Metagenetic and Volatilomic Approaches to Elucidate the Effect of Lactiplantibacillus plantarum Starter Cultures on Sicilian Table Olives

The present study aimed to evaluate the effect of selected Lactiplantibacillus plantarum strains on both microbiota composition and volatile organic compound profile of Sicilian table olives. Two mixed cultures, named O1 and O2, were set up for pilot-plan scale fermentations at 5% of NaCl. Uninoculated table olives at 5 and 8% (C5 and C8) of salt were used as control. The fermentation process was monitored until 80 days through a dual approach, which included both classical microbiological and 16S amplicon-based sequencing and volatilomics analyses. Compared with control samples (C5 and C8), experimental samples, inoculated with starter cultures (O1 and O2), exhibited a faster acidification with a more pronounced drop in pH. Metagenetics data revealed significant differences of microbiota composition among samples, highlighting the dominance of lactobacilli in both experimental samples; a high occurrence of Enterobacter genus only in control samples with 5% of NaCl; and the presence of Bacteroides, Faecalibacterium, Klebsiella, and Raoultella genera only in control samples with 8% of NaCl. Furthermore, microbiota composition dynamics, through the fermentation process, significantly affected the volatile organic compounds of the final products, whereas no compounds involved in off-odors metabolites were detected in all samples investigated. In conclusion, the addition of the proposed starter cultures and the use of low concentrations of sodium chloride positively affected the microbiota and volatile organic compounds, ensuring the microbiological safety and the pleasant flavors of the final product

FoodOmics as a new frontier to reveal microbial community and metabolic processes occurring on table olives fermentation

Table olives are considered the most widespread fermented food in the Mediterranean area and their consumption is expanding all over the world. This fermented vegetable can be considered as a natural functional food thanks to their high nutritional value and high content of bioactive compounds that contribute to the health and well-being of consumers. The presence of bioactive compounds is strongly influenced by a complex microbial consortium, traditionally exploited through culture-dependent approaches. Recently, the rapid spread of omics technologies has represented an important challenge to better understand the function, the adaptation and the exploitation of microbial diversity in different complex ecosystems, such as table olives. This review provides an overview of the potentiality of omics technologies to in depth investigate the microbial composition and the metabolic processes that drive the table olives fermentation, affecting both sensorial profile and safety properties of the final product. Finally, the review points out the role of omics approaches to raise at higher sophisticated level the investigations on microbial, gene, protein, and metabolite, with huge potential for the integration of table olives composition with functional assessments

Effects of different stress parameters on growth and on oleuropein-degrading abilities of lactiplantibacillus plantarum strains selected as tailored starter cultures for naturally table olives

The use of β-glucosidase positive strains, as tailored-starter cultures for table olives fermentation, is a useful biotechnological tool applied to accelerate the debittering process. Nowadays, strains belonging to Lactiplantibacillus plantarum species are selected for their high versatility and tolerance to stress conditions. The present study investigated the effect of different stress factors (pH, temperature and NaCl) on growth and on oleuropein-degrading abilities of selected L. plantarum strains. In addition, the presence of the beta-glucosidase gene was investigated by applying a PCR based approach. Results revealed that, overall, the performances of the tested strains appeared to be robust toward the different stressors. However, the temperature of 16 °C significantly affected the growth performance of the strains both singularly and in combination with other stressing factors since it prolongs the latency phase and reduces the maximum growth rate of strains. Similarly, the oleuropein degradation was mainly affected by the low temperature, especially in presence of low salt content. Despite all strains displayed the ability to reduce the oleuropein content, the beta-glucosidase gene was detected in five out of the nine selected strains, demonstrating that the ability to hydrolyze the oleuropein is not closely related to the presence of beta-glucosidase. Data of the present study suggest that is extremely important to test the technological performances of strains at process conditions in order to achieve a good selection of tailored starter cultures for table olives

Levels of l-arginine and l-citrulline in patients with erectile dysfunction of different etiology

Nitric oxide is a physiologic signal essential to penile erection. l-citrulline (l-Cit) is converted into l-arginine (l-Arg), the precursor from which nitric oxide is generated. The level of l-Arg and l-Cit in the field of male sexual function remains relatively underexplored. The aim of the study was to evaluate the level of serum l-Arg and of l-Cit in a group of patients with erectile dysfunction. Diagnosis and severity of erectile dysfunction was based on the IIEF-5 and its etiology was classified as arteriogenic (A-ED), borderline (BL-ED), and non-arteriogenic (NA-ED) with penile echo-color-Doppler in basal condition and after intracaversous injection of prostaglandin E1. Serum l-Arg and l-Cit concentrations were measured by a cation-exchange chromatography system. l-Arg and l-Cit levels of men with A-ED were compared with those of male with BL-ED and NA-ED. Median level of l-Arg and l-Cit in 122 erectile dysfunction patients (41 A-ED, 23 ED-BL, 58 NA-ED) was 82.7 and 35.4\ua0\u3bcmol/L, respectively. l-Arg and l-Cit levels in control patients were not significantly different (p\ua0=\ua00.233 and p\ua0=\ua00.561, respectively) than in total erectile dysfunction patients. l-Arg and l-Cit levels in control patients were significantly higher (p\ua0\ua00.50) was observed in controls and in both BL-ED and NA-ED patients. Patients with severe/complete-erectile dysfunction (IIEF-5\ua0\ua00.3) but without reaching the statistical significance. Our study shows that a significant proportion of erectile dysfunction patients have low l-Arg or l-Cit level and that this condition is more frequent in patients with arteriogenic etiology. Low levels of these nitric oxide synthase substrates might increase the erectile dysfunction risk by reducing the concentration of nitric oxide

ESX1 gene expression as a robust marker of residual spermatogenesis in azoospermic men

Background It would be of value to identify ongoing spermatogenesis molecular markers which can predict successful sperm recovery in patients with non-obstructive azoospermia undergoing conventional or microsurgical testicular sperm extraction (TESE/microTESE). ESX1 is an X-linked homeobox gene expressed in testis, placenta, brain and lung in humans and specifically in pre-and post-meiotic germ cells of the testis in mice. Methods We investigated the sequence, expression (by RT-PCR) and epigenetic status (by promoter pyrosequencing) of ESX1 in testicular tissue samples, obtained from 81 azoospermic subjects in the context of surgical sperm extraction, to check a possible association between ESX1 alterations and impaired spermatogenesis, as determined by histological analysis. Result SThe ESX1 transcript was detected in 100 of cases diagnosed as obstructive azoospermia (33), hypospermatogenesis (18) and incomplete maturation arrest (MA) (2), and sperm recovery was also successful in 100 of these cases. ESX1 mRNA was also detected in 5 of 6 patients with incomplete Sertoli cell-only syndrome, in 4 of 6 subjects with complete MA but in only 3 of 16 cases of complete Sertoli cell-only syndrome (cSCOS), whereas sperm recovery was successful in 4 of 6, 2 of 6 and 5 of 16 of these patients, respectively. In cases of focal spermatogenesis, ESX1 expression and sperm retrieval were concordant in 14 of 19 (74) cases subjected to TESE, but in only 3 of 11 (27) men who underwent microTESE. With TESE, but not with microTESE, both samples originated from adjacent testicular areas. The pyrosequencing of the ESX1 CpG island revealed methylation levels that were significantly lower in ESX1 expressors when compared with non-expressors. Conclusions ESX1 emerges as a potentially reliable spermatogenesis molecular marker, whose clinical value as a predictor of successful sperm retrieval warrants further studies