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Background: World Health Organization (WHO) has increasingly improved the guidelines to tackle the spread of Coronavirus Disease 2019 (COVID-19) among the worldwide population. In this context, each country has introduced specific social, healthcare, political and macroeconomic measures to face COVID pandemic locally. Objective: The general aim of this comparative overview is to highlight the most significant effects of COVID-19 pandemic on the main healthcare systems. Also, we critically analyzed the macroeconomic variables and the most promising solutions to improve both healthcare system and its related risk management, taking into specific consideration the most industrialized countries. Method: The main strategy has been built on a renewed concept of the hospital, rebuilding the old concepts of “triage” and “intensive care”. Recently, COVID-19 hospitals have allowed to cater the patients affected by COVID-19. Moreover, the reshaping of several healthcare policies and requirements has led to several positive effects, such as the recruitment of a huge number of human resources in the healthcare systems. Nevertheless, several negative effects have also impacted the communities mostly subjected to infections. Conclusion: Undoubtedly, the national healthcare systems have somehow addressed the people’s needs, trying not to neglect the social, healthcare, economic and political aspects. In our overview, we have reported how the different actions taken in the last months, have resulted in different outcomes.

It is generally held that the content of several free amino acids and dipeptides is closely related to the energy-supplying metabolism of skeletal muscles. Metabolic characteristics of muscles are involved in the variability of meat quality due to their ability to influence the patterns of energy metabolism not only in living animal but also during post-mortem time. Within this context, this study aimed at establishing whether the concentration of histidine dipeptides can affect muscle post-mortem metabolism, examining the glycolytic pathway of three chicken muscles (Pectoralis major, extensor iliotibialis lateralis and gastrocnemius internus as glycolytic, intermediate and oxidative-type, respectively) selected based on their histidine dipeptides content and ultimate pH. Thus, a total of 8 carcasses were obtained from the same flock of broiler chickens (Ross 308 strain, females, 49 days of age, 2.8 kg body weight at slaughter) and selected immediately after evisceration from the line of a commercial processing plant. Meat samples of about 1 cm3 were excised from bone-in muscles at 15, 60, 120 and 1,440 min post-mortem, instantly frozen in liquid nitrogen and used for the determination of pH, glycolytic metabolites, buffering capacity as well as histidine dipeptides content through 1H-NMR. Overall results suggest that glycolysis in leg muscles ceased already after 2 h post-mortem, while in breast muscle continued until 24 h, when it exhibited significantly lower pH values (P

Objectives: Vulvo-vaginal atrophy (VVA) is a highly prevalent chronic condition affecting the lives of postmenopausal women. Ospemifene and systemic hormone therapy (HT) improve vaginal health. This study aims to characterize the vaginal metabolic profile of women with VVA at baseline and after ospemifene and systemic HT. Study design: Sixty postmenopausal women, 32 of whom were affected by VVA, were consecutively enrolled. The vaginal metabolic profile of women with and without VVA at baseline and after three months of ospemifene or HT treatment was assessed. Main outcome measures: The following parameters were evaluated: (i) the Vaginal Health Index; (ii) the Vaginal Maturation Index; and (iii) vaginal metabolic profile, by means of 1H NMR spectroscopy. Results: The vaginal metabolome of postmenopausal women with VVA was different from that of postmenopausal women without VVA, including a more profound decrease in the levels of lactate and several amino acids, typically found in eubiosis, together with an enrichment of molecules derived from anaerobes and gut microbes. After 3 months, ospemifene and HT had modified the vaginal metabolome of the women with VVA, specifically by increasing the levels of beneficial molecules (e.g., lactate, leucine, glycine) and reducing those involved in dysbiosis (e.g., formate). HT improved the vaginal metabolome to a lesser extent. Conclusions: The vaginal metabolic profile of postmenopausal women with VVA differs from that of postmenopausal women without VVA. Our preliminary data show that both ospemifene and HT treatment increase the levels of molecules beneficial for vaginal health and reduce the levels of those involved in dysbiosis. HT improves the vaginal metabolic profile to a lower extent than ospemifene over the course of three months.

Mastitis is one of the diseases with the highest incidence in dairy cows, causing huge economic losses to the dairy industry all over the world. The aim of the study was to characterize mastitic milk metabolome through untargeted nuclear magnetic resonance spectroscopy (1H-NMR). Taking advantage of the high reproducibility of 1H-NMR, we had the opportunity to provide quantitative information for all the metabolites identified. Fifty-four molecules were characterized, sorted mainly into the chemical groups, namely amino acids, peptides and analogues, carbohydrates and derivates, organic acids and derivates, nucleosides, nucleotides and analogues. Combined with serum metabolomic investigations, several pathways were addressed to explain the mechanisms of milk metabolome variation affected by clinical mastitis, such as tricarboxylic acid cycle (TCA cycle) and phenylalanine, tyrosine and tryptophan biosynthesis. These results provide a further understanding of milk metabolome altered by clinical mastitis, which can be used as a reference for the further milk metabolome investigations.

During pregnancy, the vaginal ecosystem undergoes marked changes, including a significant enrichment with Lactobacillus spp. and profound alterations in metabolic profiles. A deep comprehension of the vaginal environment may shed light on the physiology of pregnancy and may provide novel biomarkers to identify subjects at risk of complications (e.g., miscarriage, preterm birth). In this study, we characterized the vaginal ecosystem in Caucasian women with a normal pregnancy (n = 64) at three different gestational ages (i.e., first, second and third trimester) and in subjects (n = 10) suffering a spontaneous first trimester miscarriage. We assessed the vaginal bacterial composition (Nugent score), the vaginal metabolic profiles (1H-NMR spectroscopy) and the vaginal levels of two cytokines (IL-6 and IL-8). Throughout pregnancy, the vaginal microbiota became less diverse, being mainly dominated by lactobacilli. This shift was clearly associated with marked changes in the vaginal metabolome: over the weeks, a progressive reduction in the levels of dysbiosis-associated metabolites (e.g., biogenic amines, alcohols, propionate, acetate) was observed. At the same time, several metabolites, typically found in healthy vaginal conditions, reached the highest concentrations at the end of pregnancy (e.g., lactate, glycine, phenylalanine, leucine, isoleucine). Lower levels of glucose were an additional fingerprint of a normal vaginal environment. The vaginal levels of IL-6 and IL-8 were significantly associated with the number of vaginal leukocytes, as well as with the presence of vaginal symptoms, but not with a condition of dysbiosis. Moreover, IL-8 concentration seemed to be a good predictor of the presence of vaginal Candida spp. Cytokine concentrations were negatively correlated to lactate, serine, and glycine concentrations, whereas the levels of 4-hydroxyphenyllactate, glucose, O-acetylcholine, and choline were positively correlated with Candida vaginal loads. Finally, we found that most cases of spontaneous abortion were associated with an abnormal vaginal microbiome, with higher levels of selected metabolites in the vaginal environment (e.g., inosine, fumarate, xanthine, benzoate, ascorbate). No association with higher pro-inflammatory cytokines was found. In conclusion, our analysis provides new insights into the pathophysiology of pregnancy and highlights potential biomarkers to enable the diagnosis of early pregnancy loss.

Vaginal microbes and their metabolic products have crucial functions, affecting local immunity development and maternal-fetal health. The composition of the vaginal microbiome can vary in response to various factors, including body mass index (BMI), and diet. In this study we get new insights into the vaginal ecosystem of Caucasian women (n = 24) at the first trimester of pregnancy, assessing whether pre-pregnancy diet can affect the structure of the vaginal environment in terms of bacterial composition and vaginal metabolite concentration. We characterized 1) the vaginal bacterial composition (Nugent score), 2) the vaginal metabolic profiles (1H-NMR spectroscopy), and 3) the dietary nutrient intake by means of a validated food frequency questionnaire. Pre-pregnancy BMI was negatively related to vaginal health status, indicating that women who begin pregnancy overweight/obese have a greater occurrence of vaginal dysbiosis during pregnancy. A lactobacilli-dominated vaginal microbiota was negatively associated with higher pre-pregnancy intake of animal-sourced protein. Conversely, a higher pre-pregnancy consumption of total carbohydrates and sugars seemed to be a protective factor for vaginal health. The vaginal environment of BV-women was characterized by higher levels of biogenic amines and organic acids, whereas higher levels of phenylpropionate and diverse amino acids were fingerprints of a healthy vaginal status. A significant association between a higher pre-pregnancy BMI and several dysbiosis-related vaginal metabolites was also found. Our study shed light on the role of pre-pregnancy BMI and diet on the vaginal environment during pregnancy, underlining the importance of limiting protein intake from animal foods to maintain a healthy lactobacilli-dominated microbiota.

Understanding the variations of muscle and plasma metabolites in response to high environmental temperature can provide important information on the molecular mechanisms related to body energy homeostasis in heat-stressed broiler chickens. In this study, we investigated the effect of chronic heat stress conditions on the breast muscle (Pectoralis major) and plasma metabolomics profile of broiler chickens by means of an innovative, high-throughput analytical approach such as the proton nuclear magnetic resonance (1H NMR) spectrometry. A total of 300 Ross 308 male chicks were split into two experimental groups and raised in either thermoneutral conditions for the entire rearing cycle (0–41 days) (TNT group; six replicates of 25 birds/each) or exposed to chronic heat stress conditions (30 °C for 24 h/day) from 35 to 41 days (CHS group; six replicates of 25 birds/each). At processing (41 days), plasma and breast muscle samples were obtained from 12 birds/experimental group and then subjected to 1H NMR analysis. The reduction of BW and feed intake as well as the increase in rectal temperature and heterophil: lymphocyte ratio confirmed that our experimental model was able to stimulate a thermal stress response without significantly affecting mortality. The 1H NMR analysis revealed that a total of 26 and 19 molecules, mostly related to energy and protein metabolism as well as antioxidant response, showed significantly different concentrations respectively in the breast muscle and plasma in response to the thermal challenge. In conclusion, the results obtained in this study indicated that chronic heat stress significantly modulates the breast muscle and plasma metabolome in fast-growing broiler chickens, allowing to delineate potential metabolic changes that can have important implications in terms of body energy homeostasis, growth performance and product quality.

Candida species are the most common cause of opportunistic fungal infections. Rapid identification and novel approaches for the characterization of these fungi are of great interest to improve the diagnosis and the knowledge about their pathogenic properties. This study aimed to characterize clinical isolates of Candida spp. by proteomics (MALDI-TOF MS) and metabolomics (1H-NMR), and to correlate their metabolic profiles with Candida species, source of infection and different virulence associated parameters. In particular, 49 Candida strains from different sources (blood, n = 15; vagina, n = 18; respiratory tract, n = 16), belonging mainly to C. albicans complex (61%), C. glabrata (20%) and C. parapsilosis (12%) species were used. Several extracellular and intracellular metabolites showed significantly different concentrations among isolates recovered from different sources of infection, as well as among different Candida species. These metabolites were mainly related to the glycolysis or gluconeogenesis, tricarboxylic acid cycle, nucleic acid synthesis and amino acid and lipid metabolism. Moreover, we found specific metabolic fingerprints associated with the ability to form biofilm, the antifungal resistance (i.e. caspofungin and fluconazole) and the production of secreted aspartyl proteinase. In conclusion, 1H-NMR-based metabolomics can be useful to deepen Candida spp. virulence and pathogenicity properties.

Background The use of an untargeted metabolomic approach to investigate biofluids of respiratory origin is of increasing interest in human and veterinary lung research. Considering the high incidence of equine asthma (> 14%) within horse population and the importance of this animal model for human disease, we aimed to investigate the metabolomic profile of bronchoalveolar lavage fluid (BALF) and exhaled breath condensate (EBC) in healthy and asthmatic horses. Results On the basis of clinical, endoscopic and BALF cytology findings, 6 horses with severe asthma (Group A) and 6 healthy horses (Group C) were included in the study. 1H-NMR analysis was used to identified metabolites in BALF and EBC samples. Metabolomic analysis allowed to identify and quantify 12 metabolites in BALF and seven metabolites in EBC. Among respiratory metabolites, myo-inositol, formate, glycerol and isopropanol in BALF, and methanol and ethanol in EBC, differed between groups (p Conclusions The application of metabolomic studies to investigate equine asthma using minimally invasive diagnostic methods, such as EBC metabolomics, provided promising results. According to our research, the study of selective profiles of BALF and EBC metabolites might be useful for identifying molecules like myo-inositol and methanol as possible biomarkers for airways diseases in horses.

Yak represents the main meat source for Tibetan people. This work aimed to investigate the metabolome of raw meat from Jiulong yaks, focusing on specimens farmed and harvested locally through traditional procedures. Untargeted nuclear magnetic resonance spectroscopy (1H-NMR) was selected as the analytical platform. Samples from longissimus thoracis, trapezius, triceps brachii and biceps femoris muscles, with different prevalences of red and white fibers, were selected. Among the fifty-three metabolites quantified in each of them, carnitine, carnosine, creatine and taurine are known for their bioactive properties. Twelve molecules were found to be differently concentrated in relation to muscle type. Longissimus thoracis, compared to biceps femoris, had higher concentrations of carnosine and formate and lower concentrations of mannose, inosine, threonine, IMP, alanine, valine, isoleucine, tyrosine, phenylalanine and leucine. A metabolic pathway analysis suggested that the main pathways differing among the muscles were connected to the turnover of amino acids. These results contribute to a deeper understanding of yak raw meat metabolism and muscle type differences, which can be used as an initial reference for the meat industry to set up muscle-specific investigations. The possibility of simultaneously quantifying several bioactive compounds suggests that these investigations could revolve around meat&rsquo, s nutritional value.

During the past few years, there has been an increasing prevalence of broiler breast muscle abnormalities, such as white striping (WS) and wooden breast conditions. More recently, a new muscular abnormality termed as spaghetti meat (SM) because of the altered structural integrity of the Pectoralis major muscle often associated with WS has emerged. Thus, this study aimed at evaluating the effects of WS and SM conditions, occurring alone or combined within the same P. major muscle, on meat quality traits and muscle histology. In two replications, 96 P. major muscles were classified into four classes: normal (N), WS, SM and WS/SM. The whole fillet was used for weight assessment and morphometric measurements, then each sample was cut in order to separate the superficial layer from the deep one and used to evaluate proximate composition, histological features, nuclear magnetic resonance relaxation times, functional properties and both myofibrillar and sarcoplasmic proteins profile. Fillets affected by WS and SM abnormalities exhibited higher weights and increased thickness and length. SM condition was associated with a relevant decrease in protein content coupled with a significant increase in moisture level, whereas fat content was affected only by the simultaneous presence of WS. Histological evaluations revealed that abnormal samples were characterized by several degenerative aspects that almost completely concerned the superficial layer of the fillets. White striped fillets exhibited necrosis and lysis of fibers, fibrosis, lipidosis, loss of cross striation and vacuolar degeneration. Moreover, SM samples were characterized by poor fiber uniformity and a progressive rarefaction of the endo- and peri-mysial connective tissue, whereas WS/SM fillets showed intermediate histological features. Nuclear magnetic resonance relaxation analysis revealed a higher proportion of extra-myofibrillar water in the superficial section of all the abnormal fillets, especially in SM samples, which consequently led to a reduction of the water holding capacity of meat. As for functional properties, abnormal fillets exhibited a lower protein solubility and higher ultimate pH values on both the superficial and deep sections. Although abnormal fillets exhibited higher yellowness values, no relevant effect on meat color was observed. The occurrence of WS and SM abnormalities led to increased carbonylation levels and more intense proteolytic processes. Overall, muscle abnormalities mainly affect the superficial layer of P. major muscle and particularly the occurrence of SM myopathy seems to implicate a more pronounced modification of meat quality traits than the mere presence of WS.

Autism spectrum disorders (ASD) make up a heterogeneous group of neurodevelopmental disorders characterized by social and communication difficulties associated with repetitive and restrictive behaviors. Besides core features, metabolic imbalances, inflammation, gastrointestinal (GI) symptoms, and altered gut microbiota composition were often described in association with ASD, but their connection with the severity of autism (SA) remains unexplored. In this study, fecal metabolome, microbiota, and calprotectin levels of 80 ASD preschoolers were quantified and correlated with SA. Twelve of the fifty-nine molecules that were quantified by fecal metabolome analysis were significantly associated with SA. No links between SA or GI symptoms and microorganisms’ relative abundance were highlighted. Significant correlations between bifidobacteria, Sutterella, lactobacilli relative abundance, and metabolomics profiles were found. These results suggest that fecal metabolome discriminates the SA and intestinal microorganisms mediate the link between metabolome and SA regardless of GI symptomatology. The study raises the possibility that grouping ASD populations through metabolomics and fecal microbiota could aid the identification of specific ASD endophenotypes, on the basis of the SA. Mechanistic studies focusing on detected biomarkers might be an option for future studies.

The aim of this work was to evaluate the effect of pulsed electric field (PEF) pre-treatment on mass transfer phenomena, water distribution and some physico-chemical parameters of osmo-dehydrated organic strawberries. For PEF treatments 100 near-rectangular shaped pulses, with fixed pulse width of 100 μs and repetition time of 10 ms were used. Electric fields strength applied were 100, 200 and 400 V cm−1. Afterwards, samples were subjected to OD treatments carried out in two different hypertonic solutions (40% w/w), one with sucrose and the other one with trehalose. The results shown that PEF treatment positively affected the mass transfer during OD even at the lowest electric field strength applied (100 V/cm), partially preserving the cell viability and maintaining at the same time the fresh-like characteristics of strawberries.

This work aimed to determine the effect of homogenization pressures (HPH) and addition of trehalose on the functional and technological properties of low pulp mandarin juice (LPJ). A set of experiments was designed, combining a non-targeted metabolomic 1 H NMR based approach together with suspended pulp and transmittance, hesperidin, vitamin C and antioxidant activity analysis. Suspended pulp increased with HPH and trehalose addition. Flavonoid hesperidin initially decreased with HPH but trehalose addition resulted in less flavonoid degradation during storage, increasing the effect with the HPH. Vitamin C was not affected by trehalose and pressure treatment but more Vitamin C degradation was observed in trehalose samples during storage. Antiradical activity improvement by trehalose was conditioned by homogenization pressures and specific bioactive compounds. 1 H NMR based approach highlighted the HPH effect on the microbiological aspects of low pulp mandarin juice by the identification of key molecules responsible of the microorganism profile evolution during storage.

To investigate the effects of oral bacteriotherapy on intestinal phenylalanine and tyrosine metabolism, in this longitudinal, double-arm trial, 15 virally suppressed HIV+ individuals underwent blood and fecal sample collection at baseline and after 6 months of oral bacteriotherapy. A baseline fecal sample was collected from 15 healthy individuals and served as control group for the baseline levels of fecal phenylalanine and tyrosine. CD4 and CD8 immune activation (CD38+) was evaluated by flow cytometry. Amino acid evaluation on fecal samples was conducted by Proton Nuclear Magnetic Resonance. Results showed that HIV+ participants displayed higher baseline phenylalanine/tyrosine ratio values than healthy volunteers. A significand reduction in phenylalanine/tyrosine ratio and peripheral CD4+ CD38+ activation was observed at the end of oral bacteriotherapy. In conclusion, probiotics beneficially affect the immune activation of HIV+ individuals. Therefore, the restoration of intestinal amino acid metabolism could represent the mechanisms through which probiotics exert these desirable effects.

Intense exercise causes to organisms to have oxidative stress and inflammation at the gastrointestinal (GI) level. The reduction in intestinal blood flow and the exercise-linked thermal damage to the intestinal mucosa can cause intestinal barrier disruption, followed by an inflammatory response. Furthermore, the adaptation to exercise may affect the gut microbiota and the metabolome of the biofluids. The aim of the present research was to evaluate the presence of a GI derangement in hunting dogs through a non-invasive sampling as a consequence of a period of intense exercise in comparison with samples collected at rest. The study included nine dogs that underwent the same training regime for hunting wild boar. In order to counterbalance physiological variations, multiple-day replicates were collected and pooled at each experimental point for each dog. The samples were collected immediately at rest before the training (T0), after 60 days of training (T1), after 60 days of hunting wild boar (T2), and finally, at 60 days of rest after hunting (T3). A number of potential stress markers were evaluated: fecal cortisol metabolites (FCMs) as a major indicator of altered physiological states, immunoglobulin A (IgA) as an indicator of intestinal immune protection, and total antioxidant activity [total antioxidant capacity (TAC)]. Since stool samples contain exfoliated cells, we investigated also the presence of some transcripts involved in GI permeability [occludin (OCLN), protease-activated receptor-2 (PAR-2)] and in the inflammatory mechanism [interleukin (IL)-8, IL-6, IL-1b, tumor necrosis factor alpha (TNFα), calprotectin (CALP), heme oxygenase-1 (HO-1)]. Finally, the metabolome and the microbiota profiles were analyzed. No variation in FCM and IgA content and no differences in OCLN and CALP gene expression between rest and training were observed. On the contrary, an increase in PAR-2 and HO-1 transcripts, a reduction in total antioxidant activity, and a different profile of microbiota and metabolomics data were observed. Collectively, the data in the present study indicated that physical exercise in our model could be considered a mild stressor stimulus.

Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) represent the most common agents of sexually transmitted rectal infections among men having sex with other men (MSM). In this study, we assessed the bacterial composition of the rectal microbiota associated with CT and/or NG infections in a cohort of men reporting unsafe rectal intercourse. A total of 125 rectal swabs were collected and four groups were compared: non-infected subjects (n = 53), patients with CT (n = 37), or NG rectal infection (n = 17) and patients with contemporary positivity for CT/NG (n = 18). CT and NG infections were detected by a real-time commercial test and the rectal microbiota composition was analyzed from rectal swabs through sequencing of the hypervariable V3-V4 regions of the 16S rRNA gene. The rectal microbiota of all subgroups was dominated by Prevotellaceae, Enterobacteriaceae, and Ruminococcaceae families. Irrespective of the analyzed subgroup, we found that the rectal environment of all the enrolled MSM was rich in Prevotella and Escherichia genera. Moreover, a shift in the bacterial composition between patients with sexually transmitted rectal infections and controls was noticed: infected patients were characterized by a depletion of Escherichia species, associated with an increase of anaerobic genera, including Peptoniphilus, Peptostreptococcus, and Parvimonas. Overall, the presence of rectal symptoms did not significantly modify the rectal microbiota profiles among the four groups of analyzed patients. We confirmed that HIV-positive patients are characterized by a lower bacterial richness than HIV-negative subjects. However, we found that the presence of HIV has a different impact on bacterial rectal communities compared to CT and NG infections, modifying the relative abundance of several genera, including Gardnerella, Lactobacillus, Corynebacterium, and Sutterella. Information about the rectal microbiota composition in CT and NG infections could shed light on the pathogenesis of these conditions and could contribute to the onset of new strategies for their control.

The vaginal microbiota plays a crucial role in maintaining the health and functioning of the female genital tract, preventing the colonization of urogenital pathogens and sexually transmitted infections. In this study, we characterized the vaginal bacterial communities and the metabolome associated to Chlamydia trachomatis infection (CT: 20 women), compared to healthy condition (H: 22 women) and bacterial vaginosis (BV: 19 women). A microarray-based tool (VaginArray), implemented with a real-time PCR for Gardnerella vaginalis, was used to determine the vaginal bacterial composition, whereas the metabolic profiles were assessed by a proton-based nuclear magnetic resonance (1H-NMR) spectroscopy. CT infection was characterized by bacterial and metabolic signatures similar to healthy condition, even though higher amounts of Lactobacillus iners, as well as depletion of some amino acids, biogenic amines, and succinate marked CT infection. Moreover, the frequency of Lactobacillus crispatus was higher in asymptomatic CT-positive patients than in women with CT-correlated symptoms. We also confirmed the marked differences in the microbiome and metabolome between healthy and BV-affected women. In conclusion, we highlighted microbial and metabolic peculiarities of the vaginal ecosystem in the case of CT infection, even though further studies are needed to understand if the observed alterations precede the infection onset or if the pathogen itself perturbs the vaginal environment.

none 13 no Published Online First: 12 September 2016 OBJECTIVE: The engagement of the gut microbiota in the development of symptoms and complications of diverticular disease has been frequently hypothesised. Our aim was to explore colonic immunocytes, gut microbiota and the metabolome in patients with diverticular disease in a descriptive, cross-sectional, pilot study. DESIGN: Following colonoscopy with biopsy and questionnaire phenotyping, patients were classified into diverticulosis or symptomatic uncomplicated diverticular disease; asymptomatic subjects served as controls. Mucosal immunocytes, in the diverticular region and in unaffected sites, were quantified with immunohistochemistry. Mucosa and faecal microbiota were analysed by the phylogenetic platform high taxonomic fingerprint (HTF)-Microbi.Array, while the metabolome was assessed by 1H nuclear magnetic resonance. RESULTS: Compared with controls, patients with diverticula, regardless of symptoms, had a >70% increase in colonic macrophages. Their faecal microbiota showed depletion of Clostridium cluster IV. Clostridium cluster IX, Fusobacterium and Lactobacillaceae were reduced in symptomatic versus asymptomatic patients. A negative correlation was found between macrophages and mucosal Clostridium cluster IV and Akkermansia. Urinary and faecal metabolome changes in diverticular disease involved the hippurate and kynurenine pathways. Six urinary molecules allowed to discriminate diverticular disease and control groups with >95% accuracy. CONCLUSIONS: Patients with colonic diverticular disease show depletion of microbiota members with anti-inflammatory activity associated with mucosal macrophage infiltration. Metabolome profiles were linked to inflammatory pathways and gut neuromotor dysfunction and showed the ability to discriminate diverticular subgroups and controls. These data pave the way for further large-scale studies specifically aimed at identifying microbiota signatures with a potential diagnostic value in patients with diverticular disease. Barbara, Giovanni; Scaioli, Eleonora; Barbaro, Maria Raffaella; Biagi, Elena; Laghi, Luca; Cremon, Cesare; Marasco, Giovanni; Colecchia, Antonio; Picone, Gianfranco; Salfi, Nunzio; Capozzi, Francesco; Brigidi, Patrizia; Festi, Davide Barbara, Giovanni; Scaioli, Eleonora; Barbaro, Maria Raffaella; Biagi, Elena; Laghi, Luca; Cremon, Cesare; Marasco, Giovanni; Colecchia, Antonio; Picone, Gianfranco; Salfi, Nunzio; Capozzi, Francesco; Brigidi, Patrizia; Festi, Davide

In the present study, data related to the metabolomics of saliva and serum in trained standardbred horses are provided for the first time. Metabolomic analysis allows to analyze all the metabolites within selected biofluids, providing a better understanding of biochemistry modifications related to exercise. On the basis of the current advances observed in metabolomic research on human athletes, we aimed to investigate the metabolites&rsquo, profile of serum and saliva samples collected from healthy standardbred horses and the relationship with physical exercise. Twelve trained standardbred horses were sampled for blood and saliva before (T0) and immediately after (T1) standardized exercise. Metabolomic analysis of both samples was performed by 1H-NMR spectroscopy. Forty-six metabolites in serum and 62 metabolites in saliva were detected, including alcohols, amino acids, organic acids, carbohydrates and purine derivatives. Twenty-six and 14 metabolites resulted to be significantly changed between T0 and T1 in serum and saliva, respectively. The findings of 2-hydroxyisobutyrate and 3-hydroxybutyrate in serum and GABA in equine saliva, as well as their modifications following exercise, provide new insights about the physiology of exercise in athletic horses. Glycerol might represent a novel biomarker for fitness evaluation in sport horses.

Recently, a certain number of broiler abattoirs located in different Countries around the World have signaled an emerging quality issue termed “gaping” because of the separation of the fiber bundles affecting the external portion of the bipinnate pectoralis minor muscle. Thus, after defining the criteria to classify the muscles as Normal (NORM), Moderate (MOD), or Severe (SEV) cases, the incidence of gaping under commercial conditions was assessed on a total of 8,600 P. minor obtained from broiler chickens belonging to 43 flocks during a 6-mo period. Then, a total of 180 P. minor were selected based on previously defined criteria to evaluate the main quality traits (pH, color, water-holding/-binding capacity and tenderness), proximate composition, water mobility, and thermal properties as well as metabolic profile through 1H-Nuclear Magnetic Resonance spectroscopy. The average incidence of gaping defect was found to be 16.8% (8.8 and 8.0% MOD and SEV cases, respectively). As for the main quality traits, a reduction in ultimate pH was observed as the severity of the gaping defect increased, with SEV muscles displaying significantly lower values in comparison with NORM (5.96 vs. 6.02; P < 0.01), while MOD showed intermediate values (5.99). Concurrently, if compared with their NORM counterpart, MOD and SEV exhibited higher lightness (53.6 and 54.2 vs. 51.8; P < 0.01) coupled with higher (P < 0.05) cooking losses and longer (P < 0.05) transversal relaxation time of extra-myofibrillar water fraction. Overall, no significant differences were found concerning proximate composition and thermal properties. With regard to the metabolic profile, a significantly lower (P < 0.001) glutamine concentration was found in MOD and SEV muscles that, concurrently, revealed significant (P < 0.05) variations in the metabolites involved in energy-generating pathways. Overall, these findings evidenced that the gaping defect affecting broilers’ P. minor muscles have strong similarities with the pale-soft-exudative condition previously described in poultry and likely results from the biochemical processes taking place during the post-mortem conversion of muscle to meat.

Background A wide debate is ongoing regarding the role of cutaneous dysbiosis in the pathogenesis and evolution of difficult-to-treat chronic wounds. Nowadays, probiotic treatment considered as an useful tool to counteract dysbiosis but the evidence in regard to their therapeutic use in the setting of difficult-to-treat cutaneous ulcers is still poor. Aim: clinical report An 83-year-old woman suffering a critical limb ischemia and an infected difficult-to-treat ulcerated cutaneous lesion of the right leg, was complementary treated with local application of a mixture of probiotic bacteria. Methods Microbiological and metabolomic analysis were conducted on wound swabs obtained before and after bacteriotherapy. Results During the treatment course, a progressive healing of the lesion was observed with microbiological resolution of the polymicrobial infection of the wound. Metabolomic analysis showed a significant difference in the local concentration of propionate, 2-hydroxyisovalerate, 2-oxoisocaproate, 2,3-butanediol, putrescine, thymine, and trimethylamine before and after bacteriotherapy. Conclusion The microbiological and metabolomic results seem to confirm the usefulness of complementary probiotic treatment in difficult-to-treat infected wounds. Further investigations are needed to confirm these preliminary findings.

Over the past several decades, as a result of the increasing growth rate and body size of modern hybrid birds, poultry industry has been facing up the occurrence of many breast meat abnormalities, such as White Striping (WS), Wooden Breast (WB) and Spaghetti Meat (SM), whose incidence has recently reached alarming levels. Thus, the present study aimed at simultaneously investigating the implications of WS, WB and SM abnormalities on meat colour, pH, proximate composition, fatty acid profile, collagen, thermal properties as well as texture and water mobility, assessed on both superficial and deep section of Pectoralis major muscle. Overall, the occurrence of breast abnormalities is associated with a higher ultimate pH and a significant increase in moisture and fat level, coupled with a decrease in protein and ash content with WB showing the most detrimental effect. However, fatty acid profile resulted to be mildly modified only by WS. Moreover, WB fillets showed significantly higher (P

In the quest for setting up rapid methods to evaluate water retention ability of meat microstructures, time domain nuclear magnetic resonance (TD-NMR) has gained a prominent role, due to the possibility to observe water located outside the myofibrils, easily lost upon storage or cooking. Diffusion weighted signals could be used to monitor the shape and dimension of the pores in which water is confined, thus boosting the information offered by TD-NMR. The work outlines a parsimonious model to describe relative abundance and diffusion coefficient of intra and extra myofibrillar water populations, exchange rate between them, diameter of the myofibrillar cells. To test our model, we registered diffusion and T2 weighted NMR signals at 20 MHz on fresh meat from pectoralis major muscle of 100 days old female turkey. We then purposely altered water distribution and myofibrils shape by means of freezing. The model predicted nicely the consequences of the imposed modifications.

In the past decades, the intense selection practices carried out in order to develop fast growing and high breast-yield turkey hybrids profoundly modified the muscle physiology leading to the development of growth-related alterations and muscular abnormalities. White striations of variable thickness have been particularly observed on the ventral surface of Pectoralis major muscle belonging from heavy male turkeys since several years. However, although the effects of white striping (WS) have been extensively studied on broilers, this condition was not considered as a main quality issue by both turkey producers and meat industry. Thus, this study aimed at evaluating whether the occurrence of WS in heavy male turkeys affects the quality traits and technological properties of meat to the same extent previously observed for broilers. In two replications, 72 Pectoralis major muscles were classified as: normal (NORM), moderate WS (MOD) and severe WS (SEV) cases. The whole muscle was weighed and cut in order to assess colour, ultimate pH, water holding (drip and cooking losses) and binding (marinade uptake) capacities, NMR relaxation properties, shear force as well as proximate composition of meat. The Pectoralis major muscles affected by WS (both moderate and severe cases) exhibited a one-fifth increased weight in comparison with their NORM counterpart. However, the occurrence of WS only partially affected the proximate composition of the meat. In detail, although moisture, collagen and protein contents did not differ among the groups, if compared with NORM, higher lipid levels were found in SEV muscles, whereas MOD had intermediate values. On the other hand, both MOD and SEV exhibited lower ash content. Despite these variations in proximate composition, both water holding and binding capacities of turkey breast meat were not affected by WS. Indeed, quality traits of raw (pH, colour, cooking losses and shear force) and marinated (uptake, cooking losses and shear force) meat as well as water distribution within the muscle tissue did not differ between NORM and WS cases. Overall, if compared with broilers, WS only marginally affected quality traits of turkey breast meat. It might thus be hypothesised a diverse specie-specific physiological response to the pressure in muscle tissue induced by the selection in turkeys that, although analogously led to the occurrence of WS, results in limited effects on meat quality.

The attention of sports community toward probiotic supplementation as a way to promote exercise and training performance, together with good health, has increased in recent years. This has applied also to horses, with promising results. Here, for the first time, we tested a probiotic mix of several strains of live bacteria typically employed for humans to improve the training performance of Standardbred horses in athletic activity. To evaluate its effects on the horse performance, we measured lactate concentration in blood, a translational outcome largely employed for the purpose, combined with the study of hematological and biochemical parameters, together with urine from a metabolomics perspective. The results showed that the probiotic supplementation significantly reduced postexercise blood lactate concentration. The hematological and biochemical parameters, together with urine molecular profile, suggested that a likely mechanism underlying this positive effect was connected to a switch of energy source in muscle from carbohydrates to short-chain fatty acids. Three sulfur-containing molecules differently concentrated in urines in connection to probiotics administration suggested that such switch was linked to sulfur metabolism. NEW & NOTEWORTHY Probiotic supplementation could reduce postexercise blood lactate concentration in Standardbred horses in athletic activity. Blood parameters, together with urine molecular profile, suggest the mechanism underlying this positive effect is connected to a switch of energy source in muscle from carbohydrates to short-chain fatty acids. Sulfur-containing molecules found in urines in connection to probiotics administration suggested that such switch was linked to sulfur metabolism.

International audience; In vitro alternatives to clinical trials are used for studying human food digestion. For simulating infant digestion, only few models, lacking of physiological relevance, are available. Thanks to an extensive literature review of the in vivo infant digestive conditions, a gastrointestinal static in vitro model was developed for infants born at term and aged of 28 days. The model was applied to the digestion of a commercial infant formula. Kinetics of digestion, as well as the structural evolution, were compared with those obtained while submitting the same formula to the adult international consensus protocol of in vitro static digestion. The kinetics of proteolysis and lipolysis differed according to the physiological stage resulting mainly from the reduced level of enzymes and bile salts as well as the higher gastric pH in the infant model. This in vitro static model of infant digestion is of interest for scientists, food or pharmaceutical manufacturers.

The present work characterized the metabolomic profile of tracheal wash (TW) and exhaled breath condensate (EBC) in healthy horses and horses with respiratory disease. Six asthma-affected horses (group A) and six healthy controls (group H) underwent clinical, endoscopic and cytologic examinations of upper airways to confirm the active phase of asthma. TW and EBC samples were collected from each animal and investigated by proton nuclear magnetic resonance (H-1-NMR) metabolomic analysis. A total of ten out of 38 metabolites found in the TW were significantly different between the groups (p < 0.05). Higher concentrations of histamine and oxidant agents, such as glutamate, valine, leucine and isoleucine, as well as lower levels of ascorbate, methylamine, dimethylamine and O-phosphocholine, were found in group A compared to group H. Eight metabolites were found in equine EBC, namely methanol, ethanol, formate, trimethylamine, acetone, acetate, lactate and butanone, previously observed also in human EBC. Despite the fact that this was a pilot study, the results showed that the metabolomic analysis of TW and EBC has the potentiality to serve as a basis for diagnostic tools in horses with asthma.

The optimal application of Pulsed Electric Fields (PEF) technology, used by food industry to assist mass transfer processes, depends on the effectiveness of the induced electroporation. The present work aimed at exploring the application of Magnetic Resonance Imaging (MRI) combined with Computer Vision System (CVS) analysis to assess the spatial distribution of electroporation in apple tissue. PEF-treated apple samples were compared with Dipping (Dip) and Vacuum Impregnation (VI) to gain insight into the spatial distribution of mechanisms that lead to microstructural modifications over time. CVS showed that electroporation modified heterogeneously apple microstructure, causing enzymatic browning unevenly across the samples. MRI transverse relaxation times (T2) maps and longitudinal relaxation times (T1)-weighted images throughout apple tissue confirmed the inhomogeneous distribution and extent of the cell disruption, along with the release of intracellular content toward the external solution. Industrial relevance The novel applications of pulsed electric fields require fast and reliable methods to detect and estimate the breakage of the membranes integrity in order to boost their industrial adoption and optimization. The present study provided analytical tools able to monitor the spatial distribution of electroporation in plant tissue samples within minutes and consequently to speed up and improve the assessment of different PEF treatments.

Objective Effect of a probiotic on the gut microbiome and peripheral immune function in healthy controls and relapsing-remitting multiple sclerosis (MS) patients. Methods MS patients (N = 9) and controls (N = 13) were orally administered a probiotic containing Lactobacillus, Bifidobacterium, and Streptococcus twice-daily for two months. Blood and stool specimens were collected at baseline, after completion of the 2-month treatment, and 3 months after discontinuation of therapy. Frozen peripheral blood mononuclear cells (PBMCs) were used for immune cell profiling. Stool samples were used for 16S rRNA profiling and metabolomics. Results Probiotic administration increased the abundance of several taxa known to be depleted in MS such as Lactobacillus. We found that probiotic use decreased the abundance of taxa previously associated with dysbiosis in MS, including Akkermansia and Blautia. Predictive metagenomic analysis revealed a decrease in the abundance of several KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways associated with altered gut microbiota function in MS patients, such as methane metabolism, following probiotic supplementation. At the immune level, probiotic administration induced an anti-inflammatory peripheral immune response characterized by decreased frequency of inflammatory monocytes, decreased mean fluorescence intensity (MFI) of CD80 on classical monocytes, as well as decreased human leukocyte antigen (HLA) D related MFI on dendritic cells. Probiotic administration was also associated with decreased expression of MS risk allele HLA-DQA1 in controls. Probiotic-induced increase in abundance of Lactobacillus and Bifidobacterium was associated with decreased expression of MS risk allele HLA.DPB1 in controls. Interpretation Our results suggest that probiotics could have a synergistic effect with current MS therapies. Ann Neurol 2018.

Introduction: To investigate the effectiveness and the safety of a probiotic-mixture (Vivomixx®, Visbiome®, DeSimone Formulation®; Danisco-DuPont, Madison, WI, USA) for the treatment of infantile colic in breastfed infants, compared with a placebo. Methods: A randomized, double-blind, placebo-controlled trial was conducted in exclusively breastfed infants with colic, randomly assigned to receive a probiotic-mixture or a placebo for 21 days. A structured diary of gastrointestinal events of the infants was given to the parents to complete. Samples of feces were also collected to evaluate microbial content and metabolome using fecal real-time polymerase chain reaction (qPCR) and Nuclear magnetic resonance (NMR)-based analysis. Study registered at ClinicalTrials.gov (NCT01869426). Results: Fifty-three exclusively-breastfed infants completed three weeks of treatment with a probiotic-mixture (n = 27) or a placebo (n = 26). Infants receiving the probiotic-mixture had less minutes of crying per day throughout the study by the end of treatment period (68.4 min/day vs. 98.7 min/day; p = 0.001). A higher rate of infants from the probiotic-mixture group responded to treatment (defined by reduction of crying times of ≥50% from baseline), on day 14, 12 vs. 5 (p = 0.04) and on day 21, 26 vs. 17 (p = 0.001). A higher quality of life, assessed by a 10-cm visual analogue scale, was reported by parents of the probiotic-mixture group on day 14, 7.1 ± 1.2 vs. 7.7 ± 0.9 (p = 0.02); and on day 21, 6.7 ± 1.6 vs. 5.9 ± 1.0 (p = 0.001). No differences between groups were found regarding anthropometric data, bowel movements, stool consistency or microbiota composition. Probiotics were found to affect the fecal molecular profile. No adverse events were reported. Conclusions: Administration of a probiotic-mixture appears safe and reduces inconsolable crying in exclusively breastfed infants.

Item does not contain fulltext Diminished colonic health is associated with various age-related pathologies. Calorie restriction (CR) is an effective strategy to increase healthy lifespan, although underlying mechanisms are not fully elucidated. Here, we report the effects of lifelong CR on indicators of colonic health in aging C57Bl/6J mice. Compared to an ad libitum control and moderate-fat diet, 30% energy reduction was associated with attenuated immune- and inflammation-related gene expression in the colon. Furthermore, expression of genes involved in lipid metabolism was higher upon CR, which may point towards efficient regulation of energy metabolism. The relative abundance of bacteria considered beneficial to colonic health, such as Bifidobacterium and Lactobacillus, increased in the mice exposed to CR for 28 months as compared to the other diet groups. We found lower plasma levels of interleukin-6 and lower levels of various metabolites, among which are bile acids, in the colonic luminal content of CR-exposed mice as compared to the other diet groups. Switching from CR to an ad libitum moderate-fat diet at old age (24 months) revealed remarkable phenotypic plasticity in terms of gene expression, microbiota composition and metabolite levels, although expression of a subset of genes remained CR-associated. This study demonstrated in a comprehensive way that CR affects indicators of colonic health in aging mice. Our findings provide unique leads for further studies that need to address optimal and feasible strategies for prolonged energy deprivation, which may contribute to healthy aging.

Background: In addition to strain taxonomy, the ability of probiotics to confer beneficial effects on the host rely on a number of additional factors including epigenetic modulation of bacterial genes leading to metabolic variability and might impact on probiotic functionality.Aims: To investigate metabolism and functionality of two different batches of a probiotic blend commercialized under the same name in Europe in models of intestinal inflammation.Methods: Boxes of VSL#3, a probiotic mixture used in the treatment of pouchitis, were obtained from pharmacies in UK subjected to metabolomic analysis and their functionality tested in mice rendered colitis by treatment with DSS or TNBS.Results: VSL#3-A (lot DM538), but not VSL#3-B (lot 507132), attenuated “clinical” signs of colitis in the DSS and TNBS models. In both models, VSL#3-A, but not VSL#3-B, reduced macroscopic scores, intestinal permeability, and expression of TNFα, IL-1β, and IL-6 mRNAs, while increased the expression of TGFβ and IL-10, occludin, and zonula occludens-1 (ZO-1) mRNAs and shifted colonic macrophages from a M1 to M2 phenotype (P < 0.05 vs. TNBS). In contrast, VSL#3-B failed to reduce inflammation, and worsened intestinal permeability in the DSS model (P < 0.001 vs. VSL#3-A). A metabolomic analysis of the two formulations allowed the identification of two specific patterns, with at least three-folds enrichment in the concentrations of four metabolites, including 1–3 dihydroxyacetone (DHA), an intermediate in the fructose metabolism, in VSL#3-B supernatants. Feeding mice with DHA, increased intestinal permeability.Conclusions: Two batches of a commercially available probiotic show divergent metabolic activities. DHA, a product of probiotic metabolism, increases intestinal permeability, highlighting the complex interactions between food, microbiota, probiotics, and intestinal inflammation.

Introduction In healthy women, lactobacilli play a crucial role in maintaining the microbial homeostasis of the vaginal niche. In case of bacterial vaginosis (BV), a condition characterised by a depletion of lactobacilli and an increasing number of anaerobes, a higher risk of urogenital and sexually transmitted infections (STIs) is reported. The vaginal environment of healthy and BV-positive women have been extensively studied, leading to the identification of the microbial species dominating these opposite conditions and to the description of specific metabolic profiles. Besides that, less is known about the vaginal microbiome in case of STIs, as Chlamydia trachomatis (CT) infections. The aim of this study was to analyse the composition of the endogenous microbiota and the metabolic signatures of the vaginal niche in 3 different conditions: healthy, BV and CT infections. Methods From July 2016, all the pre-menopausal women attending the STI Outpatients Clinic of Sant’Orsola-Malpighi Hospital in Bologna (Italy) and meeting one of the following criteria were enrolled: presence of vaginal symptoms or presence of risk factors for CT infection. Patients with vaginal candidiasis were excluded. For all the patients, a vaginal swab was collected for molecular CT detection (Versant CT/GC DNA 1.0 Assay; Siemens), whereas Amsel criteria were used for BV assessment. Moreover, for each woman, an additional vaginal swab stored in saline was collected and centrifuged. Cell pellets were examined with a DNA-microarray platform including 17 probe sets specific for the most representative vaginal bacterial groups and with a quantitative real-time PCR targeting 16s rRNA gene of Gardnerella vaginalis (GV). Cell-free supernatants were used for metabolomic analysis by means of 1 H-NMR spectroscopy. NMR spectra were recorded with an AVANCE spectrometer (Bruker). Similarities among microbial and metabolic profiles of samples were investigated by means of a principal component analysis (PCA). Differences in GV DNA loads and metabolites concentrations were analysed by ANOVA test. The study was approved by the Hospital Ethical Committee. Results Among all the women enrolled, 25 were considered healthy, 18 received a diagnosis of BV and 22 were positive for CT. PCA revealed that the vaginal microbiome of healthy and BV-subjects were clearly distinct and that CT-positive women were more similar to healthy women rather than to BV-positives, both for microbial composition and for metabolic profile. The mean GV DNA load was significantly different between the groups (p=0.03): healthy and CT positive women showed similar and lower mean loads compared to BV group. At a metabolic level, significantly higher concentrations of formate, ethanolamine and methylamine were found in BV-patients, while tryptophan and lactate were more present in healthy and CT-positive women. Conclusion Specific microbial and metabolic signatures characterise different clinical conditions of the vaginal tract. In this context, CT-positive women are definitely more similar to healthy than BV-subjects.

The aim of this study was to investigate the influence of oral iron supplementation, in the form of fortified breads, on the growth performance, health, iron status parameters, and fecal metabolome of anemic piglets. A study was conducted on 24 hybrid (Large White × Landrace × Duroc) piglets. From day 44, the post-natal 12 piglets were supplemented with 100 g of one of two experimental breads, each fortified with 21 mg of ferrous sulphate, either encapsulated or not. After one week of oral supplementation, hematological parameters (hematocrit value, hemoglobin, and red blood cells) showed statistically significant differences (p ≤ 0.05). Piglets fed with the fortified breads had higher iron concentrations in the heart, liver, and intestinal mucosa compared to anemic piglets fed with control bread. Gene expression of hepcidin, iron exporter ferroportin (IREG1), and divalent metal transporter 1 (DMT1), together with concentrations of plasma ferritin, showed no significant statistical differences between groups. Both fortified breads could be used as sources of bioavailable iron. The seven-day intervention trial showed microencapsulation to have only a mild effect on the effectiveness of iron supplementation in the form of fortified bread.

The feasibility of enriching fish cakes with long chain omega-3 polyunsaturated fatty acids (n-3 LC-PUFAs) as a source of high quality lipids and seaweed extracts as a natural antioxidant was investigated. The LC-PUFAs were in the form of bulk cod liver oil and microencapsulated fish oil. The sensory quality and lipid oxidation were studied by means of chemical analysis, namely peroxide value (PV) and thiobarbituric acid reactive substances (TBARS), and sensory evaluation (descriptive analysis), throughout 28-days of refrigerate storage. In addition near-infrared (NIR) spectroscopy was assessed as a rapid tool to evaluate TBARS content. Omega-3 enriched samples showed slightly higher levels of lipid oxidation than the control samples but microencapsulation of fish oil seemed to protect PUFAs, leading to a lower content of peroxides and a lower rate of formation of secondary compounds during storage. Addition of seaweed extracts did not affect lipid oxidation nor the quality of the products as evaluated with descriptive analysis. No off-flavor was detected in any of the samples and low scores in rancid odor and flavor were registered.