Piacentinu Ennese PDO cheese as reservoir of promising probiotic bacteria

Piacentinu Ennese is a protected designation of origin (PDO) cheese produced in the surrounding area of Enna (Sicily, Italy), using raw ewe’s milk without the addition of any starter cultures. In the present study, the Lactobacillus population of Piacentinu Ennese PDO cheese was in vitro screened in order to select promising probiotic strains to be further used in humans. One hundred and sixty-nine lactic acid bacteria (LAB) were isolated from 90 days ripened cheeses and identified by Rep-PCR genomic fingerprinting, using the (GTG)5-primer, and by MALDI-TOF MS. One hundred and thirteen (113) isolates belonging to QPS-list species were characterized for both safety and functional properties. All tested isolates were considered safe because none showed either gelatinase, DNase, mucinase, or hemolytic activity. Tolerance to lysozyme, bile salts, and acidic conditions, along with ability to survive under simulated gastrointestinal digestion, were observed. In addition, based on antimicrobial activity against pathogens, cell surface characteristics, Caco-2 adhesion abilities, and anti-inflammatory potential, it was possible to confirm the strain-dependent functional aptitude, suggesting that Piacentinu Ennese PDO cheese may be considered a precious source of probiotic candidates

Profiling of phenol content and microbial community dynamics during pâté olive cake fermentation

In this study, different microbial strains, as single and mixed-cultures, were used to ferment the pâté olive cake (POC), a by-product of olive oil processing. In particular, strains belonging to Candida boidinii, Wickerhamomyces anomalus and Lactiplantibacillus plantarum were used. The fermentation was carried out on diluted (3:2) POC without and with glucose (2% w/v) addition. Furthermore, phenolic compounds were monitored during fermentation in POC added with glucose differetly inoculated and the microbial community, at the end of fermentation, was evaluated by Next Generation Sequencing (NGS) techniques. Data highlighted that inoculated samples showed an hydroxytyrosol content higher than the un-inoculated controls. In particular, during fermentation the sample inoculated with C. boidinii, both in single and in mixed culture together with L. plantarum, increased the hydroxytyrosol content by 275 and 261 mg/L, respectively, after 8 days, to reach the highest content at the end of fermentation. Metagenomic analysis revealed a low abundance of 16S ribosomal RNA genes and fungal ITS in all samples at any sampling times. Furthermore, at the end of fermentation, all samples exhibited a different bacterial community with a decrease in acetic acid bacteria and an increase in Lactobacillaceae biodiversity. Finally, no effect was detected in any samples on fungal metagenomic profile, where Dipodascus geotrichum was found dominant both at initial and final fermentation. In conclusion, the present study confirmed that selected cultures can drive the fermentation and have an impact on the phenolic profile.This study was conducted within a Ph.D. research programme in Biotecnologie (XXXV cycle) by Paola Foti who received a grant ‘Dottorato innovativo con caratterizzazione industriale, PON RI 2014–2020’, titled ‘Olive oil by-products as a new functional food and source of nutritional food ingredients’ from the Department of Agriculture, Food and Environment (Scientific Tutors: C.C. Cinzia Caggia; Flora V. Romeo and Cinzia L. Randazzo). Authors thank the Azienda Olearia Consoli Pasquale & F.lli s.n.c (Adrano, CT), partner of the doctoral programme, to kindly supply the POC. The present study was partially supported by a regional funding Progetto di investimento 144511020025. P.O. FESR SICILIA 2014/2020, 2019–2021, Azione 1.1.3 - Sostegno alla valorizzazione economica dell’innovazione attraverso la sperimentazione e l’adozione di soluzioni innovative nei processi, nei prodotti e nelle formule organizzative, nonch ́e attraverso il finanziamento dell’industrializzazione dei risultati della ricerca: VERIFICO, Project number: 07TP1039000074info:eu-repo/semantics/publishedVersio

Platelet Antiaggregating Activity and Chemical Constituents of Salvia x Jamensis J. Compton

A phytochemical study has been carried out on the surface exudate of Salvia x jamensis, which showed a significant platelet antiaggregating activity. The known compounds isopimaric acid (2), 14-α-hydroxy-isopimaric acid (3), 3β-hydroxy-isopimaric acid (4), 7,8β-dihydrosalviacoccin (5), betulinic acid (6), and ursolic acid (7) were isolated together with the new diterpene 1. The structure of 1 was determined as 15,16-epoxy-cleroda-3-en-7α,10β-dihydroxy-12,17;19,18-diolide on the basis of spectroscopic data analysis. Among all tested compounds, 2 showed a significant concentration-dependent antiaggregating activity when ADP (3 μM) was used as agonist on rat platelets. Conversely, 1 increased ADP–induced platelet aggregation

The Wide Range of Antibiotic Resistance and Variability of Genotypic Profiles in Escherichia coli from Domestic Animals in Eastern Sicily

serious public health threat. Escherichia coli, a usual host of intestinal microbiota, is recognized also as etiological agent of numerous infections widespread in both humans and animals. The colibacillosis is one of the most reported zoonoses worldwide, typically treated with antibiotics in the primary stages. This strategy has promoted the onset of antibiotic-resistant serotypes of E. coli, reducing the effectiveness of therapeutic treatments and contributing to antibiotic resistance spread. The current study focused on biodiversity, pathogenicity, and antibiotic resistance profile of 104 E. coli strains isolated from domestic animals in Eastern Sicily. The strains were isolated from sick animals and carcasses of six different animal species and screened for resistance against 16 antibiotic molecules, as recommended by WHO and OIE. The antibiotic resistance patterns highlighted that all strains were multi-resistant, showing resistance to at least three antibiotic classes. The highest incidence of resistance was observed against amoxicillin (100%), tylosin (97%), sulfamethoxazole (98%), and erythromycin (92%), while the lowest for colistin (8%). The pathotype characterization identified two EPEC strains and the study of genetic linkage (PFGE) showed a wide variety of profiles. The current study emphasized the wide range of multidrug resistance and genotyping profiles in E. coli isolated in Easter Sicily

Chemical Profiling of Astragalus membranaceus Roots (Fish.) Bunge Herbal Preparation and Evaluation of Its Bioactivity

Astragalus membranaceus (Fish.) Bunge is a perennial herb distributed in the northern part of China, and its roots, namely, Hang qi, are included as a natural ingredient in dietary supplement formulations commonly used to treat different disorders such as respiratory infections, diabetes, and heart failure. The availability of a simple method for the determination of the quality of Astragalus herbal preparations could be a challenging issue for commercial purposes. In this study, a liquid chromatography–mass spectrometry (LC–MS)/MS based approach was used to characterize specialized metabolite recovery of 3 commercial hydroalcoholic extracts of A. membranaceus (AMG1, AMG2, AMG3) in addition to a hydroalcoholic extract of A. membranaceus root (AST). The hypoglycemic effect, cholinesterase inhibition, and antioxidant activities were also evaluated. Thirty-one compounds, of which 19 polyphenols and 12 saponins, were identified. The extracts were also quantified by using a sensitive and selective Q-Trap system for their content in flavonoids and astragalosides, selecting astragaloside I and IV as chemical markers. From our results, AMG3 preparation (Axtragyl) was the most abundant in terms of both specialized classes of metabolites, showing a fingerprint similar to that of AST. Interestingly, tested enzyme inhibition ability of flavonoids, daidzein (11) and formononetin (19), reported a higher α-glucosidase inhibition in comparison with that of acarbose used as positive control. The in silico study clarified the interactions among the molecules and the importance of having a free hydroxy group. Moreover, Axtragyl was able to exert protective effects in Caco-2 cells treated with hydrogen peroxide, confirming its ability as a potential protective agent in intestinal injury

Effect of olive by-products feed supplementation on physicochemical and microbiological profile of Provola cheese

IntroductionWith the purpose to evaluate the effects of dietary olive cake, a source of bioactive phenolic compounds, as feed supplementation of lactating dairy cows on fatty acid composition, volatile organic compounds, and microbiological profiles of Provola cheese, we performed a two-arm study where control and experimental administered cows derived dairy have been compared.MethodsOur panel of analyses include metabolomics, physicochemical detected variables, culture dependent and independent analyses, and a stringent statistical approach aimful at disclosing only statistically significant results.Results and discussionLooking at the physicochemical variable’s profiles, a higher content of unsaturated fatty acids, polyunsaturated fatty acid, and conjugated linoleic acids as well of proteins were observed in experimental cheese samples, indicating the beneficial effect of dietary supplementation. Furthermore, based on volatilome composition, a clear cluster separation between control and experimental cheeses was obtained, mainly related to terpenes degradation, able of influencing their aroma and taste. Microbiological results showed a decrease of some spoilage related microbial groups in experimental cheeses, probably due to the inhibitory effect exerted by polyphenols compounds, that contrarily did not affect the core taxa of all cheese samples. This paper confirmed the promising utilization of olive by-product in farming practices to obtain more sustainable and safe dairy food products with lower environmental impact, mainly in Sicily and Mediterranean area, where waste disposal poses serious environmental and economic problems

Influence of olive cake dietary supplementation on fecal microbiota of dairy cows

Olive by-products represent a valuable low-price feed supplement for animal nutrition. In the present study, the effect of the dietary destoned olive cake supplementation, on both composition and dynamics of the fecal bacterial biota of cow, was assessed by Illumina MiSeq analysis of the 16S rRNA gene. In addition, metabolic pathways were predicted by using the PICRUSt2 bioinformatic tool. Eighteen lactating cows, according to the body condition score, the days from calving, and the daily milk production were homogeneously allocated into two groups, control or experimental, and subjected to different dietary treatments. In detail, the experimental diet contained, along with the components of the control one, 8% of destoned olive cake. Metagenomics data revealed significant differences in abundance rather than in richness between the two groups. Results showed that Bacteroidota and Firmicutes were identified as the dominant phyla, accounting for over 90% of the total bacterial population. The Desulfobacterota phylum, able to reduce sulfur compounds, was detected only in fecal samples of cows allocated to the experimental diet whereas the Elusimicrobia phylum, a common endosymbiont or ectosymbiont of various flagellated protists, was detected only in cows subjected to the control diet. In addition, both Oscillospiraceae and Ruminococcaceae families were mainly found in the experimental group whereas fecal samples of control cows showed the presence of Rikenellaceae and Bacteroidaceae families, usually associated with the high roughage or low concentrate diet. Based on the PICRUSt2 bioinformatic tool, pathways related to carbohydrate, fatty acid, lipid, and amino acids biosynthesis were mainly up regulated in the experimental group. On the contrary, in the control group, the metabolic pathways detected with the highest occurrence were associated with amino acids biosynthesis and degradation, aromatic compounds degradation, nucleosides and nucleotides biosynthesis. Hence, the present study confirms that the destoned olive cake is a valuable feed supplement able to modulate the fecal microbiota of cows. Further studies will be conducted in order to deepen the inter-relationships between the GIT microbiota and the host

Effect of Molasses and Dried Orange Pulp as Sheep Dietary Supplementation on Physico-Chemical, Microbiological and Fatty Acid Profile of Comisana Ewe's Milk and Cheese

The use of agro-industrial by-products for ruminant feed represents both an economical and environmental convenient way for reducing waste discharge and waste management costs for food industries. Large amounts of waste from citrus processing industries are available in Sicily, Italy. In the present study, the effect of dried citrus pulp as sheep dietary supplementation was evaluated on physico-chemical, microbiological and fatty acid composition of resulting milk and cheese. Pelleted feed integrated with molasses and blond orange pulp, replacing cane molasses, beet pulp and part of the maize and sunflower in ration, were administrated to ewes as an experimental treatment The experiment involved sixty Comisana breed sheep divided into two groups and two feeding trials (experimental and control). Ewe's milk and cheese samples were collected from January to April and analyzed for physico-chemical, microbiological and fatty acid profile composition. Results suggested that both the experimental milk and cheese were different from the controls. In particular, an increase of experimental milk yield and fat content were registered whilst the cheese samples exhibited a significant decrease of pH values and an increase in fat and protein contents. In addition, an increase of conjugated linoleic acids as well as of the oxidative stability were observed indicating the beneficial effect of dietary supplementation. Furthermore, among the main microbial groups, the experimental and control samples, no differences were detected. However, with the exception of streptococci, which was found higher in experimental cheeses, and staphylococci, which was significantly reduced by experimental feed. Moreover, the application of culture-independent methods highlighted the dominance of Lactobacillus rhamnosus/casei group in the experimental cheese, suggesting a driving role of the dietary supplementation in the cheese microbiota composition. The present study demonstrated that the inclusion of citrus by-products in the diet of small dairy ruminants is a promising feeding, which could positively affect milk composition and cheese manufacture

Towards the post-antibiotic era: The role of antibiotic resistant enterococci in different sources

L esplosione del fenomeno dell antibiotico resistenza (AR) si configura come un fenomeno globale. Sebbene i batteri AR sia stati per lungo tempo esclusivamente associati ad ambienti ospedalieri e/o sanitari, nell ultimo decennio sono stati isolati sempre più frequentemente, in numerosi altri ambienti e persino negli alimenti. La diffusione dell antibiotico resistenza tra gli enterococchi, noti come patogeni nosocomiali, ha contribuito a innalzarne il rischio di colonizzazione e infezione, incrementandone l importanza clinica. L'obiettivo principale della presente tesi di dottorato è stato quello di indagare la presenza e la struttura della popolazione enterococcica AR sia in matrici alimentari sia ambientali. In dettaglio, sono stati presi in considerazione due formaggi siciliani (Capitolo 1) e un impianto di depurazione urbano (Capitolo 2). Ad oggi, questa rappresenta la prima evidenza scientifica di AR in enterococchi isolati da formaggio Ragusano DOP e da un impianto di trattamento delle acque reflue urbane siciliano. I risultati ottenuti dal formaggio hanno evidenziato la presenza di una specifica popolazione, per lo più riferita alla specie E. durans, per la quale meno informazioni sull AR sono disponibili in letteratura. In particolare, è stata rilevata un elevata prevalenza di multi-resistenza e alcuni ceppi hanno evidenziato resistenza alla vancomicina. Inoltre, la distribuzione multimodale dei valori di MIC, all interno della specie, ha consentito la distinzione di due sottopopolazioni, suggerendo meccanismi di acquisizione genica di AR. Per quanto riguarda gli enterococchi isolati dal sistema di depurazione di acque urbane, i risultati hanno evidenziato la dominanza di E. faecium in tutti i siti analizzati e un'alta prevalenza di AR nei riguardi di numerosi antibiotici. Attraverso l analisi PFGE numerosi profili unici sono stati ottenuti e la presenza dello stesso clone con differenti profili di resistenza, in differenti siti, è stata osservata. In conclusione, i risultati ottenuti rafforzano la necessità di un monitoraggio costante e di una sorveglianza attiva sulla presenza di enterococchi AR sia nei formaggi che in ambienti acquatici, al fine di contenere il rischio per la salute umana

Comparative Bioavailability of Two Diosmin Formulations after Oral Administration to Healthy Volunteers

Diosmin is a flavonoid commonly found in citrus fruits, largely used as adjuvant treatment for circulatory disorders, including chronic venous insufficiency (CVI) and hemorrhoids. Following oral administration, diosmin is not directly absorbed but must first be hydrolyzed into its aglycone, diosmetin, which is then absorbed into the systemic circulation. The aim of the current cross-over clinical study was to assess the pharmacokinetic profile of µSmin® Plus, a micronized diosmin flavonoid complex standardized in diosmin and formulated with a buffering agent (tested formulation). The study compared this to unformulated micronized diosmin (reference), in 16 healthy volunteers. Plasma samples were analyzed by HPLC-MS and plasma diosmetin concentration was measured after deconjugation with β-glucuronidase. For the tested formulation area under the curve (AUC0-t), and maximum plasma and time concentration (Cmax; tmax) were found to be 298.4 ± 163.7, 50.3 ± 22.6 and 2.2 ± 2.9, respectively. AUC0-t and Cmax of the reference were 31.9 ± 100.4 and 2.4 ± 1.9, respectively. The tested formulation showed higher plasmatic concentrations of diosmetin in comparison to those obtained after the administration of unformulated micronized diosmin. The relative bioavailability was 9.4 greater for the tested formulation than in micronized diosmin. In conclusion, our data indicate that µSmin® Plus was rapidly and well absorbed into systemic circulation and may therefore be ideally suitable to deliver diosmin in human interventional trials