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The Science of the Total Environment Feb 2024Megasphaera hexanoica is a bacterial strain following the reverse β-oxidation pathway to synthesize caproate (CA) using lactate (LA) as an electron donor (ED) and...
Megasphaera hexanoica is a bacterial strain following the reverse β-oxidation pathway to synthesize caproate (CA) using lactate (LA) as an electron donor (ED) and acetate (AA) or butyrate (BA) as electron acceptors (EA). Differences in the type and concentration of EA lead to distinctions in product distribution and energy bifurcation of carbon fluxes in ED pathways, thereby affecting CA production. In this study, the effect of various ratios of AA, BA, and AA+BA as EA on carbon flux and CA specific titer during the carbon chain elongation in M. hexanoica was explored. The results indicated that the maximum levels of CA were 18.81 mM and 31.48 mM when the molar ratios of LA/AA and LA/BA were 10:1 and 3:1, respectively. Meanwhile, when AA and BA were used as combined EA (LA, AA, and BA molar amounts of 100, 23, and 77 mM), a maximum CA production of 39.45 mM was obtained. Further analysis revealed that the combined EA exhibited a CA production carbon flux of 49 % (4.3 % and 19.5 % higher compared to AA or BA, respectively) and a CA production specific titer of 45.24 mol (80.89 % and 58.51 % higher compared to AA or BA, respectively), indicating that the effective carbon utilization rate and CA production efficiency were greatly improved. Finally, a scaled-up experiment was conducted in a 1.2 L (working volume) automated bioreactor, implying high biomass (optical density at 600 nm or OD = 1.809) and a slight decrease in CA production (28.45 mM). A decrease in H production (4.11 g/m) and an increase in CO production (0.632 g/m) demonstrated the appropriate metabolic adaptation of M. hexanoica to environmental changes such as stirring shear.
Topics: Fermentation; Carbon; Electrons; Bioreactors; Butyrates; Acetates; Caproates; Megasphaera
PubMed: 38141983
DOI: 10.1016/j.scitotenv.2023.169509 -
Animal Nutrition (Zhongguo Xu Mu Shou... Mar 2024Skatole, a strong fecal odor substance, is generated through microbial degradation of tryptophan in the animal hindgut. It easily accumulates in adipose tissue and...
Skatole, a strong fecal odor substance, is generated through microbial degradation of tryptophan in the animal hindgut. It easily accumulates in adipose tissue and affects meat quality. In this study, the effect of mulberry leaf supplementation on skatole in finishing pigs was studied. In a 35-day trial, 20 finishing pigs (barrows and gilts) were fed with a basal diet or basal diet with 6% mulberry leaves. Growth performance of the pigs ( = 10) was automatically recorded by a performance-testing feeder system and 8 pigs in each treatment were slaughtered and sampled for the remaining tests. Skatole and short-chain fatty acids were detected using HPLC and gas chromatography, respectively. Fecal microbiota were analyzed using 16S rRNA gene sequencing. The metabolomics analysis of feces and serum was performed with UHPLC-MS/MS. The major cytochrome P450 (CYP) enzymes that catalyze skatole degradation in the liver were tested by using RT-PCR and Western blot. Effects of major bioactive compounds in mulberry leaves on the genes were verified in the hepatic cell line HepG2 in an in vitro test ( = 3). In finishing pigs, mulberry leaf supplementation had no significant effect on the average daily gain, average daily feed intake, and feed conversion ratio ( > 0.05), but reduced skatole levels in feces, serum, and backfat ( < 0.05), and increased acetic acid levels in feces ( = 0.027). Mulberry leaf supplementation decreased the relative abundance of the skatole-producing bacteria and ( < 0.05). Indole-3-acetic acid, the intermediate that is essential for skatole production, was significantly reduced in feces by mulberry leaf supplementation ( < 0.05) and was positively correlated with skatole content in feces ( = 0.004). In pigs treated with mulberry leaves, liver expression was increased ( < 0.05) and was negatively correlated with skatole content in backfat ( = 0.045). The in vitro test demonstrated that mulberry leaf polyphenols and polysaccharides could directly stimulate expression in hepatic cells. These findings suggest that mulberry leaf supplementation reduces skatole production and deposition in finishing pigs by regulating the gut microbiota and promoting skatole degradation in liver.
PubMed: 38131029
DOI: 10.1016/j.aninu.2023.05.018 -
Applied and Environmental Microbiology Jan 2024Butyrate, a physiologically active molecule, can be synthesized through metabolic interactions among colonic microorganisms. Previously, in a fermenting trial of human...
Butyrate, a physiologically active molecule, can be synthesized through metabolic interactions among colonic microorganisms. Previously, in a fermenting trial of human fecal microbiota, we observed that the butyrogenic effect positively correlated with the increasing population and an unidentified species. Therefore, we hypothesized that a cross-feeding phenomenon exists between and , where is the butyrate producer, and its growth relies on the metabolites generated by . To validate this hypothesis, three bacterial species (, , and ) were isolated from fecal cultures fermenting hydrolyzed xylan; pairwise cocultures were conducted between the and isolates; the microbial interactions were determined based on bacterial genome information, cell growth, substrate consumption, metabolite quantification, and metatranscriptomics. The results indicated that two isolates contained distinct gene clusters for xylan utilization and expressed varying substrate preferences. In contrast, alone scarcely grew on the xylose-based substrates. The growth of was significantly elevated by coculturing it with bifidobacteria, while the two species responded differently in the kinetics of cell growth and substrate consumption. Coculturing led to the depletion of lactate and increased the formation of butyrate. An RNA-seq analysis further revealed the upregulation of genes involved in the lactate utilization and butyrate formation pathways. We concluded that lactate generated by through catabolizing xylose fueled the growth of and triggered the synthesis of butyrate. Our findings demonstrated a novel cross-feeding mechanism to generate butyrate in the human colon.IMPORTANCEButyrate is an important short-chain fatty acid that is produced in the human colon through microbial fermentation. Although many butyrate-producing bacteria exhibit a limited capacity to degrade nondigestible food materials, butyrate can be formed through cross-feeding microbial metabolites, such as acetate or lactate. Previously, the literature has explicated the butyrate-forming links between and and between and . In this study, we provided an alternative butyrate synthetic pathway through the interaction between and is a species named in 2014 and is indigenous to the human intestinal tract. Scientific studies explaining the function of in the human colon are still limited. Our results show that proliferated based on the lactate generated by bifidobacteria and produced butyrate as its end metabolic product. The pathways identified here may contribute to understanding butyrate formation in the gut microbiota.
Topics: Humans; Lactic Acid; Bifidobacterium; Xylans; Xylose; Butyrates; Megasphaera; Fermentation
PubMed: 38126785
DOI: 10.1128/aem.01019-23 -
Current Research in Microbial Sciences 2024Spontaneous fermentation of flour enhances the nutritional potential of this hybrid cereal. However, the effect of consumption of sourdough bread (SDB) on gut health...
Spontaneous fermentation of flour enhances the nutritional potential of this hybrid cereal. However, the effect of consumption of sourdough bread (SDB) on gut health remains to be elucidated. This study investigated the effect of digestion and faecal fermentation of SDB compared to that of traditional baker's yeast (BYB) bread. After 24-h anaerobic faecal fermentation, both SDB and BYB (1% w/v) induced an increase in the relative abundances of , and genera compared to baseline, while concentrations of acetate and butyrate were significantly higher at 24 h for SDB compared to those for BYB. Integrity of intestinal epithelium, as assessed through trans-epithelial electrical resistance (TEER) assay, was slightly increased after incubation with SDB fermentation supernatants, but not after incubation with BYB fermentation supernatants. The SDB stimulated mucosal immune response by inducing early secretion of inflammatory cytokines, IL-6 and TNF-α, followed by downregulation of the inflammatory trigger through induction of anti-inflammatory IL-10 expression. Overall, our findings suggest that sourdough can modulate gut microbiota fermentation activity and positively impact the gut health.
PubMed: 38116184
DOI: 10.1016/j.crmicr.2023.100214 -
Frontiers in Microbiology 2023Increasing evidence supports a role for the vaginal microbiome (VM) in the severity of HPV infection and its potential link to cervical intraepithelial neoplasia....
Increasing evidence supports a role for the vaginal microbiome (VM) in the severity of HPV infection and its potential link to cervical intraepithelial neoplasia. However, a lot remains unclear regarding the precise role of certain bacteria in the context of HPV positivity and persistence of infection. Here, using next generation sequencing (NGS), we comprehensively profiled the VM in a series of 877 women who tested positive for at least one high risk HPV (hrHPV) type with the COBAS 4,800 assay, after self-collection of a cervico-vaginal sample. Starting from gDNA, we PCR amplified the V3-V4 region of the bacterial 16S rRNA gene and applied a paired-end NGS protocol (Illumina). We report significant differences in the abundance of certain bacteria compared among different HPV-types, more particularly concerning species assigned to and genera. Especially for , we observed significant depletion in the case of HPV16, HPV18 versus hrHPVother. Overall, our results suggest that the presence or absence of specific cervicovaginal microbial genera may be linked to the observed severity in hrHPV infection, particularly in the case of HPV16, 18 types.
PubMed: 38098662
DOI: 10.3389/fmicb.2023.1292230 -
PloS One 2023Yeast-derived products have become more of an interest in the poultry industry as of late because of their use in modulating the gastrointestinal tract (GIT) microbiome... (Comparative Study)
Comparative Study
Yeast-derived products have become more of an interest in the poultry industry as of late because of their use in modulating the gastrointestinal tract (GIT) microbiome to both improve production parameters and prevent infection. This study aimed to evaluate the effects of various yeast-derived products on Salmonella enterica inoculation in un in vitro rooster cecal incubations and associated effects on the cecal microbiome. Cecal contents were obtained from 53-wk old White Leghorn H & N Nick Chick roosters (n = 3) fed a wheat-based, commercial-type basal diet. Cecal contents were diluted 1:3000 in anaerobic dilution solution (ADS) in an anaerobic chamber, with 20 mL aliquoted to each serum bottle. There were three controls (n = 3): basal diet only, diluted cecal contents only, and basal diet and diluted cecal contents; and five treatments containing the basal diet and diluted cecal contents (n = 3): Citristim® (ADM), ImmunoWall® (ICC), Maxi-Gen Plus® (CBS Bio Platforms), Hilyses® (ICC), and Original XPC® (Diamond V). All treatments were applied at a rate of 2.5 kg/tonne or less. All groups were inoculated with a nalidixic acid-resistant strain of Salmonella Enteritidis at 10^7 CFU/mL and incubated at 37 deg C. Samples were collected at 0, 24, and 48 h for S. Enteritidis enumeration and 16S rDNA microbial sequencing. Salmonella data were log-transformed and analyzed in a two-way ANOVA with means separated using Tukey's HSD (P≤0.05). Genomic DNA was extracted, and resulting libraries were prepared and sequenced using an Illumina MiSeq. Sequencing data were analyzed in QIIME2 (2021.4) with diversity metrics (alpha and beta), and an analysis of the composition of microbiomes (ANCOM) was performed. Main effects were considered significant at P≤0.05, with pairwise differences considered significant at Q≤0.05. There was an interaction of treatment and time on the enumeration of Salmonella where treatments of Citristim, Immunowall, Hilyses, and XPC reduced Salmonella by 1 log CFU/mL compared to the controls. At 48 h, each yeast product treatment reduced Salmonella by 3 log CFU/mL compared to the controls. There was no main effect of treatment on the alpha diversity metrics, richness, or evenness (P > 0.05). Treatment affected the beta diversity, abundance, and phylogenetic differences, but there were no pairwise differences (P>0.05, Q>0.05). Using ANCOM at the genus level, the taxa Synergistes, Alloprevotella, Sutterella, and Megasphaera abundance were significantly different (W = 154,147,145,140, respectively). These results demonstrate the potential of these yeast-derived products to reduce foodborne pathogens, such as Salmonella Enteriditis, in vitro, without negatively disrupting the cecal microbiome.
Topics: Animals; Male; Animal Feed; Cecum; Chickens; Diet; Microbiota; Phylogeny; Poultry Diseases; Saccharomyces cerevisiae; Salmonella enteritidis; Salmonella Infections, Animal; Gastrointestinal Microbiome
PubMed: 38096330
DOI: 10.1371/journal.pone.0295657 -
BMC Veterinary Research Dec 2023Acremonium terricola is used in the feed of dairy animals to promote growth and control diseases. However, the effects of dietary supplementation with A. terricola on...
Dietary supplementation with Acremonium terricola culture alters the gut microbial structure and improves the growth performance, antioxidant status, and immune function of weaning piglets.
BACKGROUND
Acremonium terricola is used in the feed of dairy animals to promote growth and control diseases. However, the effects of dietary supplementation with A. terricola on the gut microbial structure of weaning piglets remain poorly understood. Therefore, in this study, we investigated the effects of dietary supplementation with A. terricola culture (ATC) on the growth performance, antioxidant status, immunity, and gut environment of weaning piglets. Sixty piglets were fed a basal diet supplemented with 1 g ATC/kg of basal diet (experimental group). Another 60 piglets did not receive ATC (control group). The intervention lasted for 20 days.
RESULTS
The experimental group had higher daily weight gain and feed efficiency than did the control group. Significant increases were noted in the levels of serum insulin (P = 0.0018), insulin-like growth factor (P = 0.0018), triiodothyronine (P = 0.0031), immunoglobulin A (P < 0.0001), immunoglobulin M (P = 0.001), immunoglobulin G (P = 0.0001), and interferon γ (P < 0.0001) in the experimental group compared with the levels in the control group. Furthermore, ATC supplementation significantly reduced (P < 0.05) the relative abundance of Shuttleworthia, Succinivibrio, Roseburia, Ruminococcus, and Paludibacter but increased that of Phascolarctobacterium, Megasphaera, Faecalibacterium, and Prevotella in the experimental group compared with that in the control group. Notably, ATC supplementation significantly increased the relative abundance of Faecalibacterium prausnitzii (P < 0.05), which is involved in anti-inflammatory activities, gut barrier enhancement, and butyrate production.
CONCLUSIONS
Dietary supplementation with ATC may improve the growth performance, antioxidant status, immunity, and fecal microflora of weaning pigs.
Topics: Animals; Swine; Antioxidants; Weaning; Gastrointestinal Microbiome; Dietary Supplements; Diet; Immunity; Animal Feed
PubMed: 38053083
DOI: 10.1186/s12917-023-03778-y -
Bioresource Technology Feb 2024The objective was to investigate the impact of the bioaugmentation on chain elongation process using glycerol, lactate and lactose as substrates in an open culture...
The objective was to investigate the impact of the bioaugmentation on chain elongation process using glycerol, lactate and lactose as substrates in an open culture fermentation. In the batch trials the highest selectivity for chain elongation product, i.e. caproate, was observed in trials inoculated with co-culture of Megasphaera elsdenii and Eubacterium limosum grown on glycerol (28.6%), and in non-bioaugmented open culture run on lactose + lactate (14.8%). The results showed that E. limosum, out of two bioaugmented strains, was able to survive in the open culture. A continuous open culture fermentation of glycerol led to caproate and 1,3-propanediol (1,3-PDO) formation, while lactate addition led to 1,3-PDO and short chain carboxylates production. Moving the process into batch mode triggered even-carbon chain elongation. Presence of E. limosum promoted odd-carbon chain elongation and valerate production. Imaging flow cytometry combined with machine learning enabled the discrimination of Eubacterium cells from other microbial strains during the process.
Topics: Lactic Acid; Caproates; Carboxylic Acids; Glycerol; Lactose; Fermentation; Propylene Glycol; Carbon; Propylene Glycols
PubMed: 38042435
DOI: 10.1016/j.biortech.2023.130123 -
Animal : An International Journal of... Dec 2023Weaning is one of the most critical phases in pig's life, often leading to postweaning diarrhoea (PWD). Zinc oxide (ZnO), at pharmacological doses, has been largely used...
Weaning is one of the most critical phases in pig's life, often leading to postweaning diarrhoea (PWD). Zinc oxide (ZnO), at pharmacological doses, has been largely used to prevent PWD; however, due to antimicrobial co-resistant and environmental pollution issues, the EU banned its use in June 2022. Natural or natural identical components of essential oils and their mixture with organic acids are possible alternatives studied for their antimicrobial, anti-inflammatory and antioxidant abilities. This study aimed to evaluate the effect of two blends of natural or natural identical components of essential oils and organic acids compared to ZnO on health, performance, and gut health of weaned pigs. At weaning (d0), 96 piglets (7 058 ± 895 g) were assigned to one of four treatments balanced for BW and litter: CO (control treatment), ZnO (2 400 mg/kg ZnO from d0 to d14); Blend1 (cinnamaldehyde, ajowan and clove essential oils, 1 500 mg/kg feed); Blend2 (cinnamaldehyde, eugenol and short- and medium-chain fatty acids, 2 000 mg/kg feed). Pigs were weighed weekly until d35. Faeces were collected at d13 and d35 for microbiota (v3-v4 regions of the 16 s rRNA gene) and Escherichia coli (E. coli) count analysis. At d14 and d35, eight pigs/treatment were slaughtered; pH was recorded on intestinal contents and jejunal samples were collected for morphological and gene expression analysis. From d7-d14, the Blend2 had a lower average daily gain (ADG) than CO and ZnO (P < 0.05). ZnO and Blend1 never differed in ADG and feed intake. At d14, ZnO had a lower caecum pH than all other treatments. The CO treatment had a higher abundance of haemolytic E. coli than Blend1 (P = 0.01). At d13, the ZnO treatment had a lower alpha diversity (P < 0.01) and a different microbial beta diversity (P < 0.001) compared to the other treatments. At d13, the ZnO treatment was characterised by a higher abundance of Prevotellaceae_NK3B31_group (Linear Discriminant Analysis (LDA) score = 4.5, P = 0.011), Parabacteroides (LDA score = 4.5, P adj. = 0.005), the CO was characterised by Oscillospiraceae UCG-005 (LDA score = 4.3, P adj. = 0.005), Oscillospiraceae NK4A214_group (LDA score = 4.2, P adj. = 0.02), the Blend2 was characterised by Megasphaera (LDA score = 4.1, P adj. = 0.045), and Ruminococcus (LDA score = 3.9, P adj. = 0.015) and the Blend1 was characterised by Christensenellaceae_R-7_group (LDA score = 4.6, P adj. < 0.001) and Treponema (LDA score = 4.5, P adj. < 0.001). In conclusion, Blend1 allowed to maintain the gut health of postweaning piglets through modulation of the gut microbiome, the reduction of haemolytic E. coli while Blend2 did not help piglets.
Topics: Animals; Swine; Diet; Oils, Volatile; Escherichia coli; Zinc Oxide; Weaning; Diarrhea; Anti-Infective Agents; Dietary Supplements; Animal Feed
PubMed: 38035660
DOI: 10.1016/j.animal.2023.101031 -
Frontiers in Microbiology 2023Understanding the relationships between social stress and the gastrointestinal microbiota, and how they influence host health and performance is expected to have many...
Understanding the relationships between social stress and the gastrointestinal microbiota, and how they influence host health and performance is expected to have many scientific and commercial implementations in different species, including identification and improvement of challenges to animal welfare and health. In particular, the study of the stress impact on the gastrointestinal microbiota of pigs may be of interest as a model for human health. A porcine stress model based on repeated regrouping and reduced space allowance during the last 4 weeks of the finishing period was developed to identify stress-induced changes in the gut microbiome composition. The application of the porcine stress model resulted in a significant increase in salivary cortisol concentration over the course of the trial and decreased growth performance and appetite. The applied social stress resulted in 32 bacteria being either enriched (13) or depleted (19) in the intestine and feces. Fecal samples showed a greater number of microbial genera influenced by stress than caecum or colon samples. Our trial revealed that the opportunistic pathogens and were enriched in colonic and fecal samples from stressed pigs. Additionally, genera such as , , , , , and were found to be enriched in response to social stress. In contrast, the genera , , , , , , and were depleted. These depleted bacteria are of great interest because they synthesize metabolites [e.g., short-chain fatty acids (SCFA), in particular, butyrate] showing beneficial health benefits due to inhibitory effects on pathogenic bacteria in different animal species. Of particular interest are and , as their depletion was identified in a human study to be associated with inferior quality of life and depression. We also revealed that some pigs were more susceptible to pathogens as indicated by large enrichments of opportunistic pathogens of and . Generally, our results provide further evidence for the microbiota-gut-brain axis as indicated by an increase in cortisol concentration due to social stress regulated by the hypothalamic-pituitary-adrenal axis, and a change in microbiota composition, particularly of bacteria known to be associated with pathogenicity and mental health diseases.
PubMed: 38029169
DOI: 10.3389/fmicb.2023.1197371