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Gut Dec 2021Using faecal shotgun metagenomic sequencing, we identified the depletion of in patients with colorectal cancer (CRC). We aimed to determine the potential...
OBJECTIVE
Using faecal shotgun metagenomic sequencing, we identified the depletion of in patients with colorectal cancer (CRC). We aimed to determine the potential antitumourigenic role of in colorectal tumourigenesis.
DESIGN
The tumor-suppressive effect of was assessed in murine models of CRC. CRC cell lines and organoids derived from patients with CRC were cultured with or MG1655 culture-supernatant to evaluate cell proliferation, apoptosis and cell cycle distribution. Gut microbiota was assessed by 16S ribosomal DNA sequencing. Antitumour molecule produced from was identified by liquid chromatography mass spectrometry (LC-MS/MS) and targeted mass spectrometry.
RESULTS
significantly reduced intestinal tumour number and size compared with MG1655 and phosphate-buffered saline in both male and female murine intestinal tumourigenesis models. Faecal microbial profiling revealed enrichment of probiotics and depletion of pathogenic bacteria in -treated mice. Culturing CRC cells with culture-supernatant (5%, 10% and 20%) concentration-dependently suppressed cell proliferation and colony formation. culture-supernatant significantly promoted apoptosis in CRC cells and patient-derived CRC organoids, but not in normal colon epithelial cells. Only culture-supernatant with fraction size <3 kDa suppressed proliferation in CRC cells. Using LC-MS/MS, enrichments of indole-3-lactic acid (ILA) was identified in both culture-supernatant and the gut of -treated mice. ILA displayed anti-CRC growth and inhibited intestinal tumourigenesis .
CONCLUSION
protects against intestinal tumourigenesis by producing protective metabolites that can promote apoptosis of CRC cells.
PubMed: 34937766
DOI: 10.1136/gutjnl-2020-323951 -
Gut Nov 2023Gut microbiota is a key player in dictating immunotherapy response. We aimed to explore the immunomodulatory effect of probiotic and its role in improving...
OBJECTIVE
Gut microbiota is a key player in dictating immunotherapy response. We aimed to explore the immunomodulatory effect of probiotic and its role in improving anti-programmed cell death protein 1 (PD1) efficacy against colorectal cancer (CRC).
DESIGN
The effects of in anti-PD1 response were assessed in syngeneic mouse models and azoxymethane/dextran sulfate sodium-induced CRC model. The change of immune landscape was identified by multicolour flow cytometry and validated by immunohistochemistry staining and in vitro functional assays. Liquid chromatography-mass spectrometry was performed to identify the functional metabolites.
RESULTS
significantly improved anti-PD1 efficacy in two syngeneic mouse models with different microsatellite instability (MSI) statuses (MSI-high for MC38, MSI-low for CT26). Such effect was confirmed in CRC tumourigenesis model. synergised with anti-PD1 therapy by reducing Foxp3 CD25 regulatory T cell (Treg) intratumoural infiltration, and enhancing effector function of CD8 T cells. -derived indole-3-carboxylic acid (ICA) was identified as the functional metabolite. Mechanistically, ICA inhibited indoleamine 2,3-dioxygenase (IDO1) expression, therefore suppressing kynurenine (Kyn) production in tumours. ICA also competed with Kyn for binding site on aryl hydrocarbon receptor (AHR) and antagonised Kyn binding on CD4 T cells, thereby inhibiting Treg differentiation in vitro. ICA phenocopied effect and significantly improved anti-PD1 efficacy in vivo, which could be reversed by Kyn supplementation.
CONCLUSION
-derived ICA improved anti-PD1 efficacy in CRC through suppressing CD4+Treg differentiation and enhancing CD8+T cell function by modulating the IDO1/Kyn/AHR axis. is a potential adjuvant to augment anti-PD1 efficacy against CRC.
Topics: Animals; Mice; CD8-Positive T-Lymphocytes; Colorectal Neoplasms; Kynurenine; Receptors, Aryl Hydrocarbon; T-Lymphocytes, Regulatory; Lactobacillus; Programmed Cell Death 1 Receptor; Immune Checkpoint Inhibitors; Bacterial Lysates
PubMed: 37770127
DOI: 10.1136/gutjnl-2023-329543 -
Poultry Science Jan 2019Although Lactobacillus species have been administered widely as probiotics in poultry production, the mechanisms responsible for their functionality are not well...
Although Lactobacillus species have been administered widely as probiotics in poultry production, the mechanisms responsible for their functionality are not well understood. The genetic tools available for use in lactobacilli are advanced but have not been applied widely to investigate their probiotic functionality in poultry. The genome sequence of Lactobacillus gallinarum ATCC 33199, originally isolated from the chicken crop, has recently been made available suggesting this organism as a potentially important model organism for probiotic research in poultry. In this study, we demonstrated the functionality of the pORI28 system for construction of isogenic knockout mutants in L. gallinarum ATCC 33199 using insertional inactivation of lacL as proof-of-principle. The establishment of an effective chromosomal integration system for use in L. gallinarum ATCC 33199 will provide a platform for functional genomic analyses to investigate the functionality of this model organism in the gastrointestinal tract of poultry.
Topics: Chromosomes, Bacterial; Gene Knockout Techniques; Gene Silencing; Lactobacillus; Lactose; Mutagenesis, Insertional; beta-Galactosidase
PubMed: 30124967
DOI: 10.3382/ps/pey363 -
Current Microbiology Aug 1995The technique random amplified polymorphic DNA (RAPD)-PCR was evaluated as a method to differentiate Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus...
Evaluation of random amplified polymorphic DNA (RAPD)-PCR as a method to differentiate Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus amylovorus, Lactobacillus gallinarum, Lactobacillus gasseri, and Lactobacillus johnsonii.
The technique random amplified polymorphic DNA (RAPD)-PCR was evaluated as a method to differentiate Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus amylovorus, Lactobacillus gallinarum, Lactobacillus gasseri, and Lactobacillus johnsonii. Representative strains, including the type of each species, were selected from different clusters obtained by numerical analysis of total soluble cell protein patterns. Results obtained by RAPD-PCR corresponded well with results obtained by numerical analysis of total soluble cell protein patterns. The type strains of each species displayed different RAPD profiles. Strains with identical L(+)- nicotinamide adenine dinucleotide-dependent lactic dehydrogenase (nLDH) electrophoretic profiles could be distinguished on the basis of their RAPD profiles.
Topics: Base Sequence; DNA, Bacterial; Lactobacillus; Molecular Sequence Data; Polymerase Chain Reaction; Polymorphism, Genetic
PubMed: 7606186
DOI: 10.1007/BF00294286 -
Probiotics and Antimicrobial Proteins Feb 2021Pathogens develop resistance to antibiotics at a rate much faster than the discovery of new antimicrobial compounds. Reports of multidrug-resistant bacteria isolated... (Review)
Review
Pathogens develop resistance to antibiotics at a rate much faster than the discovery of new antimicrobial compounds. Reports of multidrug-resistant bacteria isolated from broilers, and the possibility that these strains may spread diseases amongst humans, prompted many European countries to ban the inclusion of antibiotics in feed. Probiotics added to broiler feed controlled a number of bacterial infections. A combination of Enterococcus faecium, Pediococcus acidilactici, Bacillus animalis, Lactobacillus salivarius and Lactobacillus reuteri decreased the colonisation of Campylobacter jejuni and Salmonella Enteritidis in the gastro-intestinal tract (GIT) of broilers, whereas Bacillus subtilis improved feed conversion, intestinal morphology, stimulated the immune system and inhibited the colonisation of Campylobacter jejuni, Escherichia coli and Salmonella Minnesota. Lactobacillus salivarius and Pediococcus parvulus improved weight gain, bone characteristics, intestinal morphology and immune response, and decreased the colonisation of S. Enteritidis. Lactobacillus crispatus, L. salivarius, Lactobacillus gallinarum, Lactobacillus johnsonii, Enterococcus faecalis and Bacillus amyloliquefaciens decreased the Salmonella count and led to an increase in lysozyme and T lymphocytes. Probiotics may also improve feed digestion through production of phytases, lipases, amylases and proteases or stimulate the GIT to secrete digestive enzymes. Some strains increase the nutritional value of feed by production of vitamins, exopolysaccharides and antioxidants. Bacteriocins, if produced, regulate pathogen numbers in the GIT and keep pro-inflammatory and anti-inflammatory reactions in balance.
Topics: Animal Feed; Animals; Anti-Bacterial Agents; Bacteria; Chickens; Gastrointestinal Microbiome; Probiotics
PubMed: 32556932
DOI: 10.1007/s12602-020-09640-z -
International Journal of Systematic... Jul 1992Biochemical properties and DNA-DNA reassociation studies of Lactobacillus acidophilus strains isolated from humans and animals indicate that these include six...
Taxonomic study of the Lactobacillus acidophilus group, with recognition of Lactobacillus gallinarum sp. nov. and Lactobacillus johnsonii sp. nov. and synonymy of Lactobacillus acidophilus group A3 (Johnson et al. 1980) with the type strain of Lactobacillus amylovorus (Nakamura 1981).
Biochemical properties and DNA-DNA reassociation studies of Lactobacillus acidophilus strains isolated from humans and animals indicate that these include six genomospecies. Two new species can be differentiated from the established species of the genus Lactobacillus: L. gallinarum sp. nov. (type strain, ATCC 33199) and L. johnsonii sp. nov. (type strain, ATCC 33200). Furthermore, it was clarified that L. acidophilus group A3 (Johnson et al. 1980) is synonymous with L. amylovorus.
Topics: DNA, Bacterial; Lactobacillus; Lactobacillus acidophilus; Nucleic Acid Hybridization; Phenotype; Sequence Homology, Nucleic Acid
PubMed: 1503977
DOI: 10.1099/00207713-42-3-487 -
Applied and Environmental Microbiology Feb 2014Probiotics have been demonstrated to promote growth, stimulate immune responses, and improve food safety of poultry. While widely used, their effectiveness is mixed, and...
Probiotics have been demonstrated to promote growth, stimulate immune responses, and improve food safety of poultry. While widely used, their effectiveness is mixed, and the mechanisms through which they contribute to poultry production are not well understood. Microbial phytases are increasingly supplemented in feed to improve digestibility and reduce antinutritive effects of phytate. The microbial origin of these exogenous enzymes suggests a potentially important mechanism of probiotic functionality. We investigated phytate degradation as a novel probiotic mechanism using recombinant Lactobacillus cultures expressing Bacillus subtilis phytase. B. subtilis phyA was codon optimized for expression in Lactobacillus and cloned into the expression vector pTRK882. The resulting plasmid, pTD003, was transformed into Lactobacillus acidophilus, Lactobacillus gallinarum, and Lactobacillus gasseri. SDS-PAGE revealed a protein in the culture supernatants of Lactobacillus pTD003 transformants with a molecular weight similar to that of the B. subtilis phytase. Expression of B. subtilis phytase increased phytate degradation of L. acidophilus, L. gasseri, and L. gallinarum approximately 4-, 10-, and 18-fold over the background activity of empty-vector transformants, respectively. Phytase-expressing L. gallinarum and L. gasseri were administered to broiler chicks fed a phosphorus-deficient diet. Phytase-expressing L. gasseri improved weight gain of broiler chickens to a level comparable to that for chickens fed a control diet adequate in phosphorus, demonstrating proof of principle that administration of phytate-degrading probiotic cultures can improve performance of livestock animals. This will inform future studies investigating whether probiotic cultures are able to provide both the performance benefits of feed enzymes and the animal health and food safety benefits traditionally associated with probiotics.
Topics: 6-Phytase; Animals; Bacillus subtilis; Body Weight; Chickens; Cloning, Molecular; Gene Expression; Lactobacillus; Phytic Acid; Probiotics; Recombinant Proteins
PubMed: 24271165
DOI: 10.1128/AEM.03155-13 -
Poultry Science Nov 2014Administration of probiotic Lactobacillus cultures is an important alternative to the use of antibiotic growth promoters and has been demonstrated to improve animal...
Administration of probiotic Lactobacillus cultures is an important alternative to the use of antibiotic growth promoters and has been demonstrated to improve animal health, growth performance, and preharvest food safety in poultry production. Whereas gastrointestinal colonization is thought to be critical to their probiotic functionality, factors important to Lactobacillus colonization in chickens are not well understood. In this study we investigate epithelial cell adhesion in vitro and colonization of Lactobacillusin vivo in broiler chickens. Adhesion of Lactobacillus cultures to epithelial cells was evaluated using the chicken LMH cell line. Lactobacillus cultures were able adhere effectively to LMH cells relative to Bacillus subtilis and Salmonella Typhimurium. Epithelial cell adhesion was similar for Lactobacillus crispatus TDCC 75, L. cristpatus TDCC 76, and Lactobacillus gallinarum TDCC 77, and all 3 were more adherent than L. gallinarum TDCC 78. However, when colonization was evaluated in the ileum and cecum of broiler chicks, L. crispatus TDCC 75 and L. gallinarum TDCC 77 were more persistent than L. crispatus TDCC 76 and L. gallinarum TDCC 78. The reduction of growth in medium supplemented with oxgal was greater for L. gallinarum TDCC 78 than L. gallinarum TDCC 77, suggesting that whereas adhesion was similar for the 2 strains, the difference in colonization between L. gallinarum strains may be due in part to their bile sensitivity. This study demonstrates that whereas adhesion to epithelial cells may be important in predicting gastrointestinal colonization, other factors including bile tolerance may also contribute to the colonization of Lactobacillus in poultry. Additionally, the chicken LMH cell line is expected to provide a platform for investigating mechanisms of Lactobacillus adhesion to epithelial tissue and evaluating the probiotic potential Lactobacillus in poultry.
Topics: Animal Feed; Animals; Bacterial Adhesion; Bile; Cattle; Cecum; Cell Line; Chickens; Diet; Epithelial Cells; Gastrointestinal Tract; Ileum; Lactobacillus; Male; Probiotics; Random Allocation
PubMed: 25239531
DOI: 10.3382/ps.2014-04076 -
Current Microbiology Apr 2014We propose a new method that allows accurate discrimination of Lactobacillus helveticus from other closely related homofermentative lactobacilli, especially...
Peptidoglycan hydrolases as species-specific markers to differentiate Lactobacillus helveticus from Lactobacillus gallinarum and other closely related homofermentative lactobacilli.
We propose a new method that allows accurate discrimination of Lactobacillus helveticus from other closely related homofermentative lactobacilli, especially Lactobacillus gallinarum. This method is based on the amplification by PCR of two peptidoglycan hydrolytic genes, Lhv_0190 and Lhv_0191. These genes are ubiquitous and show high homology at the intra-species level. The PCR method gave two specific PCR products, of 542 and 747 bp, for 25 L. helveticus strains coming from various sources. For L. gallinarum, two amplicons were obtained, the specific 542 bp amplicon and another one with a size greater than 1,500 bp. No specific PCR products were obtained for 12 other closely related species of lactobacilli, including the L. acidophilus complex, L. delbrueckii, and L. ultunensis. The developed PCR method provided rapid, precise, and easy identification of L. helveticus. Moreover, it enabled differentiation between the two closely phylogenetically related species L. helveticus and L. gallinarum.
Topics: DNA, Bacterial; Genetic Markers; Lactobacillus; N-Acetylmuramoyl-L-alanine Amidase; Phylogeny
PubMed: 24362553
DOI: 10.1007/s00284-013-0512-5 -
International Journal of Systematic and... Nov 2003Four strains isolated from chicken small intestine and strains JCM 1038 and JCM 1039 (designated as Lactobacillus acidophilus) were characterized by phenotypic and...
Four strains isolated from chicken small intestine and strains JCM 1038 and JCM 1039 (designated as Lactobacillus acidophilus) were characterized by phenotypic and molecular taxonomic methods. They were Gram-positive, catalase-negative, facultatively anaerobic rods that did not produce gas from glucose. These strains had similar phenotypic characteristics and exhibited intergroup DNA relatedness values of >77 %, indicating that they comprised a single species. The 16S rRNA gene sequence of a representative strain, JCM 1039(T) (designated as type strain in this study), was determined and aligned with those of other Lactobacillus species. JCM 1039(T) was placed in the Lactobacillus delbrueckii cluster of the genus Lactobacillus on the basis of phylogenetic analysis and formed an independent cluster that was distinct from its closest neighbours, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus gallinarum, L. acidophilus and Lactobacillus helveticus. Results of DNA-DNA hybridization experiments and whole-cell protein profiles clearly indicated that these strains represent a novel Lactobacillus species, for which the name Lactobacillus kitasatonis sp. nov. is proposed; the type strain of this species is JCM 1039(T).
Topics: Animals; Chickens; DNA, Bacterial; Genome, Bacterial; Glucose; Intestines; Lactobacillus; Lactobacillus acidophilus; Molecular Sequence Data; Phylogeny; Species Specificity
PubMed: 14657145
DOI: 10.1099/ijs.0.02815-0