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Human gut microbiota does not ferment erythritol.

Erythritol, a naturally occurring polyol, is gaining attention as a bulk sweetener for human nutrition. Industrially, it is produced from glucose by fermentation. From various studies it is known to be non-cariogenic. Moreover, it is rapidly absorbed in the small intestine and quantitatively excreted in the urine. Only about 10 % enters the colon. Earlier in vitro experiments showed that erythritol remained unfermented for a fermentation period of 12 h. In order to investigate whether fresh human intestinal microbiota is able to adapt its enzyme activities to erythritol, a 24 h lasting fermentation was carried out under well-standardised in vitro conditions. For comparison maltitol, lactulose and blank (faecal inoculum only) were incubated as well. Fermentation patterns were established by following total gas production, hydrogen accumulation, changes in pH value, SCFA production and substrate degradation. Taking all fermentation parameters into account, erythritol turned out to be completely resistant to bacterial attack within 24 h, thus excluding an adaptation within that period. Since under in vivo conditions more easily fermentable substrates enter the colon continuously, it seems very unlikely that erythritol will be fermented in vivo.

Erythritol↗

Immunomodulation of human blood cells following the ingestion of lactic acid bacteria.

Because of the lack of data that convincingly show immunomodulatory properties of lactic acid bacteria in humans, a study was performed in which healthy volunteers were divided into two groups and given a fermented milk product supplemented with Lactobacillus acidophilus strain La1 or Bifidobacterium bifidum strain Bb 12 for 3 wk. Blood was sampled throughout the study to assess changes in lymphocyte subsets or leukocyte phagocytic activity following consumption of the fermented products. No modifications of lymphocyte subpopulations were detected. In contrast, phagocytosis of Escherichia coli sp. in vitro was enhanced after the administration of both fermented products. The increment in phagocytosis was coincident with fecal colonization by the lactic acid bacteria and persisted for 6 wk after ingestion of the fermented products. By this time, the fecal lactobacilli and bifidobacteria had returned to concentrations prior to consumption. Nonspecific, anti-infective mechanisms of defense can be enhanced by the ingestion of specific lactic acid bacteria strains. These strains can be used as nutritional supplements to improve the immune function of particular age groups, i.e., the neonate or the elderly, for which these functions are diminished.

Adult↗

Biocontrol effect of a solid-state fermentation-derived extract mixture of Trichoderma asperellum on sunflower Sclerotinia rot and associated host defense responses.

Sclerotinia disease is a destructive fungal disease of sunflowers, soybeans, and other economically important crops, causing substantial yield loss and quality deterioration. Long-term reliance on dose-dependent broad-spectrum fungicides is constrained by resistance risks and potential environmental burdens, creating tension with the sustainability goal of "reducing pesticide use while improving efficacy." Here, we explore a Trichoderma spp.-based microbial disease management strategy. Whole-genome sequencing of Trichoderma asperellum TCS007 isolated from Antarctic marine sediments, coupled with genome mining, predicted diverse biosynthetic gene clusters putatively associated with siderophores, polyketides, nonribosomal peptides, and terpenoids; the corresponding metabolites are not chemically confirmed and require further validation. Using a solid-state fermentation workflow, we prepared a fermentation-derived extract mixture (TCS007-SSF-Ex). In vitro assays showed dose-dependent inhibition of Sclerotinia sclerotiorum by TCS007-SSF-Ex (EC50 = 1.252 mg/L), and microscopy revealed cellular damage-consistent changes, including organelle disruption and plasmolysis. Pathogen transcriptomic and metabolism-related analyses indicated broad perturbations in organelle biogenesis and metabolic processes, with significant alterations in pathways associated with succinate, D-glucose, and phenylacetate; these results are consistent with growth inhibition and reduced pathogenicity, but specific molecular targets and causal links remain to be validated. In vivo, under certain application conditions, triple applications increased APX activity (+492.5%) and β-1,3-glucanase activity (+419.6%). Collectively, this work supports a "pathogen suppression-host defense induction" framework and facilitates subsequent identification of active components and mechanistic validation.IMPORTANCESclerotinia diseases cause recurrent and economically important losses in oilseed crops, while long-term fungicide use is constrained by resistance risks and environmental burdens. Trichoderma-based biocontrol is a promising complementary strategy, yet evidence supporting metabolite-containing Trichoderma-derived preparations as immune elicitors remains less consolidated than that for living inoculants, and scalable production routes are still needed. Here, we examine an Antarctic marine sediment-derived strain, Trichoderma asperellum TCS007, and a solid-state fermentation (SSF)-derived extract mixture (TCS007-SSF-Ex) produced via solid-state fermentation. We combine in vitro antifungal assays, pathogen ultrastructural observations, and correlative omics analyses with in vivo measurements of sunflower defense enzymes (APX and β-1,3-glucanase) to evaluate a "pathogen suppression-host defense induction" framework. Our findings support the potential of SSF-derived Trichoderma metabolite mixtures for greener management of Sclerotinia disease and provide a foundation for future chemical identification of active components and mechanistic validation.

Ascomycota↗

BMS-182123, a fungal metabolite that inhibits the production of TNF-alpha by macrophages and monocytes.

A fungal metabolite, BMS-182123, which inhibited bacterial endotoxin-induced production of tumor necrosis factor (TNF-alpha) in murine macrophages and human peripheral blood monocytes (in vitro), was isolated from the culture broth of Penicillium chrysogenum strain V39673. The effective BMS-182123 concentration (IC50) resulting in 50% inhibition of lipopolysaccharide-induced TNF-alpha production in murine macrophages and human monocytes was 600 ng/ml and 4.0 microgram/ml, respectively. BMS-182123 suppressed the lipopolysaccharide-induced TNF-alpha promoter activity and did not affect the stability of posttranscriptional mRNA. Addition of hydrophobic resin, Amberlite XAD-8 (1%), to the fermentation enhanced the production of BMS-182123 by 5.5 fold. A total of 577 mg pure BMS-182123 was recovered from a 250-liter fermentation supplemented with 1% Amberlite XAD-8.

Animals↗

High density fermentation and activity of a recombinant lumbrokinase (PI239) from Pichia pastoris.

A system for the expression of recombinant lumbrokinase (rPI239) was developed in the yeast Pichia pastoris. A total supernatant protein content of 0.174 g/L of high density fermentation broth was obtained. The rPI239 exhibited in vitro fibrinolytic activity. The in vivo activity of rPI239 was measured by prothrombin time, kaolin part thrombin time, thrombin time, and fibrinolytic activity. This work presents the high-density fermentation of rPI239 from P. pastoris and shows that the recombinant protein has similar fibrinolytic activity both in vivo and in vitro.

DNA Primers↗

Effects of fermentation products of pro- and prebiotics on trans-epithelial electrical resistance in an in vitro model of the colon.

Evidence from in vivo and in vitro studies suggests that the consumption of pro- and prebiotics may inhibit colon carcinogenesis; however, the mechanisms involved have, thus far, proved elusive. There are some indications from animal studies that the effects are being exerted during the promotion stage of carcinogenesis. One feature of the promotion stage of colorectal cancer is the disruption of tight junctions, leading to a loss of integrity across the intestinal barrier. We have used the Caco-2 human adenocarcinoma cell line as a model for the intestinal epithelia. Trans-epithelial electrical resistance measurements indicate Caco-2 monolayer integrity, and we recorded changes to this integrity following exposure to the fermentation products of selected probiotics and prebiotics, in the form of nondigestible oligosaccharides (NDOs). Our results indicate that NDOs themselves exert varying, but generally minor, effects upon the strength of the tight junctions, whereas the fermentation products of probiotics and NDOs tend to raise tight junction integrity above that of the controls. This effect was bacterial species and oligosaccharide specific. Bifidobacterium Bb 12 was particularly effective, as were the fermentation products of Raftiline and Raftilose. We further investigated the ability of Raftilose fermentations to protect against the negative effects of deoxycholic acid (DCA) upon tight junction integrity. We found protection to be species dependent and dependent upon the presence of the fermentation products in the media at the same time as or after exposure to the DCA. Results suggest that the Raftilose fermentation products may prevent disruption of the intestinal epithelial barrier function during damage by tumor promoters.

Anticarcinogenic Agents↗

Stimulation of butyrate production by gluconic acid in batch culture of pig cecal digesta and identification of butyrate-producing bacteria.

Gluconic acid reaches the large intestine to stimulate lactic acid bacteria. However, the fermentation pattern of gluconic acid has yet to be elucidated. Accordingly, we examined the fermentation properties induced by gluconic acid in the pig cecal digesta in vitro. We also tested sorbitol and glucose, substrates for which the fermentation rate and patterns are known. The gluconic acid-utilizing bacteria were further isolated from pig cecal digesta and identified to examine the effect of gluconic acid on hind gut fermentation. Gluconic acid was fermented more slowly than were the other two substrates. Gluconic acid stimulated butyrate production; the butyrate molar percentage reached 26%, which is considered a high butyrate production. The majority of gluconic acid fermenters were identified as lactic acid bacteria, such as Lactobacillus reuteri and L. mucosae, and acid-utilizing bacteria, such as Megasphaera elsdenii and Mitsuokella multiacida. The gluconic acid fermented by lactic acid bacteria, and the lactate and acetate that were produced were used to form butyrate by acid-utilizing bacteria, such as M. elsdenii. Gluconic acid may be useful as a prebiotic to stimulate butyrate production in the large intestine.

Acetates↗

The endogenous polysaccharide utilization rate of mixed ruminal bacteria and the effect of energy starvation on ruminal fermentation rates.

When mixed ruminal bacteria were starved in vitro for 24 h, cellular ATP decreased, but there was little change in cell protein. Starved ruminal bacteria derived most of their ATP from cellular polysaccharide. Because polysaccharide declined at a first-order rate of 23%/h, it was possible to estimate the endogenous polysaccharide utilization rate at various stages of starvation by multiplying the amount of utilizable polysaccharide remaining at each time point by 0.23. The bacteria initially had a rate of soluble carbohydrate fermentation that was > 717 micrograms of hexose equivalent/mg of protein per h. Starvation had little impact on the rate of soluble carbohydrate fermentation until 8 to 12 h, and the endogenous polysaccharide utilization rate was < 10 micrograms of hexose/mg of protein per h. The bacteria digested ball-milled cellulose at a rate of 24 micrograms of hexose/mg of protein per h for 8 to 12 h. Even bacteria that had been starved for 24 h fermented cellulose at a rate of 16 micrograms of hexose/mg of protein per h. The rate of methane production was initially 70 nmol of methane/mg of protein per min. Short periods of starvation (< 12 h) had little impact on methane production, but longer times caused an almost complete inhibition of methanogenesis. The rate of amino acid deamination was initially 31 nmol of ammonia/mg of protein per min, and the critical phase of starvation was again 8 to 12 h. Ruminal bacteria that were harvested at 24 h after feeding had 10-fold less polysaccharide than did bacteria that were harvested at 2 h after feeding, but this polysaccharide supported high rates of soluble carbohydrate and cellulose fermentation, deamination, and methane production.

Adenosine Triphosphate↗

Non-lectin component in a fermented extract from Viscum album L. grown on pines induces proliferation of lymphocytes from healthy and allergic individuals in vitro.

Mistletoe preparations have been shown to express immunomodulatory properties. In order to evaluate the stimulatory potency of different mistletoe extracts, peripheral blood mononuclear cells (PBMC) from healthy and allergic/atopic individuals were exposed to aqueous or fermented extracts derived from Viscum album L. grown on apple trees (Mali-extracts) or on pines (Pini-extracts). None of them had received any mistletoe treatment. Iscador Pini was the only extract which strongly induced proliferation of PBMC in contrast to the other five preparations. On testing these extracts by Western blotting with anti-mistletoe lectin-1 (ML-1) antibody positive sera from mistletoe-treated patients, it became evident that Iscador Pini was almost devoid of lectins. The stimulatory potency of Iscador Pini for PBMC from three different groups was examined: PBMC from 35 normal controls (Group I), 23 patients with drug-induced adverse effects (Group II) and 16 individuals with allergic manifestations (Group III). Cells were exposed in 7-day cultures to the extract at concentrations between 1 and 10,000 micrograms/ml. PBMC from 63% of Group III individuals showed strong stimulation (SI varying from 6 to 97) in contrast to only 9% from Group I and 22% from Group II individuals. Anti-ML-1 antibodies were detected in 5% and anti-IP antibodies in 11% of subjects in the three groups. They were either of the IgA or IgM type but not of the IgG type. Our findings strongly imply that a non-lectin associated antigen from Iscador Pini is able to activate PBMC from healthy and allergic/atopic individuals, thereby demonstrating sensitization to probably highly conserved plant antigens.

Adult↗

Regulation of nitrogen fixation in Rhodospirillum rubrum grown under dark, fermentative conditions.

Rhodospirillum rubrum was shown to grow fermentatively on fructose with N2 as a nitrogen source. The nitrogenase activity of these cells was regulated by the NH4+ switch-off/switch-on mechanism in a manner identical to that for photosynthetically grown cells. In vitro, the inactive nitrogenase Fe protein from fermenting cells was reactivated by an endogenous membrane-bound, Mn2+-dependent activating enzyme that was interchangeable with the activating enzyme isolated from photosynthetic membranes.

Darkness↗

The fermentable fiber content of the diet alters the function and composition of canine gut associated lymphoid tissue.

The ingestion of plant fibers and their susceptibility to microbial fermentation in the large bowel modulate intestinal morphology but little is known about effects on the gut associated lymphoid tissue (GALT). The aim of the present study was to determine the effect of consuming diets containing different levels of fermentability fiber on immune function. Sixteen adult mongrel dogs (23 +/- 2 kg) were fed (14 days) in a randomized cross over design two isoenergetic isonitrogenous diets containing 8.3 g/kg non-fermentable or 8.7 g/kg fermentable fibers. Lymphocytes were isolated from blood prior to starting the study and at the end of each diet period. At study completion, lymphocytes were isolated from the gut associated lymphoid tissue (GALT) of the small intestine for characterization by immunofluorescence and to determine their ability to respond to mitogenic stimulation. Feeding high fermentable fibers increased (P < 0.05) the CD4/CD8 ratio and decreased (P < 0.05) the proportion of B cells in peripheral blood without changing natural killer cell activity or the response to mitogens. Mesenteric lymph node cells from dogs fed the low then high fermentable fiber diet contained a higher (P < 0.05) proportion of CD4+ cells and a higher (P < 0.05) response to mitogens. Intraepithelial, Peyer's patches and lamina propria cells contained a greater (P < 0.05) proportion of CD8+ cells when dogs were fed a low fermentable fiber diet followed by a high fermentable fiber diet. T cell mitogen responses in vitro were higher for intraepithelial but lower for Peyer's patches and lamina propria cells from dogs who were fed the low fermentable fiber diet followed by the high fermentable fiber diet (P < 0.05). In conclusion, the fermentable fiber content of the diet had very little effect on the type and function of immune cells in peripheral blood. However, feeding dogs a high fermentable fiber diet for 2 weeks (after 2 weeks of consuming a low fermentable fiber diet) altered the T-cell composition of GALT and produced a higher mitogen response in the predominantly T cell tissues and a lower response in areas involved in B cell functions. In conclusion, the level of fermentable fiber in the diet appears to alter GALT properties. Further studies are required to determine the direct contribution of a high or low fiber diet to these changes and the physiological implications to the health of the animal.

Animals↗

Lactic acid fermentation stimulated iron absorption by Caco-2 cells is associated with increased soluble iron content in carrot juice.

An in vitro digestion/Caco-2 cell model was applied to explore the impact of lactic acid (LA) fermentation by Lactobacillus pentosus FSC1 and Leuconostoc mesenteroides FSC2 on the Fe bioavailability of carrot juice. The redox state of Fe in fermented carrot juice was also assessed as a crucial factor for absorption. LA fermentation was shown to improve mineral solubility to different extents at simulated physiological conditions: Mn (2-fold); Fe (1.5-1.7-fold); Zn (1.2-fold); Cu (1-fold). Soluble Fe2+ was increased about 16-fold by LA fermentation, and about one third of the Fe2+ remained soluble after in vitro digestion (about 4-5-fold higher than in fresh juice). Data on cell-line studies showed a 4-fold increase in the efficiency of Fe uptake, but not in transepithelial transfer by Caco2 cells, as a result of fermentation. The increases in Fe2+ level and the efficiency of cellular Fe uptake were strain-dependent. To sum up the effect on both Fe solubility and cellular uptake efficiency, the amount of cellularly absorbed Fe from Ln. mesenteroides FSC2-fermented juice was about 20 % higher than that from L. pentosus FSC1-fermented juice (22.7 v. 19.2 microg/l juice per mg protein). To conclude, LA fermentation enhanced Fe absorption by Caco-2 cells from carrot juice because of increases in not only Fe solubility after digestion, but also the efficiency of cellular Fe uptake. The fermentation-improved efficiency of Fe uptake was possibly due to the increased level of soluble Fe2+ rather than a being a strain-specific event.

Beverages↗

Effect of thyme essential oils (Thymus hyemalis and Thymus zygis) and monensin on in vitro ruminal degradation and volatile fatty acid production.

The effect of the essential oils of thyme on the in vitro ruminal degradability of a barley seed/alfalfa hay substrate was studied. Two essential oils were used, one from Thymus hyemalis (TH), rich in carvacrol, and the other from Thymus zygis (TZ), rich in thymol. Four experimental treatments of in vitro degradability, using the Daisy II(200/220) incubator, were conducted including a negative control (CO), a positive control at 7.5 microg/mL of monensin (MO), and two treatments with essential oils (TH or TZ) at 1.35 microL/mL. The material was incubated at 39.5 degrees C for various lengths of time. At each time, the disappearance of dry matter, crude protein, and neutral detergent fiber was measured. Volatile fatty acids (VFAs) were determined after 48 h of incubation. CO and MO provided (p < 0.01) higher values of potential degradability (a + b) of DM than the TH and TZ treatments (72.6 and 70.8 vs 53.2 and 48.2%, respectively). Also, crude protein degradability was lowest in the essential oil treatments. The CO treatment showed the highest potential degradability of NDF. The values of VFA production obtained (p < 0.001) with CO and MO treatments were higher than those obtained with TH and TZ treatments (21.0 and 19.1 vs 11.2 and 10.1 mM). The essential oils decreased the molar proportion of propionate, increasing the acetate/propionate ratio. In conclusion, the effects of essential oils at assayed doses would not be nutritionally beneficial to the ruminal energetic metabolism.

Animals↗

Stimulation of bicarbonate secretion by luminal short-chain fatty acid in the rat and human colon in vitro.

Acetic, propionic and butyric acids constitute the predominant luminal anions in the colon. Several in vivo studies have suggested that these short-chain fatty acids (SCFAs) induce HCO3- accumulation in the colonic lumen. The purpose of this study was to delineate the mechanism underlying the SCFA-induced HCO3- accumulation in chamber-mounted mucosae from the rat and human colon under short-circuit conditions. HCO3- transport from the serosa bathed with a HCO3(-)-containing solution to the lumen bathed with a HCO3- (and Cl-)-free solution was evaluated from the luminal alkalization rate (JOHSL) by using a pH-stat system. In the rat distal colon, luminal propionate (25 mM) enhanced JOHSL from 0.32 +/- 0.19 to 3.18 +/- 0.22 mumol/cm2/h (n = 5, means +/- SE). Luminal acetate and butyrate (25 mM) similarly enhanced JOHSL. The magnitude of the increase in JOHSL induced by SCFA did not significantly vary among these three SCFAs. On the other hand, luminal lactate (25 mM) failed to affect JOHSL. The SCFA-induced increase in JOHSL was greatly reduced to approximately 30% of the original level when HCO3- was removed from the serosal solution. The short-circuit current and the transepithelial conductance were barely altered by luminal SCFAs or lactate. In the human colon, luminal propionate (25 mM) also caused an increase in JOHSL from 0.54 +/- 0.36 to 1.57 +/- 0.62 mumol/cm2/h (n = 4), which was significantly attenuated by removing serosal HCO3-. These results demonstrate that luminal SCFAs stimulate HCO3- secretion in the colon.

Acetates↗

Laidlomycin butyrate--an ionophore with enhanced intraruminal activity.

The polyether ionophore, laidlomycin, and several acyl derivatives were tested for their ability to favorably alter fermentation in two types of in vitro rumen fluid incubations. Dose response data were used to estimate the concentration (microgram/ml) of each ionophore required for either 50% maximal enhancement of propionic acid production (EC50) or 50% maximal inhibition of lactic acid production (IC50). Acylation of laidlomycin with straight-chain acyl groups from two to 12 carbon atoms tended to improve the potency of laidlomycin, especially for inhibiting lactic acid production. Comparative incubations using laidlomycin butyrate, laidlomycin and monensin indicated that both laidlomycin butyrate (EC50 = .3) and monensin (EC50 = .7) were more potent enhancers of propionic acid production than laidlomycin (EC50 = 2.0; P less than .05). Laidlomycin butyrate (IC50 = .3) was a more potent inhibitor of lactic acid production than either laidlomycin (IC50 = 1.8) or monensin (IC50 = 1.3; P less than .05). In a continuous culture experiment, three chemostats each received laidlomycin butyrate or monensin at the rate of .5 micrograms/ml of effluent/d while two flasks remained as control. Propionic acid production was increased (P less than .01) from 22.9 mmol/d in control flasks to 30.5 and 33.7 mmol/d in flasks treated with monensin and laidlomycin butyrate, respectively. Concomitant decreases in production rates of acetic, butyric and isovaleric acids also were observed (P less than .01). Thirty-six steers were used in a 56-d trial to evaluate effects of laidlomycin butyrate and monensin, at 33 mg/kg of diet, on feedlot performance.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Effect of cetylpyridinium chloride on formation and metabolism of human dental plaque].

In this work, the Cepacol (cetylpyridinium chlorid) diluted 1:2, when used for mouthwashes three time a day decreased the "in situ" formation of human dental plaque, however it didn't decreased neither the plaque fermentation, nor the IEP synthesis by the plaque. When the Cepacol was used for treating the "in vitro" dental plaque in both 1:10 and 1:20 dilutions, decreased the fermentation and the IEP synthesis of the "in vitro" plaque.

Cetylpyridinium↗