Cross-Feeding among Probiotic Bacterial Strains on Prebiotic Inulin Involves the Extracellularexo-Inulinase ofLactobacillus paracaseiStrain W20

ABSTRACTProbiotic gut bacteria employ specific metabolic pathways to degrade dietary carbohydrates beyond the capabilities of their human host. Here, we report how individual commercial probiotic strains degrade prebiotic (inulin type) fructans. First, a structural analysis of commercial fructose oligosaccharide-inulin samples was performed. These β-(2-1)-fructans differ in termination by either glucose (GF) or fructose (FF) residues, with a broad variation in the degrees of polymerization (DPs). The growth of individual probiotic bacteria on short-chain inulin (sc-inulin) (Frutafit CLR), a β-(2-1)-fructan (DP 2 to DP 40), was studied.Lactobacillus salivariusW57 and other bacteria grew relatively poorly on sc-inulin, with only fractions of DP 3 and DP 5 utilized, reflecting uptake via specific transport systems followed by intracellular metabolism.Lactobacillus paracaseisubsp.paracaseiW20 completely used all sc-inulin components, employing an extracellularexo-inulinase enzyme (glycoside hydrolase family GH32 [LpGH32], also found in other strains of this species); the purified enzyme converted high-DP compounds into fructose, sucrose, 1-kestose, and F2 (inulobiose). The cocultivation ofL. salivariusW57 andL. paracaseiW20 on sc-inulin resulted in cross-feeding of the former by the latter, supported by this extracellularexo-inulinase. The extent of cross-feeding depended on the type of fructan, i.e., the GF type (clearly stimulating) versus the FF type (relatively low stimulus), and on fructan chain length, since relatively low-DP β-(2-1)-fructans contain a relatively high content of GF-type molecules, thus resulting in higher concentrations of GF-type DP 2 to DP 3 degradation products. The results provide an example of howin vivocross-feeding on prebiotic β-(2-1)-fructans may occur among probiotic lactobacilli.IMPORTANCEThe human gut microbial community is associated strongly with host physiology and human diseases. This observation has prompted research on pre- and probiotics, two concepts enabling specific changes in the composition of the human gut microbiome that result in beneficial effects for the host. Here, we show how fructooligosaccharide-inulin prebiotics are fermented by commercial probiotic bacterial strains involving specific sets of enzymes and transporters. Cross-feeding strains such asLactobacillus paracaseiW20 may thus act as keystone strains in the degradation of prebiotic inulin in the human gut, and this strain–exo-inulinase combination may be used in commercialLactobacillus-inulin synbiotics.

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Effects of Xylo-Oligosaccharides on Broiler Chicken Performance and Microbiota

Applied and Environmental Microbiology ◽

10.1128/aem.01616-15 ◽

2015 ◽

Vol 81 (17) ◽

pp. 5880-5888 ◽

Cited By ~ 74

Author(s):

C. De Maesschalck ◽

V. Eeckhaut ◽

L. Maertens ◽

L. De Lange ◽

L. Marchal ◽

...

Keyword(s):

Broiler Chickens ◽

Degradation Products ◽

Hydrolytic Degradation ◽

Feed Additives ◽

Gut Health ◽

Feed Conversion ◽

Content Type ◽

Beneficial Effects ◽

Coa Transferase

ABSTRACTIn broiler chickens, feed additives, including prebiotics, are widely used to improve gut health and to stimulate performance. Xylo-oligosaccharides (XOS) are hydrolytic degradation products of arabinoxylans that can be fermented by the gut microbiota. In the current study, we aimed to analyze the prebiotic properties of XOS when added to the broiler diet. Administration of XOS to chickens, in addition to a wheat-rye-based diet, significantly improved the feed conversion ratio. XOS significantly increased villus length in the ileum. It also significantly increased numbers of lactobacilli in the colon andClostridiumcluster XIVa in the ceca. Moreover, the number of gene copies encoding the key bacterial enzyme for butyrate production, butyryl-coenzyme A (butyryl-CoA):acetate CoA transferase, was significantly increased in the ceca of chickens administered XOS. In this group of chickens, at the species level,Lactobacillus crispatusandAnaerostipes butyraticuswere significantly increased in abundance in the colon and cecum, respectively.In vitrofermentation of XOS revealed cross-feeding betweenL. crispatusandA. butyraticus. Lactate, produced byL. crispatusduring XOS fermentation, was utilized by the butyrate-producingAnaerostipesspecies. These data show the beneficial effects of XOS on broiler performance when added to the feed, which potentially can be explained by stimulation of butyrate-producing bacteria through cross-feeding of lactate and subsequent effects of butyrate on gastrointestinal function.

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Differential Metabolism of Exopolysaccharides from Probiotic Lactobacilli by the Human Gut Symbiont Bacteroides thetaiotaomicron

Applied and Environmental Microbiology ◽

10.1128/aem.00149-15 ◽

2015 ◽

Vol 81 (12) ◽

pp. 3973-3983 ◽

Cited By ~ 30

Author(s):

Alicia Lammerts van Bueren ◽

Aakanksha Saraf ◽

Eric C. Martens ◽

Lubbert Dijkhuizen

Keyword(s):

Gastrointestinal Tract ◽

Carbohydrate Metabolism ◽

Lactobacillus Reuteri ◽

Bacterial Species ◽

Transport Systems ◽

Bacteroides Thetaiotaomicron ◽

Human Gut ◽

Positive Health ◽

Content Type ◽

Commensal Species

ABSTRACTProbiotic microorganisms are ingested as food or supplements and impart positive health benefits to consumers. Previous studies have indicated that probiotics transiently reside in the gastrointestinal tract and, in addition to modulating commensal species diversity, increase the expression of genes for carbohydrate metabolism in resident commensal bacterial species. In this study, it is demonstrated that the human gut commensal speciesBacteroides thetaiotaomicronefficiently metabolizes fructan exopolysaccharide (EPS) synthesized by probioticLactobacillus reuteristrain 121 while only partially degrading reuteran and isomalto/malto-polysaccharide (IMMP) α-glucan EPS polymers.B. thetaiotaomicronmetabolized these EPS molecules via the activation of enzymes and transport systems encoded by dedicated polysaccharide utilization loci specific for β-fructans and α-glucans. Reduced metabolism of reuteran and IMMP α-glucan EPS molecules may be due to reduced substrate binding by components of the starch utilization system (sus). This study reveals that microbial EPS substrates activate genes for carbohydrate metabolism inB. thetaiotaomicronand suggests that microbially derived carbohydrates provide a carbohydrate-rich reservoir forB. thetaiotaomicronnutrient acquisition in the gastrointestinal tract.

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Qualifying the T-2 Toxin-Degrading Properties of Seven Microbes with Zebrafish Embryo Microinjection Method

Toxins ◽

10.3390/toxins12070460 ◽

2020 ◽

Vol 12 (7) ◽

pp. 460

Author(s):

Edina Garai ◽

Anita Risa ◽

Emese Varga ◽

Mátyás Cserháti ◽

Balázs Kriszt ◽

...

Keyword(s):

Sublethal Effects ◽

Degradation Products ◽

Rhodococcus Erythropolis ◽

Bacterial Strains ◽

Cell Stage ◽

Bacterial Metabolites ◽

Mycotoxin Degradation

T-2 mycotoxin degradation and detoxification efficiency of seven bacterial strains were investigated with zebrafish microinjection method in three steps ((1) determination of mycotoxin toxicity baseline, (2) examination of bacterial metabolites toxicity, (3) identification of degradation products toxicity). Toxicity of T-2 was used as a baseline of toxic effects, bacterial metabolites of strains as control of bacterial toxicity and degradation products of toxin as control of biodegradation were injected into one-cell stage embryos in the same experiment. The results of in vivo tests were checked and supplemented with UHPLC-MS/MS measurement of T-2 concentration of samples. Results showed that the Rhodococcus erythropolis NI1 strain was the only one of the seven tested (R. gordoniae AK38, R. ruber N361, R. coprophilus N774, R. rhodochrous NI2, R. globerulus N58, Gordonia paraffinivorans NZS14), which was appropriated to criteria all aspects (bacterial and degradation metabolites of strains caused lower toxicity effects than T-2, and strains were able to degrade T-2 mycotoxin). Bacterial and degradation metabolites of the NI1 strain caused slight lethal and sublethal effects on zebrafish embryos at 72- and 120-h postinjection. Results demonstrated that the three-step zebrafish microinjection method is well-suited to the determination and classification of different bacterial strains by their mycotoxin degradation and detoxification efficiency.

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Survival and beneficial effects of propionibacteria in the human gut: in vivo and in vitro investigations

Dairy Science & Technology ◽

10.1051/lait:2001012 ◽

2002 ◽

Vol 82 (1) ◽

pp. 131-144 ◽

Cited By ~ 41

Author(s):

Gw�na�l Jan ◽

Pauline Leverrier ◽

Isabelle Proudy ◽

Nathalie Roland

Keyword(s):

Human Gut ◽

Beneficial Effects

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Even Apparently Insignificant Chemical Deviations among Bioequivalent Generic Antibiotics Can Lead to Therapeutic Nonequivalence: the Case of Meropenem

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00350-13 ◽

2013 ◽

Vol 58 (2) ◽

pp. 1005-1018 ◽

Cited By ~ 16

Author(s):

M. Agudelo ◽

C. A. Rodriguez ◽

C. A. Pelaez ◽

O. Vesga

Keyword(s):

Guinea Pig ◽

Susceptibility Testing ◽

Degradation Products ◽

Active Pharmaceutical Ingredient ◽

Infection Model ◽

Content Type ◽

Infection Models ◽

Generic Products

ABSTRACTSeveral studies with animal models have demonstrated that bioequivalence of generic products of antibiotics like vancomycin, as currently defined, do not guarantee therapeutic equivalence. However, the amounts and characteristics of impurities and degradation products in these formulations do not violate the requirements of the U.S. Pharmacopeia (USP). Here, we provide experimental data with three generic products of meropenem that help in understanding how these apparently insignificant chemical differences affect thein vivoefficacy. Meropenem generics were compared with the innovatorin vitroby microbiological assay, susceptibility testing, and liquid chromatography/mass spectrometry (LC/MS) analysis andin vivowith the neutropenic guinea pig soleus infection model (Pseudomonas aeruginosa) and the neutropenic mouse thigh (P. aeruginosa), brain (P. aeruginosa), and lung (Klebisella pneumoniae) infection models, adding the dihydropeptidase I (DHP-I) inhibitor cilastatin in different proportions to the carbapenem. We found that the concentration and potency of the active pharmaceutical ingredient,in vitrosusceptibility testing, and mouse pharmacokinetics were identical for all products; however, two generics differed significantly from the innovator in the guinea pig and mouse models, while the third generic was therapeutically equivalent under all conditions. Trisodium adducts in a bioequivalent generic made it more susceptible to DHP-I hydrolysis and less stable at room temperature, explaining its therapeutic nonequivalence. We conclude that the therapeutic nonequivalence of generic products of meropenem is due to greater susceptibility to DHP-I hydrolysis. These failing generics are compliant with USP requirements and would remain undetectable under current regulations.

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Draft Genome Sequence of the Feruloyl Esterase-Producing Strain Lactobacillus fermentum CRL1446, a Probiotic for Malnutrition

Genome Announcements ◽

10.1128/genomea.00225-18 ◽

2018 ◽

Vol 6 (21) ◽

Cited By ~ 1

Author(s):

María C. Abeijón Mukdsi ◽

Lucila Saavedra ◽

María P. Gauffin Cano ◽

Elvira M. Hebert ◽

Roxana B. Medina

Keyword(s):

Genome Sequence ◽

Draft Genome ◽

Lactobacillus Fermentum ◽

Feruloyl Esterase ◽

Draft Genome Sequence ◽

Probiotic Properties ◽

Content Type ◽

Beneficial Effects ◽

Interesting Candidate

ABSTRACT We report here the draft genome sequence of Lactobacillus fermentum CRL1446 (2,148,781 bp, 51.4% G+C content). This strain exhibits feruloyl esterase activity and important technological and probiotic properties. Because of its proven beneficial effects in vivo, it represents an interesting candidate for the development of functional foods or pharmabiotics for malnutrition.

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Differential Targeting of the E-Cadherin/β-Catenin Complex by Gram-Positive Probiotic Lactobacilli Improves Epithelial Barrier Function

Applied and Environmental Microbiology ◽

10.1128/aem.06983-11 ◽

2011 ◽

Vol 78 (4) ◽

pp. 1140-1147 ◽

Cited By ~ 48

Author(s):

Stephanie Hummel ◽

Katharina Veltman ◽

Christoph Cichon ◽

Ulrich Sonnenborn ◽

M. Alexander Schmidt

Keyword(s):

Barrier Function ◽

Epithelial Barrier ◽

Gram Positive ◽

Content Type ◽

Epithelial Barrier Function ◽

Beneficial Effects ◽

Gastrointestinal Barrier ◽

Probiotic Lactobacilli ◽

Adherence Junction ◽

Junction Proteins

ABSTRACTThe intestinal ecosystem is balanced by dynamic interactions between resident and incoming microbes, the gastrointestinal barrier, and the mucosal immune system. However, in the context of inflammatory bowel diseases (IBD), where the integrity of the gastrointestinal barrier is compromised, resident microbes contribute to the development and perpetuation of inflammation and disease. Probiotic bacteria have been shown to exert beneficial effects, e.g., enhancing epithelial barrier integrity. However, the mechanisms underlying these beneficial effects are only poorly understood. Here, we comparatively investigated the effects of four probiotic lactobacilli, namely,Lactobacillus acidophilus,L. fermentum,L. gasseri, andL. rhamnosus, in a T84 cell epithelial barrier model. Results of DNA microarray experiments indicating that lactobacilli modulate the regulation of genes encoding in particular adherence junction proteins such as E-cadherin and β-catenin were confirmed by quantitative reverse transcription-PCR (qRT-PCR). Furthermore, we show that epithelial barrier function is modulated by Gram-positive probiotic lactobacilli via their effect on adherence junction protein expression and complex formation. In addition, incubation with lactobacilli differentially influences the phosphorylation of adherence junction proteins and the abundance of protein kinase C (PKC) isoforms such as PKCδ that thereby positively modulates epithelial barrier function. Further insight into the underlying molecular mechanisms triggered by these probiotics might also foster the development of novel strategies for the treatment of gastrointestinal diseases (e.g., IBD).

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Synthetic Gene Circuits Enable Systems-Level Biosensor Trigger Discovery at the Host-Microbe Interface

mSystems ◽

10.1128/msystems.00125-19 ◽

2019 ◽

Vol 4 (4) ◽

Cited By ~ 11

Author(s):

Alexander D. Naydich ◽

Shannon N. Nangle ◽

Johannes J. Bues ◽

Disha Trivedi ◽

Nabeel Nissar ◽

...

Keyword(s):

Intestinal Inflammation ◽

Intestinal Bacteria ◽

Bacterial Strains ◽

Bacterial Biosensor ◽

Content Type ◽

Memory Circuit ◽

Treatment And Prevention ◽

Engineered Systems ◽

Sense And Respond

ABSTRACTEngineering synthetic circuits into intestinal bacteria to sense, record, and respond toin vivosignals is a promising new approach for the diagnosis, treatment, and prevention of disease. However, because the design of disease-responsive circuits is limited by a relatively small pool of known biosensors, there is a need for expanding the capacity of engineered bacteria to sense and respond to the host environment. Here, we apply a robust genetic memory circuit inEscherichia colito identify new bacterial biosensor triggers responding in the healthy and diseased mammalian gut, which may be used to construct diagnostic or therapeutic circuits. We developed a pipeline for rapid systems-level library construction and screening, using next-generation sequencing and computational analysis, which demonstrates remarkably reliable identification of responsive biosensor triggers from pooled libraries. By testing libraries of potential triggers—each consisting of a promoter and ribosome binding site (RBS)—and using RBS variation to augment the range of trigger sensitivity, we identify and validate triggers that selectively activate our synthetic memory circuit during transit through the gut. We further identify biosensor triggers with increased response in the inflamed gut through comparative screening of one of our libraries in healthy mice and those with intestinal inflammation. Our results demonstrate the power of systems-level screening for the identification of novel biosensor triggers in the gut and provide a platform for disease-specific screening that is capable of contributing to both the understanding and clinical management of intestinal illness.IMPORTANCEThe gut is a largely obscure and inaccessible environment. The use of live, engineered probiotics to detect and respond to disease signalsin vivorepresents a new frontier in the management of gut diseases. Engineered probiotics have also shown promise as a novel mechanism for drug delivery. However, the design and construction of effective strains that respond to thein vivoenvironment is hindered by our limited understanding of bacterial behavior in the gut. Our work expands the pool of environmentally responsive synthetic circuits for the healthy and diseased gut, providing insight into host-microbe interactions and enabling future development of increasingly complex biosensors. This method also provides a framework for rapid prototyping of engineered systems and for application across bacterial strains and disease models, representing a practical step toward the construction of clinically useful synthetic tools.

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Dietary Nutrients, Proteomes, and Adhesion of Probiotic Lactobacilli to Mucin and Host Epithelial Cells

Microorganisms ◽

10.3390/microorganisms6030090 ◽

2018 ◽

Vol 6 (3) ◽

pp. 90 ◽

Cited By ~ 7

Author(s):

Hasan Celebioglu ◽

Birte Svensson

Keyword(s):

Epithelial Cells ◽

Probiotic Bacteria ◽

In Vitro Techniques ◽

Beneficial Effects ◽

Food Components ◽

Dietary Nutrients ◽

Probiotic Lactobacilli ◽

Host Epithelium

The key role of diet and environment in human health receives increasing attention. Thus functional foods, probiotics, prebiotics, and synbiotics with beneficial effects on health and ability to prevent diseases are in focus. The efficacy of probiotic bacteria has been connected with their adherence to the host epithelium and residence in the gut. Several in vitro techniques are available for analyzing bacterial interactions with mucin and intestinal cells, simulating adhesion to the host in vivo. Proteomics has monitored and identified proteins of probiotic bacteria showing differential abundance elicited in vitro by exposure to food components, including potential prebiotics (e.g., certain carbohydrates, and plant polyphenols). While adhesion of probiotic bacteria influenced by various environmental factors relevant to the gastrointestinal tract has been measured previously, this was rarely correlated with changes in the bacterial proteome induced by dietary nutrients. The present mini-review deals with effects of selected emerging prebiotics, food components and ingredients on the adhesion of probiotic lactobacilli to mucin and gut epithelial cells and concomitant abundancy changes of specific bacterial proteins. Applying this in vitro synbiotics-like approach enabled identification of moonlighting and other surface-located proteins of Lactobacillus acidophilus NCFM that are possibly associated with the adhesive mechanism.

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Characterization and Functional Test of Canine Probiotics

Frontiers in Microbiology ◽

10.3389/fmicb.2021.625562 ◽

2021 ◽

Vol 12 ◽

Author(s):

Hyun-Jun Jang ◽

Seungwoo Son ◽

Jung-Ae Kim ◽

Min Young Jung ◽

Yeon-jae Choi ◽

...

Keyword(s):

Animal Health ◽

Clinical Results ◽

Industrial Applications ◽

Bacterial Strains ◽

Host Animal ◽

Microbial Composition ◽

Beneficial Effects ◽

Acid And Bile Tolerance

Probiotics can modulate the composition of gut microbiota and benefit the host animal health in multiple ways. Lactic acid bacteria (LAB), mainly Lactobacillus and Bifidobacterium species, are well-known microbes with probiotic potential. In the present study, 88 microbial strains were isolated from canine feces and annotated. Among these, the four strains CACC517, 537, 558, and 566 were tested for probiotic characteristics, and their beneficial effects on hosts were evaluated both in vitro and in vivo; these strains exhibited antibiosis, antibiotic activity, acid and bile tolerance, and relative cell adhesion to the HT-29 monolayer cell line. Byproducts of these strains increased the viability and decreased oxidative stress in mouse and dog cell lines (RAW264.7 and DH82, respectively). Subsequently, when the probiotics were applied to the clinical trial, changes in microbial composition and relative abundance of bacterial strains were clearly observed in the experimental animals. Experimental groups before and after the application were obviously separated from PCA analysis of clinical results. Conclusively, these results could provide comprehensive understanding of the effects of probiotic strains (CACC517, 537, 558, and 566) and their industrial applications.

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