Corynebacterium and halophilic bacteria were detected in almost all samples. Regarding fungi, wooden shelves mainly hosted Aspergillus, Penicillium and Debaryomyces hansenii, while cheese rinds especially Penicillium and D. hansenii. Alpha diversity confirmed a strict correlation between the microbiota of wooden shelves and that of cheese rinds for the majority of factories. This study confirmed that the wooden shelves used for cheese ripening are microbiologically active and represent safe systems. Furthermore, the results of this work clarified the transfer flow between wooden shelves and PDO Pecorino di Filiano and PGI Canestrato di Moliterno cheese surfaces: smear-active microorganisms are mainly transferred from wooden shelves to cheese rind, which potentially contribute to the development of the final organoleptic characteristics; meanwhile, cheeses transfer LAB that are potentially involved in defining the safety aspects of the shelves., Competing Interests: Declaration of competing interest No conflict of interest exists., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In this study, Caciocavallo Podolico Lucano (CPL) cheese was deeply characterized for its bacterial community, chemical composition and sensory aspects. The entire cheese making process (from milk collection to ripened cheese) was performed by strictly applying the traditional protocol for CPL production in four dairy factories (A-D) representative of the production area. The vat made of wood represents the main transformation tool for CPL cheese production and the biofilms hosted onto the internal surfaces of all vats analyzed in this study were dominated by lactic acid bacteria. Total mesophilic microorganisms present in bulk milk (4.7-5.0 log CFU/ml) increased consistently after contact with the wooden vat surfaces (5.4-6.4 log CFU/ml). The application of Illumina sequencing technology identified barely 18 taxonomic groups among processed samples; streptococci and lactobacilli constituted the major groups of the wooden vat biofilms [94.74-99.70 % of relative abundance (RA)], while lactobacilli dominated almost entirely (94.19-100 % of total RA) the bacterial community of ripened cheeses. Except coagulase positive staphylococci, undesirable bacteria were undetectable. Among chemical parameters, significant variations were registered for unsaturated, monounsaturated, polyunsaturated fatty acids and antioxidant properties (significantly lower for CPL cheeses produced in factory B). The cheeses from factories A, C and D were characterized by a higher lactic acid and persistence smell attributes than factory B. This work indicated that the strict application of CPL cheese making protocol harmonized the main microbiological, physicochemical and sensory parameters of the final cheeses produced in the four factories investigated., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Almond production generates large amounts of by-products rich in polyphenols. In this study, almond skin was explored as a valuable food ingredient in bread making. To this purpose, almond skin was used to produce functional products modifying a traditional sourdough bread recipe. The doughs were prepared replacing semolina with powdered almond skin (PAS) at 5 and 10 % (w/w). Sourdough inoculum was started with a mix of lactic acid bacteria (LAB) and propagated in semolina until reaching pH 3.7. The pH of PAS added breads was higher than that of control (CTR) breads before and after fermentation. Plate counts showed a similar evolution of LAB and total mesophilic microorganisms, but members of Enterobacteriaceae and coliform were detectable in PAS doughs. Illumina data clearly showed a dominance of lactobacilli in all trials, but PAS doughs displayed the presence of Bacillus. The final bread characteristics were influenced by PAS and its addition percentage; in particular, crust and crumb colour resulted darker, the alveolation decreased and, regarding sensory attributes, odour intensity increased, while bread odour diminished. In presence of PAS, bread emissions were characterized by lower percentages of alcohols and aromatic hydrocarbons and higher percentages of the other volatile compound classes, especially terpenoids like β-pinene, β-myrcene and limonene than CTR trial. After in vitro simulated digestion, the final release of phytochemicals from 10 % PAS bread was almost 100 %. Thus, PAS determined an increase of the antioxidant capacity of the breads. Phytochemicals released from digested PAS-fortified bread can provide antioxidant protection in a complex biological environment such as human intestinal-like cells. Besides the positive functional properties of PAS, this work also evidenced the hygienic issues of almond skin and, in order to avoid potential risks for the human health, highlighted the need to preserve its microbiological characteristics during storage for their reuse in bread production., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In this study, PDO Provola dei Nebrodi cheese was deeply characterized for its bacterial community and chemical composition. Four dairy factories (A-D) were monitored from milk to ripened cheese. Wooden vat biofilms were dominated by thermophilic rod LAB (4.6-6.5 log CFU/cm 2 ). Bulk milk showed consistent levels of total mesophilic microorganisms (TMM) (5.0-6.0 log CFU/mL) and, after curdling, a general increase was recorded. The identification of the dominant LAB in wooden vat biofilms and ripened cheeses showed that the majority of wooden vat LAB were lactococci and Streptococcus thermophilus, while cheese LAB mainly belonged to Lacticaseibacillus paracasei and Enterococcus. Illumina sequencing identified 22 taxonomic groups; streptococci, lactococci, lactobacilli and other LAB constituted the majority of the total relative abundance % of the wooden vat (69.01-97.58 %) and cheese (81.57-99.87 %) bacterial communities. Regarding chemical composition, the effect of dairy factories was significant only for protein content. Inside cheese color was lighter and yellower than surface. Differences in fatty acids regarded only myristic acid and total amount of monounsaturated fatty acids. The sensory evaluation indicated some differences among cheeses produced in the four dairies regarding color, homogeneity of structure, overall intensity, salty, spicy, and hardness. The integrated approach applied in this study showed that PDO Provola dei Nebrodi cheese characteristics are quite stable among the dairy factories analyzed and this has to be unavoidably imputed to the application of the same cheese making protocol among different dairies., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Introduction: In this study, we supplemented models of Caciotta-like cheese with blackcurrant ( Ribes nigrum ) and Cornelian cherry ( Cornus mas ), as they have a high content of polyphenols, known as phytochemicals associated with health benefits. We evaluated the microbial composition, organoleptic aspects, total phenolic content, and chemical composition of model cheeses enriched with blackcurrant and Cornelian cherry., Methods: Two different suppliers have been tested: a conventional and an organic one. Two different conditions of preparation (freeze-dried and not freeze-dried) were tested in two different amounts (0.3 and 0.6% dry weight w/v milk volume). Polyphenols were determined using Folin-Ciocalteu reaction and spectrometry; microbial community was determined with selective 24 media and plate counts; composition was determined using nuclear magnetic resonance spectrometry. Organoleptic tests with an untrained panel have been performed., Results: The enrichments with blackcurrant and Cornelian cherry increased the total polyphenol content in model cheeses, in particular, when blackcurrant and Cornelian cherry were from conventional farming. Blackcurrant-enriched cheeses showed higher counts of lactic acid bacteria, higher levels of organic acids, amino acids, gamma-aminobutyric acid, histamine, and lower amount of monosaccharides deriving from bacterial lactose fermentation in cheese, suggesting a positive effect of blackcurrant compounds on the growth and activity of lactic acid bacteria. The enrichments did not affect the acceptance of the cheese, neither by blackcurrant nor by Cornelian cherry incorporation, with the exception of the appearance., Discussion: Overall, we showed that cheeses enriched with blackcurrant or Cornelian cherry from conventional farming increased the bioactive potential of the dairy product without having an adverse effect on the microbial community, physiochemical properties, or organoleptic properties., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Andersen, Bosetti, Mancini, Solovyev, Nardin, Bontempo, Larcher and Franciosi.)

Bacteria have a fundamental role in determining the fitness of grapevine, the composition of grapes and the features of wines but at present, little information is available. In this work, the bacteria colonizing the different portions of grapevine (bark, leaves and grapes) were explored in the vineyards of the Alpine region of Trentino, considering the impact of different environmental and agronomical variables. The vineyards included in the work were selected based on their different geographical positions (altitude) and grapevine training systems in order to explore the whole variability of the grapevine ecosystem. Moreover, the surface amount of copper was measured on grapes and leaves during the vegetative growth. Bacterial analysis, performed using plate counts and Illumina MiSeq, revealed an increase in the concentration of grape bacteria proportional to the progress of the ripening stage. Conversely, the peak of bacterial concentration onto leaf and bark samples occurred in August, probably due to the more favourable environmental conditions. In bark samples, the bacterial microbiota reached the 7 log CFU/cm 2 , while 6 log UFC/g were measured in grape samples. A remarkable biodiversity was observed, with 13 phyla, 35 classes, 55 orders, 78 families and 95 genera of bacteria present. The presence of some taxa (Alphaproteobacteria, Desulfovibrionaceae, Clostriadiales, Oscillospira, Lachnospiraceae and Bacteroidales) was ubiquitous in all vineyards, but differences in terms of relative abundance were observed according to the vegetative stage, altitude of the vineyard and training system. Bacteria having oenological implication (Lactobacillus, Pediococcus and Oenococcus) were detected in grape samples collected in August, in low abundance. The data revealed a complex bacterial ecosystem inside the vineyard that, while maintaining common traits, evolves according to environmental and agronomical inputs. This study contributes to define the role of bacteria in the complex balance established in each vineyard between human actions and agricultural environment, known as terroir., (© 2022. The Author(s).)

The study of the complex relationships between milk metagenomics and milk composition and cheese-making efficiency as affected by indoor farming and summer highland grazing was the aim of the present work. The experimental design considered monthly sampling (over 5 mo) of the milk produced by 12 Brown Swiss cows divided into 2 groups: the first remained on a lowland indoor farm from June to October, and the second was moved to highland pastures in July and then returned to the lowland farm in September. The resulting 60 milk samples (2 kg each) were used to analyze milk composition, milk coagulation, curd firming, and syneresis processes, and to make individual model cheeses to measure cheese yields and nutrient recoveries in the cheese. After DNA extraction and Illumina Miseq sequencing, milk microbiota amplicons were also processed by means of an open-source pipeline called Quantitative Insights Into Microbial Ecology (Qiime2, version 2018.2; https://qiime2.org). Out of a total of 44 taxa analyzed, 13 bacterial taxa were considered important for the dairy industry (lactic acid bacteria, LAB, 5 taxa; and spoilage bacteria, 4) and for human (other probiotics, 2) and animal health (pathogenic bacteria, 2). The results revealed the transhumant group of cows transferred to summer highland pastures showed an increase in almost all the LAB taxa, bifidobacteria, and propionibacteria, and a reduction in spoilage taxa. All the metagenomic changes disappeared when the transhumant cows were moved back to the permanent indoor farm. The relationships between 17 microbial traits and 30 compositional and technological milk traits were investigated through analysis of correlation and latent explanatory factor analysis. Eight latent factors were identified, explaining 75.3% of the total variance, 2 of which were mainly based on microbial traits: pro-dairy bacteria (14% of total variance, improving during summer pasturing) and pathogenic bacteria (6.0% of total variance). Some bacterial traits contributed to other compositional-technological latent factors (gelation, udder health, and caseins)., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Sauerkraut is a traditionally fermented cabbage, and recent evidence suggests that it has beneficial properties for human health. In this work, a multi-disciplinary approach was employed to characterize the fermentation process and gut health-promoting properties of locally produced, organic sauerkraut from two distinct producers, SK1 and SK2. 16S rRNA metataxonomics showed that bacterial diversity gradually decreased as fermentation progressed. Differences in sauerkraut microbiota composition were observed between the two producers, especially at the start of fermentation. Lactic acid bacteria (LAB) dominated the microbiota after 35 days, with Lactiplantibacillus being the dominant genus in both sauerkraut products, together with Leuconostoc and Paucilactobacillus in SK1, and with Pediococcus, Levilactibacillus , and Leuconostoc in SK2. LAB reached between 7 and 8 Log CFU/mL brine at the end of fermentation (35 days), while pH lowering happened within the first week of fermentation. A total of 220 LAB strains, corresponding to 133 RAPD-PCR biotypes, were successfully isolated. Lactiplantibacillus plantarum and Lactiplantibacillus pentosus accounted for 67% of all SK1 isolates, and Lactiplantibacillus plantarum/paraplantarum and Leuconostoc mesenteroides represented 72% of all the isolates from SK2. 1 H-NMR analysis revealed significant changes in microbial metabolite profiles during the fermentation process, with lactic and acetic acids, as well as amino acids, amines, and uracil, being the dominant metabolites quantified. Sauerkraut brine did not affect trans-epithelial electrical resistance through a Caco-2 cell monolayer as a measure of gut barrier function. However, significant modulation of inflammatory response after LPS stimulation was observed in PBMCs-Caco-2 co-culture. Sauerkraut brine supported a robust inflammatory response to endotoxin, by increasing TNF-α and IL-6 production while also stimulating the anti-inflammatory IL-10, therefore suggesting positive resolution of inflammation after 24 h and supporting the potential of sauerkraut brine to regulate intestinal immune function., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gaudioso, Weil, Marzorati, Solovyev, Bontempo, Franciosi, Bertoldi, Pedrolli, Tuohy and Fava.)

This research aimed to analyze variations in chemical properties, microbiological characteristics and generated volatile organic compounds (VOCs) profile during sourdough fermentation. Sourdoughs were collected from different cities in Turkey at two different times and lactic acid bacteria (LAB) in the samples were identified with culture-independent and culture-dependent molecular methods. According to culture-dependent methodology, thirteen LAB species were identified. Lactobacillus spp. were identified as the major group according to MiSeq Illumina analysis. Technological potential of commonly isolated LAB species was evaluated. Due to high frequency of isolation, Fructilactobacillus sanfranciscensis and Lactiplantibacillus plantarum strains were better investigated for their technological traits useful in sourdough production. Experimental sourdoughs were produced with mono- and dual-culture of the selected strains and chemical properties and microbiological characteristics, as well as VOCs profile of the sourdoughs, were subjected to multivariate analysis which showed the relevance of added starter, in terms of acidification and VOCs profile., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

During a survey for isolating sourdough lactic acid bacteria (LAB), 20 dough samples produced at the bakery level (BL) or home-made (HM) were collected. An enzyme-based colorimetric method revealed a total biogenic amines (BAs) concentration in the range 41.4-251.8 ppm for six (three BL and three HM) sourdoughs characterised by unpleasant odours. Eight BAs generally investigated in foods were identified and quantified from these six samples by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Only one HM sample contained almost all analysed BAs. Tryptamine was exclusively detected in HM sourdoughs (0.71-24.1 ppm). Putrescine, tryptamine, spermidine, and spermine were the only BAs detected in BL sourdoughs. MiSeq Illumina analysis was applied to study the total bacterial community of sourdoughs. LAB accounted from 67.89 to 92.17% of total bacterial diversity, and Levilactobacillus brevis was identified in all six sourdoughs. Leuconostoc , Pediococcus , and Weissella were also dominant. Plate counts detected neither the presence of Pseudomonas nor members of the Enterobacteriaceae family, and LAB levels were, on average, barely 5.89 Log CFU/g for BL, and 7.33 Log CFU/g for HM sourdoughs. Data suggested that the microorganisms mainly imputable of BAs formation in sourdough are members of the LAB community.

Bacteria can play different roles affecting flavors and food characteristics. Few studies have described the bacterial microbiota of butter. In the present paper, next-generation sequencing was used to determine bacterial diversity, together with aromatic characteristics, in raw cow milk butter processed by traditional fermentation, in fourteen small farms called "Malga", located in the Trentino province (Alpine region, North-East of Italy). The physicochemical and aromatic characterization of traditional mountain butter (TMB) showed a low moisture level depending on the Malga producing the butter. Counts of lactic acid bacteria, Staphylococci , and coliforms, as well as diacetyl/acetoin concentrations exhibited changes according to the geographical origin of Malga and the residual humidity of butter. MiSeq Illumina data analysis revealed that the relative abundance of Lactococcus was higher in TMB samples with the highest values of acetoin (acetoin higher than 10 mg/kg). The traditional mountain butter bacterial community was characterized by a "core dominance" of psychrotrophic genera, mainly Acinetobacter and Pseudomonas , but according to ANCOM analysis, a complex bacterial population emerged and specific bacterial genera were able to characterize the TMB bacteria community, with their high abundance, based on the Malga producing the butter.

The main goal of this research was to characterize the bacterial diversity of the wooden boards used for aging traditional Sicilian cheeses and to evaluate whether pathogenic bacteria are associated with these surfaces. Eighteen cheese dairy factories producing three traditional cheese typologies (PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano) were selected within the region of Sicily. The wooden shelf surfaces were sampled by a destructive method to detach wood splinters as well as by a nondestructive brushing to collect microbial cells. Scanning electron microscopy showed the presence of almost continuous bacterial formations on the majority of the shelves analyzed. Yeasts and fungal hyphae were also visualized, indicating the complexity of the plank communities. The amplicon library of the 16S rRNA gene V3-V4 region was paired-end sequenced using the Illumina MiSeq system, allowing the identification of 14 phyla, 32 classes, 52 orders, 93 families, and 137 genera. Staphylococcus equorum was identified from all wooden surfaces, with a maximum abundance of 64.75%. Among cheese-surface-ripening bacteria, Brevibacterium and Corynebacterium were detected in almost all samples. Several halophilic ( Halomonas , Tetragenococcus halophilus, Chromohalobacter, Salimicrobium , Marinococcus , Salegentibacter , Haererehalobacter , Marinobacter , and Idiomarinaceae ) and moderately halophilic ( Salinicoccus , Psychrobacter , and Salinisphaera ) bacteria were frequently identified. Lactic acid bacteria (LAB) were present at low percentages in the genera Leuconostoc , Lactococcus , Lactobacillus , Pediococcus , and Streptococcus. The levels of viable microorganisms on the wooden shelves ranged between 2.4 and 7.8 log CFU/cm 2 . In some cases, LAB were counted at very high levels (8.2 log CFU/cm 2 ). Members of the Enterobacteriaceae family were detected in a viable state for only six samples. Coagulase-positive staphylococci, Salmonella spp., and Listeria monocytogenes were not detected. Seventy-five strains belonged to the genera Leuconostoc , Lactococcus , Pediococcus , Enterococcus , Lactobacillus , and Weissella . IMPORTANCE This study provides evidence for the lack of pathogenic bacteria on the wooden shelves used to ripen internal bacterially ripened semihard and hard cheeses produced in Sicily. These three cheeses are not inoculated on their surfaces, and surface ripening is not considered to occur or, at least, does not occur at the same extent as surface-inoculated smear cheeses. Several bacterial groups identified from the wooden shelves are typically associated with smear cheeses, strongly suggesting that PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano cheese rind contributes to their final organoleptic profiles.

Purpose: In this paper, the authors present insights and findings drawn from the authors' experiences of containing a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak in a large prison in northern Italy.Within penitentiaries, close-quarter living is ripe terrain for outbreaks of disease among detainees and staff. If left unchecked, these outbreaks can easily spill over the prison walls to threaten the general public. Moreover, these risks are heightened by preexisting environmental conditions, especially overcrowding. It is thus paramount to establish effective protocols for prevention, early detection and outbreak management. The purpose of this article is to document a strategy that been at least partially successful in reducing the damage that could potentially be caused by a sustained SARS-CoV-2 outbreak within a correctional facility., Design/methodology/approach: The authors conducted a retrospective analysis on patients' and health-care workers' medical records to obtain demographic and clinical information. Descriptive data analysis was then carried out., Findings: In total, the authors tested 453 people with oropharyngeal swabs from March 15, 2020, to June 30, 2020. Of these people, 58 were positive and 395 were negative, with a prevalence of 12.8%.Of the 453 patients, 60 were health workers: 24 tested positive for SARS-CoV2 ribonucleic acid (RNA); 18 developed symptoms; and three needed hospitalization.Among patients in detention, 34 resulted positive for SARS-CoV-2 RNA. Two were hospitalized and later died. Both had severe preexisting conditions; they were aged 76 and 59 years old, respectively., Originality/value: In this study, the authors describe the design and effective implementation of prevention and containment measures against SARS-CoV-2 within the walls of a correctional facility. The authors describe how they rapidly created clean confinement sections to isolate cases in an environment designed for security at the expense of virus containment and how educational efforts have played a vital role in their strategy., (© Emerald Publishing Limited.)

This work was carried out to elaborate selenium (Se) bio-enriched fermented Mediterranean fruit juices. To this purpose, pomegranate and table red grape juices were added with sodium selenite (Na 2 SeO 3 ) and fermented by Levilactobacillus brevis CRL 2051 and Fructobacillus tropaeoli CRL 2034 individually or combined. To better evaluate the effect of selenite addition and starter strain inoculums on the total bacterial community of the fruit juices, fermentation trials were performed with raw and pasteurized fruit juices. No statistical significant differences were observed for total mesophilic microorganisms (TMM) and rod-shaped lactic acid bacteria (LAB) levels among raw and pasteurized juices inoculated with the starter strains, while significant differences between those juices with and without selenite were registered. LAB cocci, Pseudomonadaceae and yeasts were detected only for the raw juice preparations. The dominance of L. brevis CRL 2051 and F. tropaeoli CRL 2034 was confirmed by randomly amplified polymorphic DNA (RAPD)-PCR analysis. After fermentation, pH dropped for all inoculated trials and control raw juices. The soluble solid content (SSC) levels of the raw juices were higher than the corresponding pasteurized trials. The thermal treatment affected consistently yellowness of grape juice trials and redness of pomegranate juices. No microbial Se accumulation was registered for pomegranate juices, while F. tropaeoli CRL 2034 accumulated the highest amount of Se (65.5 μg/L) in the grape juice. For this reason, only trials carried out with raw grape juices were investigated by metagenomics analysis by Illumina MiSeq technology. Non-inoculated grape juices were massively fermented by acetic acid bacteria while Fructobacillus and Lactobacillus (previous genus name of Levilactobacillus) represented the highest operational taxonomy units (OTUs) relative abundance % of the trials inoculated with the starter strains as confirmed by this technique., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: The symptoms and complications related to chronic liver disease (CLD) have been shown to affect patient well-being. Currently there is limited research data on how CLD severity may affect both health-related quality of life (HRQOL) and the development of depressive symptoms in CLD patients. Moreover, the ongoing advances in CLD treatment, and its effect on HRQOL, highlight the need for further studies. Therefore, the aim of the present study was to evaluate if the CLD severity may affect the HRQOL and the development of depressive symptoms., Methods: A cross-sectional study was conducted. Patients with CLDs were identified at their regular visits to the outpatient clinic of the Sant'Orsola-Malpighi Hospital in Bologna, between September 2016 and July 2017. HRQOL was measured with Short Form 12 (SF-12) and Nottingham Health Profile (NHP) questionnaires; depressive symptoms were measured with Beck Depression Inventory-II (BDI). CLD severity was measured using the MELD score and the sample was stratified into five classes according to it. Group comparisons were conducted using the Kruskal-Wallis test., Results: Two hundred and fifty-four patients were included. Mean age was 62.84 years (SD 11.75) and 57.9% were male. Most participants were affected by compensated cirrhosis (140.2%) and chronic hepatitis (40.2%), with a disease duration ≥ 5 years (69.3%). Regarding the MELD score, 67.7% of patients belonged to Class I, 29.9% to Class II, and 2.4% to Class III. There were not patients belonging to the Classes IV and V. No statistically significant differences were found in all SF-12 and NHP domains between the MELD classes, except for CLD impact on sexual life and holidays (p = 0.037 and p = 0.032, respectively). A prevalence rate of 26% of depressive symptoms was reported, no statistically significant differences were found in BDI-II total scores between the three MELD classes., Conclusions: All domains of HRQOL and depression were altered in CLDs patients, nevertheless CLD severity was not confirmed as an affecting factor for HRQOL., (© 2021. The Author(s).)

This study focused on the microbial and bacteriophages identification and characterization in cheese-production facilities that use natural whey starter (NWS) cultures for Trentingrana production. Bacterial and phage screening was carried out on cooked not acidified whey and NWS samples isolated from six dairy factories, for 4 consecutive days in four different months. By means of a combined approach, using plate counts, bacterial isolation, and metataxonomic analysis Lactobacillus helveticus was found occurring as the dominant species in NWS cultures and Levilactobacillus brevis as codominant in the cheese factories where the temperature of NWS production was mainly lower than 40°C, suggesting that the variability in the parameters of the NWS culture preparation could differently modulate the bacterial species in NWS cultures. Using turbidity test approach on 303 bacterial isolates from the NWS cultures, 120 distinct phages were identified. L. helveticus phage contamination of NWS cultures was revealed in most of the analyzed samples, but despite the great recovery of bacteriophage contamination cases, the microbial quality of NWS cultures was high. Our results support the presence of natural bacteriophage resistance mechanisms in L. helveticus . The use of NWS cultures probably creates an ideal environment for the proliferation of different L. helveticus strains balanced with their phages without a clear dominance. It is evident, from this study, that the presence of a high biodiversity of NWS bacterial strains is relevant to avoid phages dominance in NWS cultures and consequently to keep a good acidification ability., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Mancini, Rodriguez, Zago, Cologna, Goss, Carafa, Tuohy, Merz and Franciosi.)

Competing Interests: Competing interests: None declared.

Phyto-thermotherapy is a treatment consisting in immersing oneself in baths of self-heating alpine grass, to benefit of the heat and rich aromatic components released by the process. The aim of this study was to characterize the bacterial and fungal diversity of three phyto-thermal baths (PTB) performed in three different months, and to compare the data with the profile of the volatile organic compounds (VOCs) of the process. All the data collected showed that PTBs were structured in two stages: the first three days were characterised by an exponential rise of the temperature, a fast bacterial development, higher microbial diversity and higher concentrations of plant aliphatic hydrocarbons. The second stage was characterised by a stable high temperature, shrinkage of the microbial diversity with a predominance of few bacterial and fungi species and higher concentrations of volatiles of microbial origin. Erwinia was the dominant microbial species during the first stage and probably responsible of the self-heating process. In conclusion, PTBs has shown both similarities with common self-heating processes and important peculiarities such as the absence of pathogenic bacteria and the dominance of plant terpenoids with health characteristics among the VOCs confirming the evidence of beneficial effects in particular in the first three days.

In the present study, two groups of cows from a permanent lowland farm (PF) were divided during summer and reared in the PF or in a temporary alpine farm (ALP), respectively. Microbiological analyses were performed with the objective to investigate the microbial evolution of milk before, during, and after summer transhumance comparing, in particular, the two groups of cows to determine whether the alpine pasture could directly influence the milk microbiota. A significant increase of all microbial groups was registered in milk samples collected in the ALP. Interestingly, many strains belonging to species with well reported technological and probiotic activities were isolated from Alpine milk (20% Lactococcus lactis subsp. lactis/cremoris, 18% Lactobacillus paracasei, 14% Bifidobacterium crudilactis and 18% Propionibacterium sp.), whereas only 16% of strains isolated from the permanent farm milk belonged to the species Lactococcus lactis subsp. lactis/cremoris, 6% to Lactobacillus paracasei, 2% to Bifidobacterium crudilactis and 5% to Propionibacterium sp. The MiSeq Illumina data showed that Alpine milk presented a significant reduction of Pseudomonas and an increase of Lactococcus, Bifidobacterium and Lactobacillus genera. These data confirmed the practice of Alpine pasture as one of the main drivers affecting the milk microbiota. All the microbial changes disappeared when cows were delivered back from Alpine pasture to the indoor farm., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Fermented chickpea liquid is used as a leavening agent in chickpea bread production. In the present study, traditional chickpea liquid starter and dough samples were collected from bakeries in Turkey and microbiologically investigated. Culture-independent analysis for microbiota diversity, performed by MiSeq Illumina, identified Clostridium perfringens as major group in all samples, while Weissella spp. Dominated LAB community. A culture-dependent methodology was applied and 141 isolates were confirmed to be members of the LAB group based on 16s rRNA gene sequence analysis. In particular, 11 different LAB species were identified confirming the high frequency of isolation of weissellas, since Weissella confusa and Weissella cibaria constituted 47.8 and 12.4%, respectively, of total LAB isolated. The other species were Enterococcus faecium, Enterococcus lactis, Lactobacillus brevis, Lactobacillus plantarum, Leuconostoc mesenteroides, Leuconostoc mesenteroides subsp. Dextranium, Pediococcus acidilactici, Pediococcus pentosaceus and Streptococcus lutetiensis. Due to high frequency of isolation, W. confusa strains were investigated at technological level and W. confusa RL1139 was used as mono-culture starter in the experimental chickpea sourdough production. Chemical and microbiological properties, as well as volatile organic compounds (VOCs) of the chickpea liquid starters and doughs were subjected to a multivariate analysis. Control and W. confusa inoculated chickpea liquid starter and dough samples were close to each other in terms of some characteristics related to chemical, microbiological and VOCs profile, but the inoculated sourdough showed a higher generation of certain VOCs, like butanoic acid (81.52%) and ethyl acetate (8.15%) than control sourdough. This is important in order to maintain typical characteristics of the traditional chickpea dough, but at the same time improving the aroma profile. This work demonstrated that W. confusa RL1139 can be applied at large scale production level without compromising the typical characteristics of the final product., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present study was carried out to produce Protected Denomination of Origin (PDO) Pecorino Siciliano cheese with a multi-species lactic acid bacteria (LAB) culture, composed of starter and non-starter strains in order to reduce the microbiological variability of the products derived without LAB inoculums. To this end, cheese samples produced in six factories located in five provinces (Agrigento, Catania, Enna, Palermo and Trapani) of Sicily, and previously characterised for physicochemical, microbiological and sensory aspects, have been investigated in this work for bacterial microbiome, fatty acid (FA) composition as well as volatile organic compound (VOC) profiles. Analysis of the cheese microbiomes indicated that streptococci (30.62-77.18% relative abundance) and lactobacilli (on average 25.90% relative abundance) dominated the bacterial communities of control cheeses, produced without exogenous inoculums, whereas the cheeses produced with the selected multi-strain culture saw the dominance of lactococci (in the range 6.49-14.92% relative abundance), streptococci and lactobacilli. After the addition of the selected mixed culture, Shannon index increased in all cheeses, but only the cheeses produced with the selected LAB mixed culture in the factory 2 showed Gini-Simpson diversity index (0.79) closer to the reference value (0.94) for a perfect even community. FA composition, mainly represented by saturated FA (on average 69.60% and 69.39% in control cheeses and experimental cheeses, respectively), was not affected by adding LAB culture. The presence of polyunsaturated FA ranged between 7.93 and 8.03% of FA. VOC profiles were different only for the content of butanoic acid, registered for the experimental cheeses at higher concentrations (on average 662.54 mg/kg) than control cheeses (barely 11.96 mg/kg). This study validated addition of the ad hoc starter/non-starter culture for PDO Pecorino cheese production., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In the present work the microbial dynamics in wheat kernels were evaluated over time. The main aim of this research was to study the resistance of lactic acid bacteria (LAB) and yeasts associated to unprocessed cereals used for bread making during long term conservation. To this purpose four Triticum durum Desf. genotypes including two modern varieties (Claudio and Simeto) and two Sicilian wheat landraces (Russello and Timilia) were analysed by a combined culture-independent and -dependent microbiological approach after one, two or three years from cultivation and threshing. DNA based MiSeq Illumina technology was applied to reveal the entire bacterial composition of all semolina samples. The samples showed a different distribution of bacterial taxa per variety and time of storage. The groups mostly represented were Stenotrophomonas, Pseudomonas, Erwinia, Delftia and Sphingomonas genera, Enterobacteriaceae and Oxalobacteriaceae families, and Actinobacteria phylum. Among LAB, only Enterococcus genus was detected barely in a single sample (Simeto stored for one year) by the next generation sequencing, indicating that LAB remained unassigned or their abundances were below 0.1% or their DNAs were rendered inaccessible. Plate counts showed consistent differences in relation to genotypes and duration of storage, with the highest levels found for total mesophilic microorganisms detected up 6.8 Log CFU/g. Colonies of presumptive sourdough LAB were detected only in a few samples. Cocci constituted the major group of LAB in almost all samples. Following the enrichment procedure, almost all samples were characterised by the presence of acidifying microorganisms. All isolates collected before and after enrichment represented 28 different strains belonging to 10 species of Enterococcus, Lactobacillus and Pediococcus genera. The most resistant species during aging were Enterococcus faecium, Enterococcus durans, Lactobacillus brevis, Lactobacillus pentosus and Lactobacillus paracasei demonstrating that lactobacilli and enterococci are able to overcome the stressing conditions represented by cereal storage better than other LAB genera commonly found associated to cereals after harvest. Yeast community included mostly species with no interest in bread making., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The present work was carried out to evaluate the microbiological and physicochemical composition of salamis produced with the meat of beef, horse, wild boar and pork. Salami productions occurred under controlled laboratory conditions to exclude butchery environmental contaminations, without the addition of nitrate and nitrite. All trials were monitored during the ripening (13 °C and 90% relative humidity) extended until 45 d. The evolution of physicochemical parameters showed that beef and pork salamis were characterized by a higher content of branched chain fatty acids (FA) and rumenic acid than horse and wild boar salamis, whereas the last two productions showed higher values of secondary lipid oxidation. Plate counts showed that lactic acid bacteria (LAB), yeasts and coagulase-negative staphylococci (CNS) populations dominated the microbial community of all productions with Lactobacillus and Staphylococcus as most frequently isolated bacteria. The microbial diversity evaluated by MiSeq Illumina showed the presence of members of Gammaproteobacteria phylum, Moraxellaceae family, Acinetobacter, Pseudomonas, Carnobacterium and Enterococcus in all salamis. This study showed the natural evolution of indigenous fermented meat starter cultures and confirmed a higher suitability of horse and beef meat for nitrate/nitrite free salami production due to their hygienic quality at 30 d., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

An 80-year-old woman presented with a several-year history of progressive hair loss and scalp pruritus. No other rashes or muscle weakness were noted on examination. Scalp biopsy showed interface dermatitis, dense perivascular and periadnexal lymphocytic infiltrate, mucin and scarring alopecia. Laboratory analysis did not show evidence of myositis. The patient was started on hydroxychloroquine for possible cutaneous lupus erythematosus. On follow-up, she presented with a new violaceous rash on the superior eyelids and a well-defined oval patch on the mid-hard palate suspicious for dermatomyositis. Myositis-specific autoantibodies revealed presence of anti-transcriptional intermediary factor-1γ (anti-TIF1γ) in the serum. Anti-TIF1γ autoantibody-positive dermatomyositis is a newly recognised subtype of dermatomyositis that is highly associated with amyopathic disease and has an increased risk of malignancy, making prompt diagnosis crucial. This case highlights the utility of a thorough oral exam in patients suspected to have connective tissue disease as the distinctive ovoid palatal patch is nearly pathognomonic for anti-TIF1γ dermatomyositis., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Pistachio powder was added to flour or semolina to evaluate its contribution to increase the amount of lysine in bread. Bread production was carried out by sourdough technology using a selected 3-species (Lactobacillus sanfranciscensis/Leuconostoc citreum/Weissella cibaria) lactic acid bacterial (LAB) starter culture. All sourdoughs were subjected to a long-time fermentation (21 h) and showed levels of LAB around 10 9  CFU/g, indicating the suitability of pistachio powder for lactic fermentation. Yeasts were also detected, in particular in semolina trials. MiSeq Illumina technology was applied to investigate the bacterial composition of sourdoughs evidencing a different distribution of LAB species among the trials with Lactobacillus as major LAB group in almost all sourdoughs. Physicochemical parameters were comparable among the trials. After baking, pistachio powder was found not to influence the height of the breads, but pistachio breads were more firm than control breads. Color of the breads, void fraction and cell density, were influenced by pistachio powder. The amount of lysine increased consistently thanks to pistachio supplementation which also determined a higher presence of o-xylene, p-cymene and limonene and the appearance of α-pinene and 1-octen-3-ol in breads. Sensory tests showed the best appreciation scores for the breads produced with flour and pistachio powder., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In the present research, bacterial diversity was studied during a 6-month feeding trial utilizing zebrafish (Danio rerio) fed Hermetia illucens reared on different substrates with an emphasis on fish gut bacterial diversity. A polyphasic approach based on viable counting, PCR-DGGE and metagenomic 16S rRNA gene amplicon target sequencing was applied. Two different H. illucens groups were reared on coffee by-products (C) or a mixture of vegetables (S). Viable counts showed a wide variability based on substrate. PCR-DGGE and Illumina sequencing allowed the major and minor bacterial taxa to be detected. Both samples of larvae and their frass reared on the S substrate showed the highest richness and evenness of bacterial communities, whereas zebrafish (ZHC) fed H. illucens reared on substrate C and zebrafish (ZHS) fed H. illucens reared on substrate S had the lowest bacterial richness and evenness. A stimulating effect of bioactive compounds from coffee by-products on the occurrence of Lactobacillaceae and Leuconostoccaceae in H. illucens reared on substrate C has been hypothesized. Zebrafish gut samples originating from the two feeding trials showed complex microbial patterns in which Actinobacteria and Alteromonadales were always detected, irrespective of the diet used. Enterobacteriaceae in fish guts were more abundant in ZHS than in ZHC, thus suggesting an influence of the bioactive compounds (chlorogenic and caffeic acids) in the substrate on Enterobacteriaceae in fish guts. ZHC showed a higher abundance of Clostridia than did ZHS, which was likely explained by stimulating activity on the bacteria in this class by the bioactive compounds contained in H. illucens reared on substrate C. An influence of the microbiota of H. illucens or insect-derived bioactive compounds on the gut microbiota of zebrafish has been suggested. The presence of bacteria consistently associated with zebrafish guts has been found irrespective of the diet, thus attesting to the likely stability of the core fish microbiota., Competing Interests: The authors have declared that no competing interests exist.

This work was carried out to pursue a double objective: to improve the hygienic safety of cheeses produced from raw ewes' milk; and to produce a new typology of raw ewes' milk through the application of "Grana" technology for which the name "Gran Ovino" was chosen. With this in mind, raw milk from an individual farm was transformed under controlled conditions at a dairy pilot plant. The production technology included the partial skimming of the evening and morning milk mixture by cream surfacing and the addition of a natural whey starter cultures (NWSC) prepared with four selected Streptococcus thermophilus strains (PON6, PON244, PON261 e PON413). Ten microbial groups were investigated by plate counts from raw milk until ripened cheeses. Lactic acid bacteria (LAB) were in the range 10 4 -10 5  CFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 10 6  CFU/g after curd cooking. A rapid pH drop (to 6.05) was registered after almost 3 h from NWSC addition. The levels of members of the Enterobacteriaceae family were at about 10 3  CFU/ml in raw milk and decreased after curd cooking to 1 log cycle. A similar behavior was shown by the other undesired microbial groups and a complete disappearance of staphylococci was registered. The microbiological counts of 9-month ripened cheeses showed the dominance of LAB and undetectable levels of the undesired bacteria. MiSeq Illumina was applied to better investigate the bacterial composition of ripened cheeses and this technique evidenced that the majority of OTUs belonged to Lactobacillus and Streptococcus genera. The final cheeses were characterized by 67.65% dry matter of which 41.85% of fats and 47.02% of proteins. The main cheese fatty acids were palmitic, oleic and myristic acids and the saturated fatty acids/unsaturated fatty acids ratio was 2.17. Forty-one volatile compounds, including acids, esters, ketones, alcohols, aldehydes, phenols and one terpene were emitted from the cheese. Sensory evaluation showed a general appreciation for the new cheese product by judges., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Summer transhumance to alpine pastures (ALP) is widespread in dairy systems of alpine regions. This study aimed to investigate the effects of transhumance of Brown Swiss cows to ALP on the yield, composition, and coagulation properties of milk (MCP), and on cheese yield (CY). The study involved 12 multiparous cows kept at a mountain lowland permanent farm (PF), which were divided into two equal groups: One remained at the PF, the other was moved to the ALP (1860 m above sea level) from July to September. Every month (June to October), daily milk yield (MY) and body condition score (BCS) were recorded, and individual milk samples ( n = 60, 2000 mL each) were collected to assess milk composition, MCP, and CY. Compared with PF, ALP cows had a reduced MY and BCS, which was maintained on return to the PF, greater fat and lower protein contents of milk. Neither MCP nor CY were affected by summer transhumance. In conclusion, summer transhumance did not affect the cheese making efficiency of milk but depressed MY and consequently daily cheese yield, which was nearly 2 kg/d lower for the ALP than the PF cows and was only partially recovered after returning to the PF in autumn.

The cheese-derived strains Streptococcus thermophilus 84C isolated from Nostrano cheese, and Lactobacillus brevis DSM 32386 isolated from Traditional Mountain Malga cheese have been previously reported as γ-aminobutyric acid (GABA)-producers in vitro . In the present study, the ability of these strains to produce GABA was studied in experimental raw milk cheeses, with the aim to investigate the effect of the culture and the ripening time on the GABA concentration. The cultures used consisted on S. thermophilus 84C alone (84C) or in combination with L. brevis DSM 32386 (84C-DSM). The control culture was a commercial S. thermophilus strain, which was tested alone (CTRL) or in combination with the L. brevis DSM 32386 (CTRL-DSM). The pH evolution, microbiological counts, MiSeq Illumina and UHPLC-HQOMS analysis on milk and cheese samples were performed after 2, 9, and 20 days ripening. During the whole ripening, the pH was always under 5.5 in all batches. The concentration of GABA increased during ripening, with the highest content in 84C after 9 days ripening (84 ± 37 mg/kg), in 84C-DSM and CTRL-DSM after 20 days ripening (91 ± 28 and 88 ± 24 mg/kg, respectively). The data obtained support the hypothesis that S. thermophilus 84C and L. brevis DSM 32386 could be exploited as functional cultures, improving the in situ bio-synthesis of GABA during cheese ripening.

Lactococcus lactis subsp. lactis 68, Streptococcus thermophilus 93 and Lactobacillus rhamnosus BT68, previously isolated from Traditional Mountain (TM) cheese, were tested for the production of four experimental mountain cheeses, with the aim to assess their effectiveness in leading the TM-cheese-making process. Lactococcus lactis subsp. lactis 68 and Streptococcus thermophilus 93 were used as starter cultures, whereas Lactobacillus rhamnosus BT68 was used as non-starter culture. Three control (CTRL) cheeses were manufactured without adding any starter, according to the traditional cheese-making process; nine, cheeses were produced inoculating the vat milk with the starters (ST), starter and low concentration of non-starter culture (STLC), starter and high concentration of non-starter culture (STHC). Samples of vat milk, cheese after 24 h and 7 months ripening were processed for microbiological counts. Mesophilic cocci were dominant in all 24 h-cheese samples, while a dominance of both cocci and lactobacilli was observed after 7 months ripening. The total genomic DNA was extracted, and a fragment of V1-V3 region was amplified and pyrosequenced. Lactococci and streptococci were the most abundant species, and Lc. lactis ssp. lactis 68 affected the proliferation of milk-resident Lc. lactis ssp. cremoris, during the early fermentation. Lb. rhamnosus BT68 showed to be responsible in reducing the abundance of other Lactobacillus species. Moreover, it likely competed against Sc. thermophilus 93 for the same energetic sources, when added in concentration higher than 5 × 10 3  CFU/mL milk. The sensorial and fatty acid (FA) composition analysis were performed on cheese samples at the end of ripening, demonstrating that the inoculated cheeses had better sensorial characteristics (aspect, smell, taste, texture) than CTRL cheeses, and that Lb. rhamnosus BT68 at high concentration is related to the increase of short chain fatty acids and conjugated linoleic acid in cheese after 7 months ripening., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Phytothermotherapy ("grass baths") is a traditional phytotherapy for rheumatism consisting of taking baths in hot fermenting grass. Scientific studies have demonstrated its efficiency in treating several rheumatic diseases. However the efficiency and repeatability of the therapy is dependent on the wild fermentations, determining sometimes the appearance of unpleasant conditions leading to the early abandonment of the therapy. The metabolism undergoing in the grass baths is unknown and there is not an established method to evaluate and predict grass baths quality. The aim of this study is to establish a simple VOCs profiling method able to evaluate the grass baths, predicting their evolution, through the identification of marker volatiles related to the best conditions and/or the spoilage. After replicating in real scale the traditional grass baths, the volatile profiles were measured using passive diffusion samplers injected in a thermal desorption-comprehensive GC × GC-TOF-MS. The high dimensionality of the data coupled with the limited number of time points, required a rigorous method development for the analysis of the data, achieved through the development of a novel R package for variable selection in GC × GC data matrices. The further application of a fuzzy clustering approach demonstrated to be a useful tool dealing with short time series, allowing to discard un-trending volatiles and giving a clear snapshot of the main trends in the data. A broad coverage of the volatolome was provided, thus suitable to describe the main metabolic changes ongoing in the grass baths. Coupling this data with the temperature and pH, and comparing it to the data from similar processes, like silage and compost, we demonstrated that the established method can be helpful to evaluate short time series, allowing us to obtain a list of volatiles as candidate markers for the quality of the grass baths. The established method gave a list of markers applicable to real scale grass baths to predict spoilage; furthermore it provides a list of volatiles where to search for candidate markers with reported health-related effects and can be used to generate hypothesis on the mechanisms of action of the treatment., (Copyright © 2018. Published by Elsevier B.V.)