Score plot and loading plot of the first and secondary principal components based on area and number peaks obtained from reverse phase high pressure liquid chromatography (RP-HPLC).

<p>Score plot and loading plot (panels 1, 3, 5) of the first and secondary principal components (PC) (panels 2, 4, 6) after PC analysis based on area and number peaks obtained from reversed-phase high-protein liquid chromatograms of the pH 4.6-soluble fractions of Fiore Sardo (A), Pecorino Siciliano (B) and Pecorino Toscano (C). Nine sub-blocks are identified by the letters A—I. Sub-blocks A, D, and G, and sub-blocks C, F and I were collected from top and bottom surface region, respectively, whereas sub-blocks B and H from inner side region, and sub-block E from the core. The whole slice was the control Control. Further details were reported in the Material and Methods and in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0153213#pone.0153213.g001" target="_blank">Fig 1</a>.</p

Concentrations of total free amino acids (FAA) (mg/kg) identified in Fiore Sardo, Pecorino Siciliano and Pecorino Toscano cheeses.

<p>Nine sub-blocks are identified by the letters A—I. Sub-blocks A, D, and G, and sub-blocks C, F and I were collected from top and bottom surface region, respectively, whereas sub-blocks B and H from inner side region, and sub-block E from the core. The whole slice was the control Control. Further details were reported in the Material and Methods and in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0153213#pone.0153213.g001" target="_blank">Fig 1</a>.</p

Operational taxonomic unit (OTUs) occurring at 0.1% abundance in at least one sample and related number, assigned to the species level when such assignment was possible of Fiore Sardo, Pecorino Siciliano and Pecorino Toscano cheeses.

<p>Operational taxonomic unit (OTUs) occurring at 0.1% abundance in at least one sample and related number, assigned to the species level when such assignment was possible of Fiore Sardo, Pecorino Siciliano and Pecorino Toscano cheeses.</p

Correlations between the abundance of operational taxonomic units (OTUs) and proteolysis and synthesis of volatile components.

<p>Correlations between the abundance of OTUs and total free amino acid (FFA), concentration of Ser, Glu, Asp, and Arg, number of hydrophilic peaks of the pH 4.6-soluble nitrogen fractions, and volatile components (arbitrary units of area) identified from Fiore Sardo, Pecorino Siciliano, and Pecorino Toscano cheeses. Euclidean distance and McQuitty’s criterion (weighted pair group method with averages) were used for clustering. The colors correspond to normalized mean data levels from low (grey) to high (yellow). The color scale, in terms of units of standard deviation, is shown at the top. Only the positive correlations with a P<0.05, FDR<0.05 and r>0.7 are reported. <i>Lactococcus lactis</i>, Lc. lactis; <i>Streptococcus thermophilus</i>, Sc. thermophilus; <i>Staphylococcus equorum</i>, St. equorum; <i>Lactobacillus plantarum</i>, Lb. plantarum; <i>Lactobacillus brevis</i>, Lb. brevis; <i>Lactobacillus coryniformis</i>, Lb. coryniformis; <i>Lactobacillus helveticus</i>, Lb. helveticus; <i>Lactobacillus delbrueckii</i>, Lb. delbruekii; <i>Lactobacillus</i> sp., Lb. sp; <i>Lactobacillus crustorum</i>, Lb. crustorum; <i>Lactobacillus buchneri</i>, Lb. buchneri; <i>Lactobacillus parabuchneri</i>, Lb. parabuchneri; <i>Brevibacterium</i> sp., B; <i>Brachybacterium</i> sp., Br; <i>Halomonas variabilis</i>, H. variabilis; Art, <i>Arthrobacter</i>.</p

Cell numbers (log CFU/g)^* of various microbial groups in Fiore Sardo, Pecorino Siciliano and Pecorino Toscano cheeses.

<p>Cell numbers (log CFU/g)<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0153213#t001fn002" target="_blank">^*</a> of various microbial groups in Fiore Sardo, Pecorino Siciliano and Pecorino Toscano cheeses.</p

Abundance and number of operational taxonomic units (OTUs) assigned at genus level occurring in Fiore Sardo, Pecorino Siciliano, and Pecorino Toscano cheeses.

<p>Distribution of the OTUs assigned at genus level occurring in Fiore Sardo (A), Pecorino Siciliano (B), and Pecorino Toscano (C) cheese. Nine sub-blocks are identified by the letters A—I. Sub-blocks A, D, and G, and sub-blocks C, F and I were collected from top and bottom surface region, respectively, whereas sub-blocks B and H from inner side region, and sub-block E from the core. The whole slice was the control. Further details were reported in the Material and Methods and in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0153213#pone.0153213.g001" target="_blank">Fig 1</a>.</p

Image_1_Fermentation of Nocellara Etnea Table Olives by Functional Starter Cultures at Different Low Salt Concentrations.TIF

<p>Nocellara Etnea is one of the main Sicilian cultivars traditionally used to produce both olive oil and naturally fermented table olives. In the present study, the effect of different salt concentrations on physico-chemical, microbiological, sensorial, and volatile organic compounds (VOCs) formation was evaluated in order to obtain functional Nocellara Etnea table olives. The experimental design consisted of 8 treatments as follow: fermentations at 4, 5, 6, and 8% of salt with (E1-E4 samples) and without (C1-C4 samples) the addition of starters. All the trials were carried out at room temperature (18 ± 2°C) and monitored for an overall period of 120 d. In addition, the persistence of the potential probiotic Lactobacillus paracasei N24 at the end of the process was investigated. Microbiological data revealed the dominance of lactic acid bacteria (LAB), starting from the 7th d of fermentation, and the reduction of yeasts and enterobacteria in the final product inoculated with starters. VOCs profile highlighted a high amount of aldehydes at the beginning of fermentation, which significantly decreased through the process and a concomitant increase of alcohols, acids, esters, and phenols. In particular, esters showed an occurrence percentage higher in experimental samples rather than in control ones, contributing to more pleasant flavors. Moreover, acetic acid, ethanol, and phenols, which often generate off-flavors, were negatively correlated with mesophilic bacteria and LAB. It is interesting to note that salt content did not affect the performances of starter cultures and slightly influenced the metabolome of table olives. Sensory data demonstrated significant differences among samples registering the highest overall acceptability in the experimental sample at 5% of NaCl. The persistence of the L. paracasei N24 strain in experimental samples, at the end of the process, revealed its promising perspectives as starter culture for the production of functional table olives with reduced salt content.</p

Fecal Microbiota and Metabolome of Children with Autism and Pervasive Developmental Disorder Not Otherwise Specified

<div><p>This study aimed at investigating the fecal microbiota and metabolome of children with Pervasive Developmental Disorder Not Otherwise Specified (PDD-NOS) and autism (AD) in comparison to healthy children (HC). Bacterial tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) of the 16S rDNA and 16S rRNA analyses were carried out to determine total bacteria (16S rDNA) and metabolically active bacteria (16S rRNA), respectively. The main bacterial phyla (Firmicutes, Bacteroidetes, Fusobacteria and Verrucomicrobia) significantly (<i>P</i><0.05) changed among the three groups of children. As estimated by rarefaction, Chao and Shannon diversity index, the highest microbial diversity was found in AD children. Based on 16S-rRNA and culture-dependent data, <i>Faecalibacterium</i> and <i>Ruminococcus</i> were present at the highest level in fecal samples of PDD-NOS and HC children. <i>Caloramator</i>, <i>Sarcina</i> and <i>Clostridium</i> genera were the highest in AD children. Compared to HC, the composition of Lachnospiraceae family also differed in PDD-NOS and, especially, AD children. Except for <i>Eubacterium siraeum</i>, the lowest level of Eubacteriaceae was found on fecal samples of AD children. The level of Bacteroidetes genera and some <i>Alistipes</i> and <i>Akkermansia</i> species were almost the highest in PDD-NOS or AD children as well as almost all the identified Sutterellaceae and Enterobacteriaceae were the highest in AD. Compared to HC children, <i>Bifidobacterium</i> species decreased in AD. As shown by Canonical Discriminant Analysis of Principal Coordinates, the levels of free amino acids and volatile organic compounds of fecal samples were markedly affected in PDD-NOS and, especially, AD children. If the gut microbiota differences among AD and PDD-NOS and HC children are one of the concomitant causes or the consequence of autism, they may have implications regarding specific diagnostic test, and/or for treatment and prevention.</p> </div

Fecal levels of free amino acids (FAA) in children.

<p>Concentration (mg/kg) of FAA found in fecal samples of Pervasive Developmental Disorder Not Otherwise Specified (PDD-NOS), autistic (AD) and healthy (HC) children. Data are the means of three independent experiments and standard deviations, performed in duplicate (n=6).</p

Total and active bacteria found in feces of children.

<p>Relative abundance (%) of total bacterial composition (16S-rDNA) (A) and metabolic active bacteria (16S-rRNA) (B) at the phylum level found in the fecal samples of Pervasive Developmental Disorder Not Otherwise Specified (PDD-NOS), autistic (AD) and healthy (HC) children. </p