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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 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 -
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 -
Viruses Nov 2022The cervical microbiota is essential in female sexual health, and its altered states seem to have a central role in the dynamic of high-risk papillomavirus (hrHPV)...
The cervical microbiota is essential in female sexual health, and its altered states seem to have a central role in the dynamic of high-risk papillomavirus (hrHPV) infections. This study aimed to evaluate the variation in bacterial communities' compositions according to hrHPV. We collected two samples per woman, with a difference of 12 ± 1 months between them, and performed a follow-up on 66 of these women. The viral load (VL) of the hrHPV was estimated by quantitative PCR (qPCR), then it was normalized (using the gene as reference) and transformed to the Log scale to facilitate the interpretation. The VL was categorized as Negative, without hrHPV copies; Low, less than 10 hrHPV copies; Medium, between 10 to 10 hrHPV copies; and High, >10 hrHPV copies. The microbiota composition was described through the Illumina Novaseq PE250 platform. The diversity analyses revealed changes regarding the hrHPV VL, where women with low VL (<10 hrHPV copies) presented high diversity. The community state type (CST) IV was the most common. However, in women with high VL, a lower association with depletion was found. and were the most abundant species in women with high VL, whereas women with low VL had a 6.06 greater probability of exhibiting dominance. We identified conspicuous differences in the abundance of 78 bacterial genera between women with low and high VL, where 26 were depleted (e.g., ) and 52 increased (e.g., ). A multilevel mixed-effects linear regression showed changes in the diversity due to the interaction between the measurement time and the VL, with a decrease in diversity in the second follow-up in women with low VL (Coeff. = 0.47), whereas the women with medium VL displayed an increase in diversity (Coeff. = 0.58). Here, we report for the first time that the cervical microbiota is influenced by the number of copies of hrHPV, where a decrease in the abundance of , greater diversity, and enrichment of bacterial taxa is relevant in women with low VL.
Topics: Female; Humans; Human Papillomavirus Viruses; Vagina; Papillomavirus Infections; Cervix Uteri; Microbiota; Papillomaviridae; Bacteria; Uterine Cervical Neoplasms
PubMed: 36560678
DOI: 10.3390/v14122674 -
Animals : An Open Access Journal From... Apr 2019Aflatoxin contamination in human food and animal feed is a threat to public safety. Aflatoxin B1 (AFB1) can be especially damaging to poultry production and consequently...
Aflatoxin contamination in human food and animal feed is a threat to public safety. Aflatoxin B1 (AFB1) can be especially damaging to poultry production and consequently economic development of Pakistan. The present study assessed the in vitro binding of AFB1 by indigenously characterized probiotic lactobacilli. Six isolates ( PDP 10, FYP 38, PDP 24, PL 53, PL 120, and PL 149) were tested for activity against toxigenic W-7.1 (AFB1 producer) by well diffusion assay. Only three isolates (PL 53, PL 120, and PL 149) had activity against W-7.1. The ameliorative effect of these probiotic isolates on AFB1 production was determined by co-culturing fungus with lactobacilli for 12 days, followed by aflatoxin quantification by high-performance liquid chromatography. In vitro AFB1 binding capacities of lactobacilli were determined by their incubation with a standard amount of AFB1 in phosphate buffer saline at 37 °C for 2 h. AFB1 binding capacities of isolates ranged from 28-65%. Four isolates (PDP 10, PDP 24, PL 120, and PL 149) also ceased aflatoxin production completely, whereas PL 53 showed 55% reduction in AFB1 production as compared to control. The present study demonstrated PL 149 to be an effective candidate AFB1 binding agent against . These findings further support the binding ability of lactic acid bacteria for dietary contaminants.
PubMed: 30991667
DOI: 10.3390/ani9040166 -
Journal of Dairy Science Mar 2015Lactobacillus gasseri is a widespread commensal lactic acid bacterium inhabiting human mucosal niches and has many beneficial effects as a probiotic. However, L. gasseri...
Lactobacillus gasseri is a widespread commensal lactic acid bacterium inhabiting human mucosal niches and has many beneficial effects as a probiotic. However, L. gasseri is difficult to grow in milk, which hurts usability for the food industry. It had been previously reported that supplementation with yeast extract or proteose peptone, including peptides, enables L. gasseri to grow well in milk. In this study, our objective was to confirm peptide requirement of L. gasseri and evaluate efficacy of peptide release by enzymatic proteolysis on growth of L. gassei in milk. Three strains of L. gasseri did not grow well in modified DeMan, Rogosa, Sharpe broth without any nitrogen sources (MRS-N), but addition of a casein-derived peptide mixture, tryptone, promoted growth. In contrast, little effect was observed after adding casein or a casein-derived amino acid mixture, casamino acids. These results indicate that L. gasseri requires peptides, not proteins or free amino acids, among milk-derived nitrogen sources for growth. Lactobacillus gasseri JCM 1131T hardly had growth capacity in 6 kinds of milk-based media: bovine milk, human milk, skim milk, cheese whey, modified MRS-N (MRSL-N) supplemented with acid whey, and MRSL-N supplemented with casein. Moreover, treatment with digestive proteases, particularly pepsin, to release peptides made it grow well in each milk-based medium. The pepsin treatment was the most effective for growth of strain JCM 1131T in skim milk among the tested food-grade proteases such as trypsin, α-chymotrypsin, calf rennet, ficin, bromelain, and papain. As well as strain JCM 1131T, pepsinolysis of milk improved growth of other L. gasseri strains and some strains of enteric lactobacilli such as Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus johnsonii, and Lactobacillus reuteri. These results suggest that some relatives of L. gasseri also use peptides as desirable nitrogen sources, and that milk may be a good supplier of nutritious peptides to enteric lactobacilli including L. gasseri after peptic digestion in the gastrointestinal tract. This is the first report showing peptide requirement of L. gasseri and efficacy of pepsinolysis on the growth of L. gasseri and its relatives in milk. This study would contribute to increasing usability of L. gasseri and its relatives as probiotics in dairy foods.
Topics: Amino Acids; Animals; Caseins; Cattle; Chymotrypsin; Gastrointestinal Tract; Lactobacillus; Milk; Nitrogen; Pepsin A; Peptide Fragments; Peptide Hydrolases; Peptides; Probiotics
PubMed: 25529420
DOI: 10.3168/jds.2014-8860 -
Applied and Environmental Microbiology Nov 2005Thirty-eight isolates of Lactobacillus gallinarum cultured from the crops of broiler chickens were screened for the presence of genes encoding S-layer proteins. All of...
Thirty-eight isolates of Lactobacillus gallinarum cultured from the crops of broiler chickens were screened for the presence of genes encoding S-layer proteins. All of the isolates had two S-protein genes, which were designated Lactobacillus gallinarum S-protein (lgs) genes. One gene in each isolate was either lgsA or lgsB. The Lactobacillus isolates were further characterized by pulsed-field gel electrophoresis of DNA digests, which grouped the isolates into 17 genotypes (strains). The second gene in each of eight representative strains was sequenced and shown to differ among strains (lgsC, lgsD, lgsE, lgsF, lgsG, lgsH, and lgsI). The genome of each strain thus encoded a common S-protein (encoded by either lgsA or lgsB) and a strain-specific S-protein. The extraction of cell surface proteins from cultures of the eight strains showed that each strain produced a single S-protein that was always encoded by the strain-specific lgs gene. Two of the strains were used to inoculate chickens maintained in a protected environment which were Lactobacillus-free prior to inoculation. DNAs and RNAs extracted from the digesta of the chickens were used for PCR and reverse transcription-PCR, respectively, to demonstrate the presence and transcription of lgs genes in vivo. In both cases, only the strain-specific gene was transcribed. Both of the strains adhered to the crop epithelium, consistent with published data predicting that S-proteins of lactobacilli are adhesins. The results of this study provide a basis for the investigation of gene duplication and sequence variation as mechanisms by which bacterial strains of the same species can share the same habitat.
Topics: Animals; Bacterial Proteins; Chickens; Crop, Avian; DNA, Bacterial; Deoxyribonucleases, Type II Site-Specific; Electrophoresis, Gel, Pulsed-Field; Lactobacillus; Membrane Glycoproteins; Molecular Sequence Data; Polymerase Chain Reaction; Species Specificity
PubMed: 16269691
DOI: 10.1128/AEM.71.11.6633-6643.2005 -
Poultry Science Jun 2021The objective of this study was to compare the effects of graded inclusions of 2 phytase products and a mineral P source in broiler chickens using different response...
The objective of this study was to compare the effects of graded inclusions of 2 phytase products and a mineral P source in broiler chickens using different response traits, including ileum microbiota composition. Eleven experimental diets were used. These were a low-P basal diet and diets supplemented with increasing levels of dicalcium phosphate (DCP), Natuphos E 5000 G (NE), or Natuphos 5000 G (N). The performance traits, prececal P digestibility, and tibia and foot ash results were subjected to regression analysis and slope ratios were used to compare the supplements based on the measured evaluation traits. In the microbiota analysis, total nucleic acids were extracted and the 16S rRNA gene was targeted for use in the amplicon sequencing process. Phylogenetic analysis was performed using Mothur, followed by a multivariate statistical analysis. The various response traits caused different estimates of relative efficacy. The mean results of all the response traits showed that a 1.75-fold increase in the activity of N was needed to achieve the same response as NE and the variability among the detected traits ranged from 1.59 (prececally digestible P intake) to 1.91 (amount of tibia ash). The mean slope ratio between DCP and NE was 311 and varied between 208 (ADG) and 349 (foot ash concentration). The mean slope ratio for phytase N with DCP was 552 and varied from 357 (ADG) to 640 (tibia ash concentration). The ileum microbiota composition was not different among the diets. A similar composition was driven in the abundance of Lactobacillus crispatus, Lactobacillus salivarius, and Lactobacillus gallinarum. The results suggest that different response traits cause markedly different estimates of relative phytase efficacy.
Topics: 6-Phytase; Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Chickens; Diet; Dietary Supplements; Digestion; Ileum; Lactobacillus; Microbiota; Phylogeny; RNA, Ribosomal, 16S
PubMed: 33940282
DOI: 10.1016/j.psj.2021.101133 -
Applied and Environmental Microbiology Nov 2003The microflora of the crop was investigated throughout the broiler production period (0 to 42 days) using PCR combined with denaturing gradient gel electrophoresis...
Detection and identification of Lactobacillus species in crops of broilers of different ages by using PCR-denaturing gradient gel electrophoresis and amplified ribosomal DNA restriction analysis.
The microflora of the crop was investigated throughout the broiler production period (0 to 42 days) using PCR combined with denaturing gradient gel electrophoresis (PCR-DGGE) and selective bacteriological culture of lactobacilli followed by amplified ribosomal DNA restriction analysis (ARDRA). The birds were raised under conditions similar to those used in commercial broiler production. Lactobacilli predominated and attained populations of 10(8) to 10(9) CFU per gram of crop contents. Many of the lactobacilli present in the crop (61.9% of isolates) belonged to species of the Lactobacillus acidophilus group and could not be differentiated by PCR-DGGE. A rapid and simple ARDRA method was developed to distinguish between the members of the L. acidophilus group. HaeIII-ARDRA was used for preliminary identification of isolates in the L. acidophilus group and to identify Lactobacillus reuteri and Lactobacillus salivarius. MseI-ARDRA generated unique patterns for all species of the L. acidophilus group, identifying Lactobacillus crispatus, Lactobacillus johnsonii, and Lactobacillus gallinarum among crop isolates. The results of our study provide comprehensive knowledge of the Lactobacillus microflora in the crops of birds of different ages using nucleic acid-based methods of detection and identification based on current taxonomic criteria.
Topics: Aging; Animals; Bacterial Typing Techniques; Chickens; Colony Count, Microbial; Crop, Avian; DNA, Bacterial; DNA, Ribosomal; Electrophoresis; Lactobacillus; Lactobacillus acidophilus; Polymerase Chain Reaction; RNA, Ribosomal, 16S; Restriction Mapping; Species Specificity
PubMed: 14602636
DOI: 10.1128/AEM.69.11.6750-6757.2003