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Poultry Science Feb 2024Chickens in commercial production are hatched in hatcheries without any contact with their parents and colonization of their skin and respiratory tract is therefore...
Chickens in commercial production are hatched in hatcheries without any contact with their parents and colonization of their skin and respiratory tract is therefore dependent on environmental sources only. However, since chickens evolved to be hatched in nests, in this study we evaluated the importance of contact between hens and chicks for the development of chicken skin and tracheal microbiota. Sequencing of PCR amplified V3/V4 variable regions of the 16S rRNA gene showed that contact with adult hens decreased the abundance of E. coli, Proteus mirabilis and Clostridium perfringens both in skin and the trachea, and Acinetobacter johnsonii and Cutibacterium acnes in skin microbiota only. These species were replaced by Lactobacillus gallinarum, Lactobacillus aviarius, Limosilactobacillus reuteri, and Streptococcus pasterianus in the skin and tracheal microbiota of contact chicks. Lactobacilli can be therefore investigated for their probiotic effect in respiratory tract in the future. Skin and respiratory microbiota of contact chickens was also enriched for Phascolarctobacterium, Succinatimonas, Flavonifractor, Blautia, and [Ruminococcus] torque though, since these are strict anaerobes from the intestinal tract, it is likely that only DNA from nonviable cells was detected for these taxa.
Topics: Animals; Female; Chickens; RNA, Ribosomal, 16S; Escherichia coli; Microbiota; Respiratory System
PubMed: 38052128
DOI: 10.1016/j.psj.2023.103302 -
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 -
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 -
3 Biotech Dec 2022Lactic acid bacteria (LAB) are believed to have health-promoting properties to the host and can be used in therapeutics interventions; intriguingly, they have the...
UNLABELLED
Lactic acid bacteria (LAB) are believed to have health-promoting properties to the host and can be used in therapeutics interventions; intriguingly, they have the property to produce bio-preservatives substances. Therefore, this study aimed to mine probiotics and evaluate their safety, functional properties, and cholesterol-lowering capability. Seven potential probiotic strains were compared from 56 LAB strains isolated from traditional Chinese fermented milk. The results showed that all tested strains are tolerant to gastric acidity (45.5-83.26) and bile salts (11.92-92.91%) and have antibacterial activity against ATCC25923 and ATCC25922. Likewise, it lowered the cholesterol levels in vitro by live cells (26.57-45.76%) and dead cells (29.53-50.97%) with remarkable aggregation ability (13.8-43.71%). Antioxidant properties and produce short chain fatty acids (SCFAs) were strain-dependent features. Upon assessment of the safety, NWAFU-BIO-AS14 exhibited virulence factors genes (VFs) of (mur-2ed, odc, and tet(K)) and + hemolysis activity. While NWAFU-BIO-A-B24 and NWAFU-BIO-B-S6 have VFs of (odc, vanC2, and ant(6)-Ia). NWAFU-BIO-D-B2 has only (odc). Thus, they are not considered as safe probiotics. In contrast, NWAFU-BIO-BS29, NWAFU-BIO-AS16, and NWAFU-BIO-D-S7 are the safest and best strains, respectively, due to the absence of 16 VFs and their sensitivity to antibiotics such as kanamycin, erythromycin, tetracycline, gentamycin, vancomycin, streptomycin, chloramphenicol, and ampicillin. Accordingly, these strains have a high potentiality to be used as starter cultures or safely applied as perfect probiotics in functionals food and feed.
SUPPLEMENTARY INFORMATION
The online version contains supplementary material available at 10.1007/s13205-022-03403-z.
PubMed: 36340806
DOI: 10.1007/s13205-022-03403-z -
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 -
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 -
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 -
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 -
Journal of Food Science and Technology Mar 2018In this study, it was aimed to determine microbial flora members in three traditional Tulum cheeses (C1, C2 and C3) produced in different villages and settlement areas...
In this study, it was aimed to determine microbial flora members in three traditional Tulum cheeses (C1, C2 and C3) produced in different villages and settlement areas in İzmir, Turkey. For this purpose, culture depended and 16S rRNA based culture independent methods were used. According to the results of culture depended method, spp., spp., spp., spp., spp. and yeast-mold were detected in all samples at different levels. In order to determine and identify both of the culturable and non-culturable microorganisms, denaturing gradient gel electrophoresis (DGGE) method was used. DGGE results have shown that there were eight different dominant microorganisms (, subs. , subs. , , , , , ) in three regionally cheese samples. Further more, total bacterial loads were monitored with real-time PCR (qPCR) method. According to the results, 3.5 × 10, 3.8 × 10, 8.4 × 10 copy number of DNA was detected in C1, C2 and C3 cheese samples, respectively. This study is the first description for the dynamics of microbial composition of Izmir Tulum cheese after the production and brining processes.
PubMed: 29487437
DOI: 10.1007/s13197-017-3003-z -
Frontiers in Pharmacology 2016Food and Drug Administration (FDA, USA)-approved category B antibiotics are commonly prescribed to treat infections during pregnancy. The aim of this study was to...
Food and Drug Administration (FDA, USA)-approved category B antibiotics are commonly prescribed to treat infections during pregnancy. The aim of this study was to investigate antibiotic-induced changes in gut microbiota (GM) that occur during pregnancy. The 16S rRNA amplicon deep-sequencing method was used to analyze the effect of category B antibiotics (azithromycin, amoxicillin and cefaclor) on GM during pregnancy using a rat model. The GM composition was substantially modulated by pregnancy and antibiotics administration. Firmicutes, Bacteroidetes, Proteobacteria, Chlamydiae, Actinobacteria, and Cyanobacteria were the dominant phyla. Antibiotic treatment during pregnancy increased the relative abundance of Proteobacteria and reduced Firmicutes. The genera Shigella, Streptococcus, Candidatus Arthromitus, and Helicobacter were significantly (p < 0.05) more abundant during pregnancy. Antibiotics significantly (p < 0.05) reduced the relative abundance of Lactobacillus but increased that of Enterobacter. There was a significant (p < 0.05) decrease in Lactobacillus sp., Lactobacillus gallinarum and Lactobacillus crispatus during pregnancy. Antibiotic treatment reduced bacterial diversity; the lowest number of operational taxonomic units (OTUs) were detected in the cefaclor-treated groups. Antibiotics significantly (p < 0.05) promoted weight gain during pregnancy, and increased relative abundance of Shigella sonnei, Enterococcus hormaechei, and Acinetobacter sp. GM perturbations were accompanied by increases in Proteobacteria abundance and weight gain in pregnancy following antibiotic treatment.
PubMed: 27199748
DOI: 10.3389/fphar.2016.00104