In the present study, two lactobacilli strains, Lactobacillus rhamnosus H25 and Lactobacillus paracasei N24, used as adjunct cultures, were evaluated for their heat resistance both with and without ...prior heat adaptation and for their survival, at industrial scale, during the production and ripening of the Pecorino Siciliano cheese. In addition, the viability and persistence of the lactobacilli strains after passage through the gastrointestinal tract of healthy volunteers were evaluated by using rep-PCR analysis of viable cells. Both strains exhibited good heat resistance and survival throughout cheese production and ripening, and positively influenced the physico-chemical, the microbiological and the sensorial characteristics of the final product. In addition, the molecular typing of the lactobacilli isolates, retrieved from fecal samples of healthy volunteers during and after 15 days of the experimental cheese administration, revealed a high survival of the strains, highlighting their persistence during passage into the GI tract. In conclusion, this study proposes the two adjunct cultures as potential probiotic candidate deliverable by cheese.
•Pecorino ripened cheese was made at industrial scale using two adjunct cultures.•The adjunct strains were detected through the ripening till 180days.•The strains survived through the passage on the GI tract of healthy volunteers.•The potential probiotic strains survived also after stopping cheese administration.•The adjunct strains could be proposed as new probiotics for dairy use.
Administration of probiotics has been linked to immune regulation and changes in gut microbiota composition, with effects on atopic dermatitis (AD). In this study, we investigated amelioration of the ...symptoms of AD using Lactobacillus paracasei KBL382 isolated from the feces of healthy Koreans. Mice with Dermatophagoides farinae extract (DFE)-induced AD were fed 1 × 10
9
CFU d
−1
of L. paracasei KBL382 for 4 weeks. Oral administration of L. paracasei KBL382 significantly reduced AD-associated skin lesions, epidermal thickening, serum levels of immunoglobulin E, and immune cell infiltration. L. paracasei KBL382-treated mice showed decreased production of T helper (Th)1-, Th2-, and Th17-type cytokines, including thymic stromal lymphopoietin, thymus, and activation-regulated chemokine, and macrophage-derived chemokine, and increased production of the anti-inflammatory cytokine IL-10 and transforming growth factor-β in skin tissue. Intake of L. paracasei KBL382 also increased the proportion of CD4+ CD25+ Foxp3+ regulatory T cells in mesenteric lymph nodes. In addition, administration of L. paracasei KBL382 dramatically changed the composition of gut microbiota in AD mice. Administration of KBL382 significantly ameliorates AD-like symptoms by regulating the immune response and altering the composition of gut microbiota.
The aim of this work was to evaluate whether, in case of freeze-dried bacteria, the protective effect of a surrounding glassy matrix can be ascribed to its inherent restricted molecular mobility. ...Lactobacillus paracasei ssp. paracasei together with three different protectants (lactose, trehalose and dextran) was freeze-dried and stored at different temperatures and water activities (a sub(w)). The spin-spin relaxation time T sub(2) was determined by means of low resolution super(1)H-NMR spectroscopy and described in relation to the storage conditions and the glass transition temperature T sub(g). Compared to the disaccharides, dextran generally showed lower absolute T sub(2) values and a weaker dependence on storage a sub(w) and temperature. For lactose and trehalose, the plasticising effect of water and temperature was significantly stronger. Their relaxation time T sub(2) was shown to be only dependent on Delta T, the temperature distance to T sub(g). Furthermore, both disaccharides showed an increase in T sub(2) already 20-40 degree C below T sub(g). Thus, T sub(g) in reference to T sub(2) does not act as an absolute threshold. This fact could explain the finding of several publications that an absolute stability for freeze-dried bacteria can only be achieved 30-50 degree C below T sub(g). Further comparison between T sub(2) and the corresponding inactivation rates revealed that the relevance of mobility for the stability of lyophilisates is strongly temperature and system dependent. At low storage temperatures, a potential rate-limiting effect due to the lack of detrimental chemical and physical reactions is invisible. However, with increasing storage temperature, the restricted mobility is shown to become a rate limiting bottleneck. Thus, the harsher the environmental condition, the more relevant is the protective effect of a surrounding glassy matrix.
We previously identified a Neisseria flavescens strain in the duodenum of celiac disease (CD) patients that induced immune inflammation in ex vivo duodenal mucosal explants and in CaCo‐2 cells. We ...also found that vesicular trafficking was delayed after the CD‐immunogenic P31‐43 gliadin peptide‐entered CaCo‐2 cells and that Lactobacillus paracasei CBA L74 (L. paracasei‐CBA) supernatant reduced peptide entry. In this study, we evaluated if metabolism and trafficking was altered in CD‐N. flavescens‐infected CaCo‐2 cells and if any alteration could be mitigated by pretreating cells with L. paracasei‐CBA supernatant, despite the presence of P31‐43. We measured CaCo‐2 bioenergetics by an extracellular flux analyser, N. flavescens and P31‐43 intracellular trafficking by immunofluorescence, cellular stress by TBARS assay, and ATP by bioluminescence. We found that CD‐N. flavescens colocalised more than control N. flavescens with early endocytic vesicles and more escaped autophagy thereby surviving longer in infected cells. P31‐43 increased colocalisation of N. flavescens with early vesicles. Mitochondrial respiration was lower (P < .05) in CD‐N. flavescens‐infected cells versus not‐treated CaCo‐2 cells, whereas pretreatment with L. paracasei‐CBA reduced CD‐N. flavescens viability and improved cell bioenergetics and trafficking. In conclusion, CD‐N. flavescens induces metabolic imbalance in CaCo‐2 cells, and the L. paracasei‐CBA probiotic could be used to correct CD‐associated dysbiosis.
Long ripened cheeses, such as Grana Padano (GP), a Protected Designation of Origin (PDO) Italian cheese, harbor a viable microbiota mainly composed of non-starter lactic acid bacteria (NSLAB), which ...contribute to the final characteristics of cheese. The NSLAB species Lactobacillus rhamnosus, Lb. casei and Lb. paracasei are frequently found in GP, and form a closely related taxonomic group (Lb. casei group), making it difficult to distinguish the three species through 16S rRNA sequencing. SpxB, a metabolic gene coding for pyruvate oxidase in Lb. casei group, was recently used to distinguish the species within this bacterial group, both in pure cultures and in cheese, where it could provide an alternative energy source through the conversion of pyruvate to acetate. The aim of this work was to study the evolution of the metabolically active microbiota during different stages of GP ripening, targeting 16S rRNA to describe the whole microbiota composition, and spxB gene to monitor the biodiversity within the Lb. casei group. Furthermore, activation of pyruvate oxidase pathway was measured directly in cheese by reverse transcription real time PCR (RT–qPCR). The results showed that Lb. casei group dominates throughout the ripening and high-throughput sequencing of spxB allowed to identify four clusters inside the Lb. casei group. The dynamics of the sequence types forming the clusters were followed during ripening. Pyruvate oxidase pathway was expressed in cheese, showing a decreasing trend over ripening time. This work highlights how the composition of the microbiota in the early manufacturing stages influences the microbial dynamics throughout ripening, and how targeting of a metabolic gene can provide an insight into the activity of strains relevant for dairy products.
•16S rRNA HTS was used to monitor the microbiota of hard cooked Grana Padano cheese.•spxB HTS allows to study the dynamics of metabolically active Lb. casei group.•spxB sequence types follow different dynamics in the cheese-making.•spxB gene is expressed in cheese and decreases during ripening.•Microbiota of early fermentation stages influence population dynamics during ageing.
The purpose of this study was to investigate whether vaginal administration of probiotic Lactobacillus results in their colonization and persistence in the vagina and whether it promotes ...normalization and maintenance of pH and Nugent score. A single-arm, open-label controlled towards the baseline (pre-post) study including 35 apparently healthy women was conducted. Each woman was examined three times during the study. Women were instructed to receive daily for 7 days, the probiotic suppositories SYNBIO super( registered ) gin (Lactobacillus rhamnosus IMC 501 super( registered ) and Lactobacillus paracasei IMC 502 super( registered )). Vaginal swabs were collected during visit 1, 2, and 3 to determine the total lactobacilli count, the presence of the two administered bacteria, the measure of the pH, and the estimation of Nugent score. Evaluation of treatment tolerability was based on analysis of the type and occurrence of adverse events. The probiotic vaginal suppository was well tolerated and no side effects were reported. Intermediate Nugent score was registered in 40 % of women at visit 1 and these intermediate scores reverted to normal at day 7 (end of treatment) in 20 % of subjects. Administration of SYNBIO super( registered ) gin contributed to a significant increase in the lactobacilli level at visit 2. Molecular typing revealed the presence of the two strains originating from SYNBIO super( registered ) gin in 100 % of women at visit 2 and 34 % at visit 3. No significant changes were registered for pH between visits. The SYNBIO super( registered ) gin product is safe for daily use in healthy women and it could be useful to restore and maintain a normal vaginal microbiota.
•Novel, potentially probiotic, Lactobacillus paracasei K5 used as sourdough starter.•The K5 breads had good quality (rising, acidity, organic acids, consumer preference).•The K5 breads had more ...complex volatile profiles as shown by SPME-GC/MS.•The appearance of rope spoilage was delayed (macroscopic and PCR-DGGE analysis).•L. paracasei K5 can be used for good quality & extended shelf-life sourdough bread.
The novel Lactobacillus paracasei K5 strain, recently isolated from Greek cheese, was evaluated as potential sourdough bread starter. Breads were made using different amounts of L. paracasei sourdoughs as well as traditional sourdough for comparison. Quality characteristics of the breads (acidity and rising) were examined, as well as rope spoilage through macroscopic observations and molecular analysis (PCR-DGGE). The highest acidity levels (3.15 g lactic acid and 1.13 g acetic acid per kg of bread) and better resistance to rope spoilage were observed when bread contained 30% w/w L. paracasei K5 sourdough. Spoilage in the L. paracasei K5 breads was observed at 15–16 days, 5 days later than the control breads. In addition, L. paracasei K5 sourdough improved the bread sensory properties, as reflected by consumer preference and GC/MS analysis of aroma volatiles. Therefore, L. paracasei K5 can be successfully used for sourdough bread making with good quality and extended shelf-life.
Coeliac disease is an increasingly recognised pathology, induced by the ingestion of gluten in genetically predisposed patients. Undigested gliadin peptide can induce adaptive and innate immune ...response that unleash the typical intestinal mucosal alterations. A growing attention is paid to alternative therapeutic approaches to the gluten-free diet: one of these approaches is the use of probiotics and/or postbiotics. We performed lactic fermentation of rice flour with and without pH control, using Lactobacillus paracasei CBA L74 as fermenting strain. We evaluated bacterial growth, lactic acid production during fermentation and gliadin peptide P31-43 entrance in CaCo-2 cells with and without pH control. When pH control was applied no differences were observed in terms of bacterial growth; on the contrary, lactic acid production was greater, as expected. Both samples could inhibit the P31-43 entrance in CaCo-2 cells but the effect was significantly greater for samples obtained when the pH control was applied.
Allergic disease is one of the most important and common health problems worldwide. We have previously demonstrated that a fig leaf-derived lactic acid bacterium
(
)
IJH-SONE68 produces a novel ...exopolysaccharide (EPS). Furthermore, we have shown that the EPS inhibits the catalytic activity of hyaluronidase (EC 3.2.1.36) promoting inflammatory reactions. To evaluate the anti-allergy and anti-inflammatory effects of the EPS, in the present study, we employed the picryl-chloride-induced delayed-type (type IV) allergy model mice, which is used to evaluate the contact dermatitis. Oral administration of the EPS was observed to reduce the ear swelling in the model mice. We also observed that the overexpression of ear interleukin-4 (T helper (Th) 2 cytokine) mRNA and the increase in serum immunoglobulin E (IgE) are repressed. However, the expression of interferon-γ (Th1 cytokine) was not accelerated in all of the allergen-challenged model mice. The improvement may be responsible for the Th2 downregulation rather than the Th1 upregulation. In addition, the symptom of immediate-type (type I) allergy model mice was improved by oral administration of the IJH-SONE68 cell (data not shown). We can conclude that the IJH-SONE68-derived EPS is useful to improve the type I and IV allergies including atopic dermatitis.
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•L. Paracasei inhibited NF-κB activation to suppress TNF-α and IL-6 production.•L. Paracasei induced the expression of A20, SOCS1, SOCS3, and IRAK3.•IRAK4 inhibitor suppressed the ...expression of negative regulators.•Antibody against TLR2 attenuated the expression of negative regulators.•L. paracasei restrained NF-κB signaling in TLR2-IRAK4-dependent manner.
The application of the probiotic lactobacillus is suggested in the treatment of some inflammatory diseases of intestines due to its potential ability to attenuate inflammation. However, the mechanism is not completely understood. In PBMCs, Lactobacillus paracasei (L. Paracasei) down-regulated the LPS-induced production of TNF-α and IL-6. Using a macrophage-like differentiated THP-1 cell line induced by PMA, we investigated the effect of L. paracasei on the production of pro-inflammatory cytokines by monocyte-macrophages. Treatment of the differentiated THP-1 cells with L. paracasei either concurrently with or before LPS challenge attenuated the LPS-induced secretion of TNF-α and IL-1β. This effect was due to a decrease in IκB phosphorylation and NF-κB nuclear translocation. Furthermore, treatment of the differentiated THP-1 cells with L. paracasei induced the expression of negative regulators of the NF-κB signaling pathway, including the deubiquitinating enzyme A20, suppressor of cytokine signaling (SOCS) 1, SOCS3, and IL-1 receptor-associated kinase (IRAK) 3. Pretreatment with an IRAK4 inhibitor suppressed the L. paracasei-induced expression of these negative regulators and further increased the LPS-mediated expressions of TNF-α and IL-1β. Moreover, treatment with an antibody against Toll-like receptor (TLR) 2 reversed the effect of L. paracasei on inducing negative regulators and inhibiting TNF-α and IL-1β productions. Our findings suggest that L. paracasei inhibits the production of pro-inflammatory cytokines by monocyte-macrophages via the induction of negative regulators of the NF-κB signaling pathway in a TLR2-IRAK4-dependent manner.
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