Genomic stability and phenotypic characteristics of industrially-produced Lacticaseibacillus rhamnosus GG in a yogurt matrix

2021 ◽  
Author(s):  
Marianne Stage ◽  
Yan Hui ◽  
Dennis Sandris Nielsen ◽  
Natalia Ivonne Vera-Jiménez ◽  
Jeanne Olsen ◽  
...  
Keyword(s):  
Starter Culture ◽  
Intestinal Barrier ◽  
Genomic Stability ◽  
Probiotic Strain ◽  
Intestinal Barrier Function ◽  
Probiotic Properties ◽  
Phenotypic Characteristics ◽  
Adhesion Properties ◽  
Acid And Bile Tolerance

Lacticaseibacillus rhamnosus GG is a widely marketed probiotic with well-documented probiotic properties. Previously, deletion of the mucus-adhesive spaCBA-srt C1 genes was reported in dairy isolates. Here, we examined the genome preservation of industrially-produced L. rhamnosus GG (DSM 33156) co-fermented in yogurts. In total, DNA of 66 samples, including 60 isolates, was sequenced. Population samples and 59 isolates exhibited an intact genome. One isolate exhibited loss of spaCBA-srt C1. In addition, we examined phenotypes related to the probiotic properties of L. rhamnosus GG either from frozen pellets or co-fermented in yogurt. L. rhamnosus GG from frozen pellets induced a response in intestinal barrier function in vitro , in contrast to frozen pellets of the starter culture. Yogurt matrix, containing only the starter culture, induced a response, but co-fermentation with L. rhamnosus GG induced a higher response. Conversely, only the starter culture stimulated cytokine secretion in dendritic cells, and it was observed that the addition of L. rhamnosus GG to the starter culture reduced the response. We conclude that the L. rhamnosus GG genome is preserved in yogurt and that common in vitro probiotic effects of L. rhamnosus GG are observed when examined in the yogurt matrix. Importance Lacticaseibacillus rhamnosus GG is a well-documented probiotic strain recognized for its high acid and bile tolerance and adhesion properties to enterocytes and mucus. The strain exhibits SpaCBA pili, which have been demonstrated to play an important role in adhesion and therefore are relevant for persistence in the gastrointestinal tract. Recently we demonstrated that the genome and phenotypes of L. rhamnosus GG are preserved throughout an industrial production pipeline. However, as gene deletions in L. rhamnosus GG were previously reported in isolates from dairy products, a key question on the genomic stability of L. rhamnosus GG in a yogurt matrix remained. The aim of this study was to analyze genome stability and phenotypic characteristics of L. rhamnosus GG in yogurt. We found that the genome of L. rhamnosus GG is well conserved when co-fermented in yogurt. Some phenotypic characteristics are consistent in all product matrixes, while other characteristics are modulated.

A Washed-Curd Goat's Cheese as a Vehicle for Delivery of a Potential Probiotic Bacterium: Lactobacillus delbrueckii subsp. lactis UO 004

2005 ◽  
Vol 68 (12) ◽  
pp. 2665-2671 ◽  
Author(s):  
M. FERNANDA FERNÁNDEZ ◽  
TERESA DELGADO ◽  
SOLEDAD BORIS ◽  
ANA RODRÍGUEZ ◽  
COVADONGA BARBÉS
Keyword(s):  
High Performance ◽  
Starter Culture ◽  
Food Product ◽  
Probiotic Strain ◽  
Probiotic Bacterium ◽  
Lactobacillus Delbrueckii ◽  
Probiotic Properties ◽  
Acid And Bile Tolerance ◽  
Probiotic Food ◽  
Lactobacillus Delbrueckii Subsp

This study characterizes the probiotic properties of Lactobacillus delbrueckii subsp. lactis UO 004 and examines its suitability for making cheese. This strain was isolated from infant feces and shows interesting features, such as acid and bile tolerance, adherence to intestinal epithelial cells, and inhibition of the growth of certain enteropathogens, that support its potential use as a probiotic strain. In this regard, the suitability of a washed-curd cheese (Vidiago type) made with goat's milk as a delivery system for this probiotic strain was assessed. Lactobacillus delbrueckii subsp. lactis UO 004 was incorporated into a starter culture (IPLA 001). Changes in the overall composition of control and experimental cheeses were determined during ripening through bacteriological, chemical, high-performance liquid chromatography, and gas chromatography analyses. Slight changes in the gross composition and appreciable differences in the flavor compounds profile were observed between control and experimental cheeses. This strain was capable of surviving at high cell numbers (108 to 109 CFU/g) in cheeses after 28 days of ripening without adversely affecting sensory criteria or appearance of the cheese, thus satisfying the criteria for a probiotic food product.

Challenges in translational research on probiotic lactobacilli: from in vitro assays to clinical trials

2013 ◽  
Vol 4 (1) ◽  
pp. 83-100 ◽  
Author(s):  
M. Meijerink ◽  
A. Mercenier ◽  
J.M. Wells
Keyword(s):  
Translational Research ◽  
Selection Criteria ◽  
Molecular Mechanisms ◽  
Current Knowledge ◽  
Intestinal Barrier ◽  
Probiotic Strain ◽  
In Vitro Assays ◽  
Intestinal Barrier Function

Beneficial effects of certain probiotic strains have been established in the treatment and prevention of various immune and intestinal disorders in humans, including allergic diseases, chronic inflammatory diseases and diarrhoea. The proposed mechanisms underlying the immunomodulatory effects of probiotics in humans are not understood in precise detail but include enhancement of intestinal barrier function, altered epithelial signalling, competition with pathogens and effects on immune cells and immunity depending on the probiotic strain. The publication of controversial or inconclusive probiotic studies in humans highlights the need for a better understanding of the mechanisms and improved strain selection criteria. This review focuses on the immunomodulatory properties of lactobacilli and bifidobacteria in vitro and in vivo, current knowledge concerning the mechanisms in vivo and challenges in translational research on probiotics. A better understanding of the molecular mechanisms of probiotics, the effect of probiotic mixtures versus single strains, the effect of formulation of probiotics and the fate of ingested probiotics should help to clarify the value of immune assays as selection criteria for probiotics.

scholarly journals Plectin ensures intestinal epithelial integrity and protects colon against colitis

2021 ◽  
Vol 14 (3) ◽  
pp. 691-702
Author(s):  
Alzbeta Krausova ◽  
Petra Buresova ◽  
Lenka Sarnova ◽  
Gizem Oyman-Eyrilmez ◽  
Jozef Skarda ◽  
...  
Keyword(s):  
Mechanical Stability ◽  
Intestinal Barrier ◽  
Intercellular Junctions ◽  
Protein Profiling ◽  
Cell Junctions ◽  
Intestinal Barrier Function ◽  
Intestinal Epithelial ◽  
Critical Determinant ◽  
In Vitro Feeding

AbstractPlectin, a highly versatile cytolinker protein, provides tissues with mechanical stability through the integration of intermediate filaments (IFs) with cell junctions. Here, we hypothesize that plectin-controlled cytoarchitecture is a critical determinant of the intestinal barrier function and homeostasis. Mice lacking plectin in an intestinal epithelial cell (IEC; PleΔIEC) spontaneously developed colitis characterized by extensive detachment of IECs from the basement membrane (BM), increased intestinal permeability, and inflammatory lesions. Moreover, plectin expression was reduced in the colons of ulcerative colitis (UC) patients and negatively correlated with the severity of colitis. Mechanistically, plectin deficiency in IECs led to aberrant keratin filament (KF) network organization and the formation of dysfunctional hemidesmosomes (HDs) and intercellular junctions. In addition, the hemidesmosomal α6β4 integrin (Itg) receptor showed attenuated association with KFs, and protein profiling revealed prominent downregulation of junctional constituents. Consistent with the effects of plectin loss in the intestinal epithelium, plectin-deficient IECs exhibited remarkably reduced mechanical stability and limited adhesion capacity in vitro. Feeding mice with a low-residue liquid diet that reduced mechanical stress and antibiotic treatment successfully mitigated epithelial damage in the PleΔIEC colon.

The Effects of Sulfated Secondary Bile Acids on Intestinal Barrier Function and Immune Response in an Inflammatory in vitro Human Intestinal Model

2021 ◽  
Author(s):  
Benthe van der Lugt ◽  
Maartje C.P. Vos ◽  
Mechteld Grootte Bromhaar ◽  
Noortje Ijssennagger ◽  
Frank Vrieling ◽  
...  
Keyword(s):  
Immune Response ◽  
Bile Acids ◽  
Barrier Function ◽  
Intestinal Barrier ◽  
Intestinal Barrier Function ◽  
Secondary Bile Acids

scholarly journals In vitro study of Lactobacillus plantarum properties as a potential probiotic strain and an alternative method to antibiotic treatment of fish

2018 ◽  
Vol 26 (1) ◽  
pp. 47-55 ◽  
Author(s):  
Barbara Kazuń ◽  
Krzysztof Kazuń ◽  
Joanna Żylińska ◽  
Andrzej K. Siwicki
Keyword(s):  
Lactobacillus Plantarum ◽  
Antibiotic Treatment ◽  
Intestinal Flora ◽  
In Vitro Study ◽  
Probiotic Strain ◽  
Aeromonas Salmonicida ◽  
Probiotic Properties ◽  
Opportunistic Bacteria ◽  
Fish Bile

Abstract The presence of lactic acid bacteria (LAB) favors the stabilization of intestinal flora, facilitates digestion, improves the assimilability of fodder, and has an immunomodulatory effect on the immune system. According to current research, the application of LAB following antibiotic treatment prevents the development of opportunistic bacteria inhabiting the digestive tract. In the study the potential probiotic properties of Lactobacillus plantarum strains, which can be administered as an alternative to antibiotic treatment in aquaculture, were investigated under in vitro conditions. The strains of L. plantarum were characterized for important properties such as the ability to grow in the presence of 10% fish bile, a tolerance of low pH, and antagonism to pathogens dangerous for fish such as Aeromonas salmonicida and Pseudomonas fluorescens; therefore, they meeting the criteria for strains with probiotic properties. In view of currently increasing resistance to antibiotics and a decrease of their efficiency, probiotic bacteria can serve to support immunity to infections in the future.

scholarly journals ISOLATION, CHARACTERIZATION AND IDENTIFICATION OF Lactobacillus fermentum EIPW5A ISOLATED FROM WHEY

2021 ◽  
pp. e248
Author(s):  
Dipanwita Bhattacharjee ◽  
Barun Bhattacharyya
Keyword(s):  
Lactic Acid Production ◽  
Probiotic Strain ◽  
16S Rrna Gene Sequencing ◽  
Lactobacillus Fermentum ◽  
Resistance Pattern ◽  
Rrna Gene ◽  
Probiotic Properties ◽  
Natural Habitats ◽  
Acid And Bile Tolerance ◽  
Rrna Gene Sequencing

The probiotic organisms are now used widely for different clinical indications. In an attempt to isolate a good probiotic strain for therapeutic applications, we have screened several isolates having probiotic attributes. The essential probiotic characters such as lactic acid production, antimicrobial activity, acid and bile tolerance, vitamin B12 production and antibiotic resistance pattern were considered as parameters for screening of probiotic bacteria from its natural habitats. Considering the said probiotic properties the strain EIPW5A was selected for the present study. The organism was identified as Lactobacillus fermentum based on its morphological, biochemical, physiological characters and 16S rRNA gene sequencing results.

scholarly journals Application of A Novel Potential Probiotic Lactobacillus paracasei Strain Isolated from Kefir Grains in the Production of Feta-Type Cheese

2018 ◽  
Vol 6 (4) ◽  
pp. 121 ◽  
Author(s):  
Ioanna Mantzourani ◽  
Antonia Terpou ◽  
Athanasios Alexopoulos ◽  
Pelagia Chondrou ◽  
Alex Galanis ◽  
...  
Keyword(s):  
Starter Culture ◽  
Quality Characteristics ◽  
Lactobacillus Paracasei ◽  
Pcr Analysis ◽  
In Vitro Tests ◽  
Probiotic Properties ◽  
The Novel ◽  
Kefir Grains ◽  
Cheese Production

In the present study 38 lactic acid bacteria strains were isolated from kefir grains and were monitored regarding probiotic properties in a series of established in vitro tests, including resistance to low pH, resistance to pepsin and pancreatin, and tolerance to bile salts, as well as susceptibility against common antibiotics. Among them, the strain SP3 displayed potential probiotic properties. Multiplex PCR analysis indicated that the novel strain belongs to the paracasei species. Likewise, the novel strain (Lactobacillus paracasei SP3) was applied as a starter culture for Feta-type cheese production. Feta-type cheese production resulted in significantly higher acidity; lower pH; reduced counts of coliforms, yeasts and fungi; and improved quality characteristics compared with cheese samples produced with no starter culture. Finally, it is highlighted that the application of the novel strain led to Feta-type cheese production with improved overall quality and sensory characteristics.

scholarly journals Removal of Antibiotic Resistance Gene-Carrying Plasmids from Lactobacillus reuteri ATCC 55730 and Characterization of the Resulting Daughter Strain, L. reuteri DSM 17938

2008 ◽  
Vol 74 (19) ◽  
pp. 6032-6040 ◽  
Author(s):  
Anna Rosander ◽  
Eamonn Connolly ◽  
Stefan Roos
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Lactobacillus Reuteri ◽  
Antibiotic Resistance Gene ◽  
Genetically Modify ◽  
Antibiotic Resistance Genes ◽  
Probiotic Strain ◽  
Probiotic Properties ◽  
Food Borne

ABSTRACT The spread of antibiotic resistance in pathogens is primarily a consequence of the indiscriminate use of antibiotics, but there is concern that food-borne lactic acid bacteria may act as reservoirs of antibiotic resistance genes when distributed in large doses to the gastrointestinal tract. Lactobacillus reuteri ATCC 55730 is a commercially available probiotic strain which has been found to harbor potentially transferable resistance genes. The aims of this study were to define the location and nature of β-lactam, tetracycline, and lincosamide resistance determinants and, if they were found to be acquired, attempt to remove them from the strain by methods that do not genetically modify the organism before subsequently testing whether the probiotic characteristics were retained. No known β-lactam resistance genes was found, but penicillin-binding proteins from ATCC 55730, two additional resistant strains, and three sensitive strains of L. reuteri were sequenced and comparatively analyzed. The β-lactam resistance in ATCC 55730 is probably caused by a number of alterations in the corresponding genes and can be regarded as not transferable. The strain was found to harbor two plasmids carrying tet(W) tetracycline and lnu(A) lincosamide resistance genes, respectively. A new daughter strain, L. reuteri DSM 17938, was derived from ATCC 55730 by removal of the two plasmids, and it was shown to have lost the resistances associated with them. Direct comparison of the parent and daughter strains for a series of in vitro properties and in a human clinical trial confirmed the retained probiotic properties of the daughter strain.

scholarly journals Lubiprostone Induces Claudin-1 and Protects Intestinal Barrier Function

Pharmacology ◽  
2019 ◽  
Vol 105 (1-2) ◽  
pp. 102-108 ◽  
Author(s):  
Norio Nishii ◽  
Tadayuki Oshima ◽  
Min Li ◽  
Hirotsugu Eda ◽  
Kumiko Nakamura ◽  
...  
Keyword(s):  
Barrier Function ◽  
Intestinal Barrier ◽  
Fluorescein Isothiocyanate ◽  
Intestinal Barrier Function ◽  
Epithelial Barrier ◽  
Dependent Manner ◽  
Epithelial Permeability ◽  
Epithelial Barrier Function ◽  
Dose Dependent

Introduction: Lubiprostone, a chloride channel activator, is said to reduce epithelial permeability. However, whether lubiprostone has a direct effect on the epithelial barrier function and how it modulates the intestinal barrier function remain unknown. Therefore, the effects of lubiprostone on intestinal barrier function were evaluated in vitro. Methods: Caco-2 cells were used to assess the intestinal barrier function. To examine the expression of claudins, immunoblotting was performed with specific antibodies. The effects of lubiprostone on cytokines (IFNγ, IL-6, and IL-1β) and aspirin-induced epithelial barrier disruption were assessed by transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC) labeled-dextran permeability. Results: IFNγ, IL-6, IL-1β, and aspirin significantly decreased TEER and increased epithelial permeability. Lubiprostone significantly improved the IFNγ-induced decrease in TEER in a dose-dependent manner. Lubiprostone significantly reduced the IFNγ-induced increase in FITC labeled-dextran permeability. The changes induced by IL-6, IL-1β, and aspirin were not affected by lubiprostone. The expression of claudin-1, but not claudin-3, claudin-4, occludin, and ZO-1 was significantly increased by lubiprostone. Conclusion: Lubiprostone significantly improved the IFNγ-induced decrease in TEER and increase in FITC labeled-dextran permeability. Lubiprostone increased the expression of claudin-1, and this increase may be related to the effect of lubiprostone on the epithelial barrier function.

scholarly journals Role for diet in normal gut barrier function: developing guidance within the framework of food-labeling regulations

2019 ◽  
Vol 317 (1) ◽  
pp. G17-G39 ◽  
Author(s):  
Michael Camilleri ◽  
Barbara J. Lyle ◽  
Karen L. Madsen ◽  
Justin Sonnenburg ◽  
Kristin Verbeke ◽  
...  
Keyword(s):  
Barrier Function ◽  
Intestinal Barrier ◽  
Normal Function ◽  
Model Systems ◽  
Intestinal Barrier Function ◽  
Dietary Interventions ◽  
Labeling Regulations ◽  
Gut Barrier Function ◽  
And Function

A reduction in intestinal barrier function is currently believed to play an important role in pathogenesis of many diseases, as it facilitates passage of injurious factors such as lipopolysaccharide, peptidoglycan, whole bacteria, and other toxins to traverse the barrier to damage the intestine or enter the portal circulation. Currently available evidence in animal models and in vitro systems has shown that certain dietary interventions can be used to reinforce the intestinal barrier to prevent the development of disease. The relevance of these studies to human health is unknown. Herein, we define the components of the intestinal barrier, review available modalities to assess its structure and function in humans, and review the available evidence in model systems or perturbations in humans that diet can be used to fortify intestinal barrier function. Acknowledging the technical challenges and the present gaps in knowledge, we provide a conceptual framework by which evidence could be developed to support the notion that diet can reinforce human intestinal barrier function to restore normal function and potentially reduce the risk for disease. Such evidence would provide information on the development of healthier diets and serve to provide a framework by which federal agencies such as the US Food and Drug Administration can evaluate evidence linking diet with normal human structure/function claims focused on reducing risk of disease in the general public.

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