Activation of the toll-like receptor 2 signaling pathway by GAPDH from bacterial strain RD055328.

We characterized the membrane vesicle fraction (RD-MV fraction) from bacterial strain RD055328, which is related to members of the genus Companilactobacillus and Lactiplantibacillus plantarum. RD-MVs and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were detected in the RD-MV fraction. Immunoglobulin A (IgA) was produced by Peyer's patch cells following the addition of the RD-MV fraction. In the presence of the RD-MV fraction, RAW264 cells produced the pro-inflammatory cytokine IL-6. Recombinant GAPDH probably induced the production of IL-6 by RAW264 cells via superficial toll-like receptor 2 (TLR2) recognition. A confocal laser scanning microscopy image analysis indicated that RD-MVs and GAPDH were taken up by RAW264 cells. GAPDH wrapped around RAW264 cells. We suggest that GAPDH from strain RD055328 enhanced the production of IgA by acquired immune cells via the production of IL-6 by innate immune cells through TLR2 signal transduction.

Antagonistic activity of lactic acid bacteria isolated from Minas artisanal cheeses against Brucella abortus / Atividade antagonista de bactérias lácticas isoladas de queijos Minas artesanais contra Brucella abortus

This study aimed to evaluate methods for studying the in vitro antimicrobial activity of lactic acid bacteria (LAB) against Brucella abortus and to evaluate the antagonistic effect of LAB on the viability of this pathogen. A total of 18 LAB strains (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; and Lactobacillus brevis,n = 2), isolated from Minas artisanal cheeses produced in three regions (Canastra, Campos das Vertentes, and Araxá) of Minas Gerais State, Brazil, were tested for their antimicrobial activity against B. abortus using three methods: spot-on-lawn, agar well diffusion assay, and antagonistic activity of the culture supernatants. None of the tested LAB strains could inhibit B. abortus in the spot-on-lawn and agar-well diffusion assays. The supernatants produced by LAB had an acidic pH, with intensity depending on bacterial growth and strain, and could inhibit the growth of B. abortus. In contrast, pH-neutralized (pH 7.0) LAB supernatants did not suppress the growth of B. abortus. The results showed that the best technique to study the in vitro antagonism of LAB against B. abortus was the antagonistic activity of culture supernatants. The growth of B. abortus may have been inhibited by acid production.(AU)

Este estudo teve como objetivo avaliar métodos de estudo in vitro da atividade antimicrobiana de bactérias lácticas contra Brucella abortus e avaliar o efeito antagônico das mesmas sobre a viabilidade deste patógeno. Um total de 18 amostras de bactérias lácteas (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; e Lactobacillus brevis, n = 2), isoladas de exemplares de Queijo Minas Artesanal produzidos em três regiões (Canastra, Campos das Vertentes e Araxá) do estado de Minas Gerais, Brasil, foram testados quanto à sua atividade antimicrobiana contra B. abortus usando três métodos: spot-on-lawn, ensaio de difusão em poço e atividade antagonista de sobrenadante de cultura. Nenhuma das cepas testadas foi capaz de inibir B. abortus nos ensaios spot-on-lawm e de difusão em poço. Os sobrenadantes produzidos pelas bactérias lácteas apresentaram pH ácido, com intensidade dependente do crescimento bacteriano e da amostra, podendo inibir o crescimento de B. abortus. Em contraste, os sobrenadantes com pH neutralizado (pH 7,0) não inibiram o crescimento de B. abortus. Os resultados mostraram que a melhor técnica para estudar o antagonismo in vitro de bactérias lácteas contra B. abortus foi a atividade antagonista de sobrenadante de cultura. O crescimento de B. abortus pode ter sido inibido pela produção de ácido.(AU)

Multifunctional Probiotic and Functional Properties of Lactiplantibacillus plantarum LRCC5314, Isolated from Kimchi.

In this study, the survival capacity (acid and bile salt tolerance, and adhesion to gut epithelial cells) and probiotic properties (enzyme activity-inhibition and anti-inflammatory activities, inhibition of adipogenesis, and stress hormone level reduction) of Lactiplantibacillus plantarum LRCC5314, isolated from kimchi (Korean traditional fermented cabbage), were investigated. LRCC5314 exhibited very stable survival at ph 2.0 and in 0.2% bile acid with 89.9% adhesion to Caco-2 intestinal epithelial cells after treatment for 2 h. LRCC5314 also inhibited the activities of α-amylase and α-glucosidase, which are involved in elevating postprandial blood glucose levels, by approximately 72.9% and 51.2%, respectively. Treatment of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells with the LRCC5314 lysate decreased the levels of the inflammatory factors nitric oxide, tumor necrosis factor (TNF-α), interleukin (IL)-1ß, and interferon-γ by 88.5%, 49.3%, 97.2%, and 99.8%, respectively, relative to those of the cells treated with LPS alone. LRCC5314 also inhibited adipogenesis in differentiating preadipocytes (3T3-L1 cells), showing a 14.7% decrease in lipid droplet levels and a 74.0% decrease in triglyceride levels, as well as distinct reductions in the mRNA expression levels of adiponectin, FAS, PPAR/γ, C/EBPα, TNF-α, and IL-6. Moreover, LRCC5314 reduced the level of cortisol, a hormone with important effect on stress, by approximately 35.6% in H295R cells. L. plantarum LRCC5314 is identified as a new probiotic with excellent in vitro multifunctional properties. Subsequent in vivo studies may further demonstrate its potential as a functional food or pharmabiotic.

Lentilactobacillus fungorum sp. nov., isolated from spent mushroom substrates.

In Japan, during a screening of lactic acid bacteria in spent mushroom substrates, an unknown bacterium was isolated and could not be assigned to any known species. Strain YK48GT is Gram-stain-positive, rod-shaped, non-motile, non-spore-forming and catalase-negative. The isolate grew in 0-4 % (w/v) NaCl, at 15-37 °C (optimum, 30 °C) and at pH 4.0-8.0 (optimum, pH 6.0). The genomic DNA G+C content of strain YK48GT was 42.5 mol%. Based on its 16S rRNA gene sequence, strain YK48GT represented a member of the genus Lentilactobacillus and showed the highest pairwise similarity to Lentilactobacillus rapi DSM 19907T (97.86 %). Phylogenetic analyses based on amino acid sequences of 466 shared protein-encoding genes also revealed that the strain was phylogenetically positioned in the genus Lentilactobacillus but did not suggest an affiliation with previously described species. The average nucleotide identity and digital DNA-DNA hybridization values between strain YK48GT and the type strains of phylogenetically related species were 72.2-76.6% and 19.0-21.2 %, respectively, indicating that strain YK48GT represents a novel species within the genus Lentilactobacillus. Phenotypic data further confirmed the differentiation of strain YK48GT from other members of the genus Lentilactobacillus. According to the results of the polyphasic characterization presented in this study, strain YK48GT represents a novel species of the genus Lentilactobacillus, for which the name Lentilactobacillus fungorum sp. nov. is proposed. The type strain is YK48GT (=JCM 32598T=DSM 107968T).

Lentilactobacillus kosonis sp. nov., isolated from kôso, a Japanese sugar-vegetable fermented beverage.

A novel lactic acid-producing, Gram-stain-positive, catalase-negative and rod-shaped strain, designated as strain C06_No.73T, was isolated from a traditional Japanese fermented beverage called kôso. According to the results of phylogenetic analysis based on 16S rRNA gene sequences, strain C06_No.73T belongs to the genus Lentilactobacillus. The closest type strain was Lentilactobacillus curieae CCTCC M 2011381T, with a sequence identity of 98.1 %. The identity values with other strains were all below 97 %. The isolate propagated under the conditions of 18-39 °C (optimum, 27 °C for 48 h incubation) and pH 4.0-7.0 (optimum, pH 6.5). The G+C content of its genomic DNA was determined to be 37.9 mol%. The main fatty acids were C16 : 0, C18 : 1 ω7c, C18 : 1 ω9c and C19 : 0 cyclopropane 11,12. The major polar lipid was identified as phosphatidylglycerol. No isoprenoid quinone was detected. The predominant cell-wall amino acids were lysine, alanine, glutamic acid and aspartic acid. Neither meso-diaminopimelic acid nor ornithine were detected. On the basis of this polyphasic taxonomic study, the isolate is concluded to represent a novel species, for which the name Lentilactobacillus kosonis sp. nov. is proposed. The type strain is C06_No.73T (=NBRC 111893T=BCRC 81282T).

Apilactobacillus nanyangensis sp. nov., Secundilactobacillus hailunensis sp. nov., Secundilactobacillus yichangensis sp. nov., Levilactobacillus andaensis sp. nov., Levilactobacillus wangkuiensis sp. nov., Levilactobacillus lanxiensis sp. nov., Lacticaseibacillus mingshuiensis sp. nov. and Lacticaseibacillus suilingensis sp. nov., isolated from traditional Chinese pickle and the gut of honeybee (Apis mellifera).

Thirteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and the gut of honeybee (Apis mellifera). These strains were characterized using a polyphasic taxonomic approach. The data demonstrated that 12 of the 13 strains represented eight novel species belonging to the genera Apilactobacillus, Secundilactobacillus, Levilactobacillus and Lacticaseibacillus; strains HN36-1T, 887-11T, F79-211-2T, 866-3T, 6-5(1)T, 13B17T, 117-1T and ZW152T were designated as the type strains. Based upon the data of polyphasic characterization obtained in the present study, eight novel species, Apilactobacillus nanyangensis sp. nov., Secundilactobacillus hailunensis sp. nov., Secundilactobacillus yichangensis sp. nov., Levilactobacillus andaensis sp. nov., Levilactobacillus wangkuiensis sp. nov., Levilactobacillus lanxiensis sp. nov., Lacticaseibacillus mingshuiensis sp. nov. and Lacticaseibacillus suilingensis sp. nov., are proposed and the type strains are HN36-1T (=JCM 33867T=CCTCC AB 2019385T), 887-11T (=NCIMB 15201T=CCM 8950T=JCM 33864T=CCTCC AB 2018396T), F79-211-2T (=NCIMB 15254T=JCM 33866T=CCTCC AB 2019384T), 866-3T (=JCM 33863T=CCTCC AB 2019383T), 6-5(1)T (=NCIMB 15229T=CCM 8977T=JCM 33564T=CCTCC AB 2019168T), 13B17T (=NCIMB 15230T=CCM 8979T=JCM 33565T=CCTCC AB 2019167T), 117-1T (=NCIMB 15232T=CCM 8980T=JCM 33567T) and ZW152T (=JCM 34363T=CCTCC AB 2020299T=LMG 32143T=CCM 9110T), respectively.

Secundilactobacillus folii sp. nov., isolated from fermented tea leaves in Thailand.

A Gram-stain-positive, catalase-negative, rod-shaped, non-motile, non-spore-forming, and facultatively anaerobic strain CRM56-3T, isolated from fermented tea leaves collected from Chiang Rai province, Thailand, was characterized based on a polyphasic approach. The strain produced dl-lactic acid heterofermentatively from glucose. It grew at 15-42 °C (optimum at 30 °C), pH 3.5-8.0 (optimum pH 6.0) and in 1-4 % (w/v) NaCl. Strain CRM56-3T contained C16:0, C19:0 cyclo ω8c, and C18:1 ω7c, and/or C18:1 ω6c as major cellular fatty acids. Based on 16S rRNA gene sequence analysis, strain CRM56-3T belongs to the genus Secundilactobacillus and was closely related to Secundilactobacillus odoratitofui DSM 19909T (99.2 %), S. collinoides JCM 1123T (98.9 %), and S. paracollinoides DSM 15502T (98.7 %). The draft genome of strain CRM56-3T contained 2681617 bp with 2413 coding sequences and DNA G+C content determined from genome sequence of 44.5 mol%. The digital DNA-DNA hybridization (dDDH) between strain CRM56-3T and S. odoratitofui DSM 19909T, S. collinoides JCM 1123T, and S. paracollinoides DSM 15502T were 19.5, 20.4, and 21.6 %, respectively. The average nucleotide identity (ANIm) and the average amino acid identity (AAI) between strain CRM56-3T and closely related strains were lower than 85.0 and 80.0 %, respectively. The strain CRM56-3T was clearly distinguished from related Secundilactobacillus species by its phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequence similarity, and the draft genome analysis. Therefore, the strain represents a novel species of the genus Secundilactobacillus, for which the name of Secundilactobacillus folii sp. nov. is proposed. The type strain is CRM56-3T (=JCM 34223T=LMG 31663T=TISTR 2851T).

Behaviour of citrus pectin and modified citrus pectin in an azoxymethane/dextran sodium sulfate (AOM/DSS)-induced rat colorectal carcinogenesis model.

Large intestine cancer is one of the most relevant chronic diseases taking place at present. Despite therapies have evolved very positively, this pathology is still under deep investigation. One of the recent approaches is the prevention by natural compounds such as pectin. In this paper, we have assessed the impact of citrus pectin and modified citrus pectin on colorectal cancer in rats (Rattus norvegicus F344) to which azoxymethane and DSS were supplied. The lowest intake of food and body weight were detected in animals fed with citrus pectin, together with an increase in the caecum weight, probably due to the viscosity, water retention capacity and bulking properties of pectin. The most striking feature was that, neither citrus pectin nor modified citrus pectin gave rise to a tumorigenesis prevention. Moreover, in both, more than 50% of rats with cancer died, probably ascribed to a severe dysbiosis state in the gut, as shown by the metabolism and metagenomics studies carried out. This was related to a decrease of pH in caecum lumen and increase in acetate and lactic acid levels together with the absence of propionic and butyric acids. A relevant increase in Proteobacteria (Enterobacteriaceae) were thought to be one of the reasons for enteric infection that could have provoked the death of rats and the lack of cancer prevention. However, a reduction of blood glucose and triacylglycerides level and an increase of Bifidobacterium and Lactobacillaceae were found in animals that intake pectin, as compared to universal and modified citrus pectin feeding.

Effects of Olive and Pomegranate By-Products on Human Microbiota: A Study Using the SHIME® in Vitro Simulator.

Two by-products containing phenols and polysaccharides, a "pâté" (OP) from the extra virgin olive oil milling process and a decoction of pomegranate mesocarp (PM), were investigated for their effects on human microbiota using the SHIME® system. The ability of these products to modulate the microbial community was studied simulating a daily intake for nine days. Microbial functionality, investigated in terms of short chain fatty acids (SCFA) and NH4+, was stable during the treatment. A significant increase in Lactobacillaceae and Bifidobacteriaceae at nine days was induced by OP mainly in the proximal tract. Polyphenol metabolism indicated the formation of tyrosol from OP mainly in the distal tract, while urolithins C and A were produced from PM, identifying the human donor as a metabotype A. The results confirm the SHIME® system as a suitable in vitro tool to preliminarily investigate interactions between complex botanicals and human microbiota before undertaking more challenging human studies.

Specific gut microbiome members are associated with distinct immune markers in pediatric allogeneic hematopoietic stem cell transplantation.

BACKGROUND: Increasing evidence reveals the importance of the microbiome in health and disease and inseparable host-microbial dependencies. Host-microbe interactions are highly relevant in patients receiving allogeneic hematopoietic stem cell transplantation (HSCT), i.e., a replacement of the cellular components of the patients' immune system with that of a foreign donor. HSCT is employed as curative immunotherapy for a number of non-malignant and malignant hematologic conditions, including cancers such as acute lymphoblastic leukemia. The procedure can be accompanied by severe side effects such as infections, acute graft-versus-host disease (aGvHD), and death. Here, we performed a longitudinal analysis of immunological markers, immune reconstitution and gut microbiota composition in relation to clinical outcomes in children undergoing HSCT. Such an analysis could reveal biomarkers, e.g., at the time point prior to HSCT, that in the future could be used to predict which patients are of high risk in relation to side effects and clinical outcomes and guide treatment strategies accordingly. RESULTS: In two multivariate analyses (sparse partial least squares regression and canonical correspondence analysis), we identified three consistent clusters: (1) high concentrations of the antimicrobial peptide human beta-defensin 2 (hBD2) prior to the transplantation in patients with high abundances of Lactobacillaceae, who later developed moderate or severe aGvHD and exhibited high mortality. (2) Rapid reconstitution of NK and B cells in patients with high abundances of obligate anaerobes such as Ruminococcaceae, who developed no or mild aGvHD and exhibited low mortality. (3) High inflammation, indicated by high levels of C-reactive protein, in patients with high abundances of facultative anaerobic bacteria such as Enterobacteriaceae. Furthermore, we observed that antibiotic treatment influenced the bacterial community state. CONCLUSIONS: We identify multivariate associations between specific microbial taxa, host immune markers, immune cell reconstitution, and clinical outcomes in relation to HSCT. Our findings encourage further investigations into establishing longitudinal surveillance of the intestinal microbiome and relevant immune markers, such as hBD2, in HSCT patients. Profiling of the microbiome may prove useful as a prognostic tool that could help identify patients at risk of poor immune reconstitution and adverse outcomes, such as aGvHD and death, upon HSCT, providing actionable information in guiding precision medicine.

Effect of nutritional interventions with quercetin, oat hulls, ß-glucans, lysozyme and fish oil on performance and health status related parameters of broilers chickens.

1. An experiment was conducted to evaluate the effects of technical feed ingredients between 14 and 28 d of age on performance and health status of broilers (d 14-35) fed diets with a high inclusion rate of rapeseed meal as a nutritional challenge. It was hypothesized that the feed ingredients would improve health status related parameters. 2. A total of 1008 one-day-old male Ross 308 chicks were distributed over 36 floor pens and allocated to one of six iso-caloric (AMEN 13 MJ/kg) growing diets (d 15-28): a control and five test diets supplemented with quercetin (400 mg/kg), oat hulls (50 g/kg), ß-glucan (100 mg/kg), lysozyme (40 mg/kg) or fish oil ω-3 fatty acids (40 g/kg), with six replicate pens per treatment. 3. Dietary inclusion of oat hulls and lysozyme resulted in a reduction in broiler performance during the first week after providing the experimental diets. 4. No effect of interventions on the microbiota diversity in the jejunum and ileum was observed. Ileal microbiota composition of birds fed oat hulls differed from the other groups, as shown by a higher abundance of the genus Enterococcus, mainly at the expense of the genus Lactobacillus. 5. In the jejunum, villus height and crypt depth of lysozyme-fed birds at d 28 were decreased compared to the control group. Higher total surface area of villi occupied by goblet cells and total villi surface area in jejunum (d 21 and 28) were observed in chickens fed oat hulls compared to other groups. 6. Genes related to the growth-factor-activity pathway were more highly expressed in birds fed ß-glucan compared to the control group, while the genes related to anion-transmembrane-transporter-activity pathway in the quercetin- and oat hull-fed birds were less expressed. The genes differently expressed between dietary interventions did not seem to be directly involved in immune related processes. 7. It was concluded that the tested nutritional interventions in the current experiment only marginally effected health status related parameters.

Polyphasic approach to study physico-chemical, microbiological and sensorial characteristics of artisanal Nicastrese goat's cheese.

Nicastrese goat's cheese is produced in the South of Italy under traditional procedures, from raw goat milk without any starter cultures addition. Samples from milk to ripened cheese provided by 4 different farms were subjected to a polyphasic approach to study their physico-chemical, microbiological and sensorial characteristics. In addition, volatile organic compounds formation in the final products was studied. Overall, gross composition and microbiological data revealed a significant variability among samples, which was confirmed by both the volatile organic compounds generated in the final products and by the sensorial data. Conventional technique allowed us to identify 720 isolates, mainly belonging to Lactococcus lactis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus brevis, Leuconostoc mesenteroides, and Enterococcus faecalis. Culture-independent methods revealed shifts in the microbial community structure, with an increase in biodiversity of metabolically active bacterial species, from milk to cheese samples. Analysis of volatile organic compounds (VOCs) allowed the identification of 36 compounds; free fatty acids and ketones represented the main detected, followed by alcohols and esters. Moreover, statistical analysis was performed in order to correlate VOCs to bacterial species. Data showed that ester compounds as well as alcohol and aldehydes were positively correlated to NSLAB, indicating that the occurrence of L. casei, L. plantarum and L. brevis species is relevant for the VOCs formation in the final product.

Polyphasic Microbial Analysis of Traditional Korean Jeung-Pyun Sourdough Fermented with Makgeolli.

Jeung-pyun, a fermented rice cake, is prepared by fermenting rice sourdough using makgeolli, a traditional Korean rice wine, in the presence of yeast and lactic acid bacteria (LAB). The goal of this study was to conduct biochemical and microbial analyses of five different rice sourdoughs, each fermented with a different commercial makgeolli, using culture-dependent and culture-independent approaches. All sourdough samples fermented with different makgeolli for 6.5 h showed different profiles in pH, total titratable acidity, organic acid concentration, and microbial growth. LAB belonging to different genera were identified based on colony morphology on modified MRS and sourdough bacteria agar medium. PCR-denaturinggradient gel electrophoresis analyses of the five sourdoughs showed different bands corresponding to LAB and yeast. 16S/26S rRNA gene sequence analyses of the samples confirmed that the predominant LAB in the five fermented rice doughs was Lactobacillus plantarum, Lb. pentosus, and Lb. brevis. Various other Lactobacillus spp. and Saccharomyces cerevisiae were common in all five fermented samples. This study provides comprehensive and comparative information on the microflora involved in fermentation of rice sourdough and signifies the need to develop effective starters to enrich the quality of jeung-pyun.

Biocide tolerance, phenotypic and molecular response of lactic acid bacteria isolated from naturally-fermented Aloreña table to different physico-chemical stresses.

Lactic acid bacteria (LAB) isolated throughout the fermentation process of Aloreña table olives were found to be resistant at least to three antibiotics (Casado Muñoz et al., 2014); however, most were sensitive to the biocides tested in this study (with minimum inhibitory concentrations [MIC] below the epidemiological cut-off values). 2-15% of the isolates were found to be biocide resistant: Leuconostoc Pseudomesenteroides, which were resistant to hexachlorophene, and Lactobacillus pentosus to cetrimide and hexadecylpiridinium. We analyzed the effect of different physico-chemical stresses, including antimicrobials, on the phenotypic and genotypic responses of LAB, providing new insights on how they become resistant in a changing environment. Results indicated that similar phenotypic responses were obtained under three stress conditions: antimicrobials, chemicals and UV light. Susceptibility patterns to antibiotics changed: increasing MICs for ampicillin, chloramphenicol, ciprofloxacin, teicoplanin and tetracycline, and decreasing the MICs for clindamycin, erythromycin, streptomycin and trimethoprim in most strains. Statistically, cross resistance between different antibiotics was detected in all stress conditions. However, expression profiles of selected genes involved in stress/resistance response (rpsL, recA, uvrB and srtA) differed depending on the stress parameter, LAB species and strain, and the target gene. We conclude that, despite the uniform phenotypic response to stresses, the repertoire of induced and repressed genes differs. So, a search for a target to improve stress tolerance of LAB, especially those of importance as starter/protective cultures or probiotics, may depend on the individual screening of each strain, even though we could predict the antibiotic phenotypic response to all stresses.

Molecular detection of intrauterine microbial colonization in women with endometriosis.

OBJECTIVE: Increased intrauterine microbial colonization by bacteria culture method and occurrence of endometritis have been reported in women with endometriosis. Here we investigated microbial colonization in intrauterine environment and cystic fluid of women with and without endometriosis by molecular approach. STUDY DESIGN: This is a case-controlled biological study with a total of 32 women each with and without endometriosis. Among them, 16 each in these two groups of women received treatment with gonadotropin-releasing hormone agonist (GnRHa). Pattern of microbial colonization in endometrial swabs and endometrioma/non-endometrioma cystic fluid was examined using broad-range polymerase-chain reaction (PCR) amplification of bacteria targeting 16S rRNA gene (rDNA). After quantification of index PCR product, 16S rDNA metagenome sequence analysis was done by Illumina Miseq system. RESULTS: A wide proportion (0.01-97.8%) of multiple bacteria was detected in both endometrial swabs and cystic fluid collected from women with and without endometriosis. 16S metagenome assay indicated that proportion of Lactobacillacae was significantly decreased (p<0.01) and of Streptococcaceae, Staphylococaceae, Enterobacteriaceae was significantly increased (p<0.05 for each) in GnRHa-treated women with endometriosis than in GnRHa-untreated women. While bacteria culture method failed to detect a single colony, 16S metagenome assay could detect significantly higher percentage of Streptococcaceae (p<0.01) and Staphylococaceae (p<0.05) in the cystic fluid derived from women with ovarian endometrioma comparing to that in cystic fluid collected from non-endometrioma cysts. CONCLUSION: These findings indicate the occurrence of sub-clinical infection in intrauterine environment and in the cystic fluid of ovarian endometrioma. Additional side effect of GnRHa treatment in promoting silent intrauterine and/or ovarian infection should be considered.

Transfer, composition and technological characterization of the lactic acid bacterial populations of the wooden vats used to produce traditional stretched cheeses.

The biofilms of 12 wooden vats used for the production of the traditional stretched cheeses Caciocavallo Palermitano and PDO Vastedda della valle del Belìce were investigated. Salmonella spp. and Listeria monocytogenes were never detected. Total coliforms were at low numbers with Escherichia coli found only in three vats. Coagulase-positive staphylococci (CPS) were below the enumeration limit, whereas lactic acid bacteria (LAB) dominated the surfaces of all vats. In general, the dominance was showed by coccus LAB. Enterococci were estimated at high numbers, but usually between 1 and 2 Log cycles lower than other LAB. LAB populations were investigated at species and strain level and for their technological properties relevant in cheese production. Eighty-five strains were analysed by a polyphasic genetic approach and allotted into 16 species within the genera Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Streptococcus. Enterococcus faecium was found in all wooden vats and the species most frequently isolated were Enterococcus faecalis, Lactococcus lactis, Leuconostoc mesenteroides, Pediococcus acidilactici and Streptococcus thermophilus. The study of the quantitative data on acidification rate, autolysis kinetics, diacetyl production, antibacterial compound generation and proteolysis by cluster and principal component analysis led to the identification of some strains with promising dairy characteristics. Interestingly, a consistent percentage of LAB was bacteriocin-like inhibitory substances (BLIS) producer. Thus, the microbial biofilms of the wooden vats analysed in this study might contribute actively to the stability of the final cheeses.

Whole grain oats improve insulin sensitivity and plasma cholesterol profile and modify gut microbiota composition in C57BL/6J mice.

BACKGROUND: Whole grain consumption reduces the risk of major chronic diseases. It is not clear how whole grains exert their beneficial effects. OBJECTIVE: The aim was to compare the physiologic effects of whole grain oat (WGO) flour with low bran oat (LBO) flour. METHODS: Two AIN-93G-based diets were formulated with either WGO or LBO flour. Five-week-old male C57BL/6J mice were fed LBO (n = 11) and WGO (n = 13) diets for 8 wk. Cecal microbiota was profiled by pyrosequencing of the 16S ribosomal RNA gene. Data are reported as means ± SEMs or antilogs of the mean (mean - SEM, mean + SEM). RESULTS: The weight gain was 14.6% less in the WGO group during week 7 (P = 0.04). WGO improved insulin sensitivity as reflected by significantly lower plasma insulin [1500 (1370, 1650) ng/L vs. 2340 (2090, 2620) ng/L; P = 0.006], C-peptide (3980 ± 548 ng/L vs. 7340 ± 1050 ng/L; P = 0.007), and homeostasis model assessment-estimated insulin resistance (21.4 ± 2.3 vs. 34.7 ± 4.9; P = 0.03). Plasma total cholesterol was 9.9% less and non-HDL cholesterol was 11% less in the WGO group. A comparison of relative abundance indicated Prevotellaceae, Lactobacillaceae, and Alcaligenaceae families were 175.5% (P = 0.03), 184.5% (P = 0.01), and 150.0% (P = 0.004), respectively, greater in the WGO group and Clostridiaceae and Lachnospiraceae families were 527% (P = 0.004) and 62.6% (P = 0.01), respectively, greater in the LBO group. Cecal microbiota composition predicts 63.9% variation in plasma insulin and 88.9% variation in plasma non-HDL cholesterol. CONCLUSIONS: In mice, WGOs improved insulin sensitivity and plasma cholesterol profile compared with LBOs, and the effects were associated with the changes in cecal microbiota composition. Increasing WGO consumption may help improve insulin sensitivity and dyslipidemia in chronic diseases.

Volatile compounds and bacterial community dynamics of chestnut-flour-based sourdoughs.

The aims of this study were the monitoring of the microbial dynamics by means of a polyphasic approach based on conventional isolation techniques and PCR-DGGE-based methods in different chestnut-based sourdoughs and the evaluation of the impact of fermentation on volatile organic compounds formation during sourdoughs ripening. Members of the Lactobacillus plantarum group and Pediococcus pentosaceous dominated the sourdough ecosystems. Nevertheless, RAPD-PCR allowed recording a relevant genotypic biodiversity among strains coming from gluten-free flour combinations. Volatile compounds were characterised by GC/MS. A total of 59 volatile compounds were identified, mainly alcohols, esters, acids, aldehydes and ketones. Principal component analysis of samples at the beginning and at the end of ripening offered a good separation of the samples and highlighted the effect of fermentation on the sensorial profile.