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Validation of lactose[15N,15N]ureide as a tool to study colonic nitrogen metabolism.

In vitro experiments have shown that fermentation of carbohydrates prevents accumulation of nitrogen in the colon. Variable results have been obtained on modulation of dietary intakes in vivo. Lactose[15N,15N]-labeled ureide has been proposed as a tool to study colonic nitrogen metabolism. However, on oral administration of the marker, different urinary excretion patterns of the 15N label have been found. In this study, 50 mg lactose[15N,15N]ureide was directly instilled in the colon through an orocecal tube to investigate the colonic handling of this molecule in a direct way. In basal conditions, 42% (range, 37-48%) of labeled nitrogen administered as lactose[15N,15N]ureide was retrieved in urine after 72 h. A substantial variability in total urinary excretion of the label was found, but the urinary excretion pattern of the label was similar in all volunteers. When inulin, a fermentable carbohydrate, was administered together with the labeled marker, a significant decrease in urinary excretion of 15N after 72 h was found, to 29% (range, 23-34%). The effect of a smaller dose of inulin (250 mg) on colonic handling of lactose[15N,15N]ureide (50 mg), was investigated in another group of volunteers, and this time, fecal excretion of the marker was also evaluated. The results seem to indicate that fermentation of inulin causes an increased fecal excretion of the marker, thereby reducing urinary excretion but not retention in the human nitrogen pool. This instillation study shows that lactose[15N,15N]ureide is a tool with good properties to investigate the effect of different types of carbohydrates on nitrogen metabolism in the proximal colon in vivo.

Adult↗

Evaluation of the rice breath hydrogen test for small intestinal bacterial overgrowth.

OBJECTIVES: The aims of this study were 1) to document the sensitivity, specificity, and predictive values of the rice breath hydrogen test for small intestinal bacterial overgrowth; 2) to determine the possible influence of concurrent gastric bacterial overgrowth and gastroduodenal pH on the efficacy of this test; and 3) to investigate whether reliability is limited by an inability of small intestinal luminal flora to ferment rice or its product of hydrolysis, maltose. METHODS: Twenty adult subjects were investigated with microbiological culture of proximal small intestinal aspirate and a 3-g/kg rice breath hydrogen test. Gastroduodenal pH, the presence or absence of gastric bacterial overgrowth, and the in vitro capability of small intestinal luminal flora to ferment rice and maltose, its product of hydrolysis, were determined. RESULTS: Sensitivity of the rice breath hydrogen test for small intestinal bacterial overgrowth was 33% and remained low even when subjects with small intestinal overgrowth with oropharyngeal-type (38%) and colonic-type flora (20%) and those with concurrent small intestinal and gastric bacterial overgrowth (40%) were considered separately. Sensitivity remained suboptimal despite favorable gastroduodenal luminal pH and documented ability of bacterial isolates to ferment rice and maltose in vitro. Specificity of the rice breath hydrogen test for small intestinal bacterial overgrowth was 91%. Positive predictive value, negative predictive value, and predictive accuracy were 75%, 63%, and 65%, respectively. CONCLUSIONS: Clinical value of the rice breath hydrogen test for detecting small intestinal bacterial overgrowth is limited. The rice breath hydrogen test is not a suitable alternative to small intestinal intubation and culture of secretions for the detection of small intestinal bacterial overgrowth.

Adolescent↗

Fermented milks, probiotic cultures, and colon cancer.

Colorectal cancer (CRC) is a major cause of death from cancer in the Western world. Approximately 70% of CRC is associated with environmental factors, probably mainly the diet. There is interest in the potential protective role of fermented milks containing probiotic cultures against CRC. This article analyzes the existing data from human, animal, and in vitro studies that explore whether consumption of milks fermented with probiotic cultures could play a role in colon cancer risk reduction. Cohort studies have failed to detect significant effects, but most case-control studies favor a protective role of fermented milks against colon cancer. Interventional studies have shown a shift of intermediate markers of CRC risk in human subjects from a high- to low-risk pattern after ingestion of fermented milks or probiotics. Animal studies consistently show a reduction in chemically induced colorectal tumor incidence and aberrant crypt formation accompanying probiotic administration. In vitro studies also provide evidence of protection, and permit a better understanding of active compounds involved, and of the mechanisms underlying their anticarcinogenic effects. Probiotics may beneficially modulate several major intestinal functions: detoxification, colonic fermentation, transit, and immune status, which may accompany the development of colon cancer. Although the evidence is not conclusive and much further research is required, the data reviewed constitute a promising body of evidence supporting the protective role of milks fermented with probiotic cultures in colon cancer risk reduction.

Animals↗

Growth of Chaetomium cellulolyticum on Alkali-Pretreated Hardwood Sawdust Solids and Pretreatment Liquor.

The treatment of a hardwood sawdust with 1% NaOH solution at 121 degrees C dissolved 19.7% of the dry matter, mainly hemicellulose and lignin. Fermentation of the treated solids by Chaetomium cellulolyticum for 48 h gave a product containing 12.5% crude protein (total N x 6.25) on a dry weight basis. The in vitro rumen digestibility of the 48-h fermentation product was 30%, compared to 24% for the alkali-treated but unfermented sawdust. Growth was independent of sawdust particle size in the range 40 to 100 mesh. Fermentation of the pretreatment liquor gave a product containing up to 50% crude protein (dry weight basis) with an in vitro rumen digestibility of 65 to 76%. Approximately 6.7 g of crude protein was obtained from the treated solids and 2.2 g from the pretreatment liquor per 100 g of sawdust treated. The product from the pretreatment liquor fermentation has potential as a high-protein animal feed supplement but could not be produced economically without an outlet for the relatively indigestible product from the solids fermentation. Growth on the pretreatment liquor was strongly pH dependent; there was a considerable increase in the lag phase when the pH was lowered from 7.5 to 5.2. This effect appears to be due to an inhibitor whose toxicity is reduced at high pH.

Journal Article↗

Favourable effect of regular intake of fermented milk containing Lactobacillus johnsonii on Helicobacter pylori associated gastritis.

BACKGROUND: Lactobacillus johnsonii (Lj1) had an in vitro and in vivo inhibitory effect on Helicobacter pylori. Fermented milk containing Lj1 (LC1), coadministered with antibiotics had a favourable effect on H. pylori gastritis. AIM: Evaluate the effect of LC1 intake without antibiotics on H. pylori gastritis. METHODS: Fifty H. pylori positive healthy volunteers were randomised in a double-blind study to LC1 or placebo. Gastric biopsies from the antrum and corpus were obtained before, and after 3 and 16 weeks of treatment, for histology and quantitative cultures. RESULTS: Severity and activity of antral gastritis was reduced after 16-week LC1 intake (pretreatment and 16-week inflammatory cell score: 6.0 +/- 0.8 vs. 5.3 +/- 0.1; P=0.04). H. pylori density decreased in the antrum after LC1 intake (3-week: 4.4 +/- 0.6; 16-week: 4.3 +/- 0.5 log10 colony forming units (cfu) vs. pretreatment 4.5 +/- 0.4 log10 cfu; P=0.04, respectively). Mucus thickness increased after 16 weeks of LC1 consumption (change of mucus thickness with LC1 and placebo in the antrum: 0.6 +/- 1.3 vs. -0.2 +/- 1.0, P=0.01; in the corpus: 0.3 +/- 1.1 vs. -0.6 +/- 1.5, P=0.03). CONCLUSION: LC1 intake had a favourable, albeit weak, effect on H. pylori associated gastritis, particularly in the antrum. Regular ingestion of fermented milk containing L. johnsonii may reduce the risk of developing disorders associated with high degrees of gastric inflammation and mucus depletion.

Adolescent↗

Suppression by Hydrangeae Dulcis Folium of D-galactosamine-induced liver injury in vitro and in vivo.

Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla SER. var. thunbergii MAKINO, suppressed D-galactosamine-induced liver injury by 85.2% when added to the diet at 1% and fed to rats for fifteen days. The hepatoprotective effect is more potent than that of a milk thistle extract and turmeric powder. Some fractionated extracts showed hepatoprotective activity in the D-galactosamine-induced in vitro liver injury model.

Alanine Transaminase↗

[In vitro utilization of fructooligosaccharide by streptococci mutans].

Neosugar is the trade name of a fructooligosaccharide (FOS) whose utilization by oral bacteria is not well known yet. The aim of the present study was to evaluate in vitro the effect of this product on the growth, fermentation and production of plaque by mutans streptococci: S. mutans, serotypes c, e and f, S. sobrinus, serotype d, S. downei, serotype h, S. cricetus, serotype a and S. rattus, serotype b. The evaluation of growth was carried out in Brain Heart Infusion (BHI) broths containing or not sucrose and FOS and in buffered broths having glucose or FOS as carbon sources, through optical density reading in spectrophotometer after 24 hours of incubation at 37 degrees C. Thereafter the reading of pH was made in the same media. The plaque produced on glass sticks in BHI broths containing 5% sucrose or FOS was weighed and carbohydrates and proteins were assayed. The possible cariogenicity of Neosugar was confirmed, since it sustained the same growth and intensity of fermentation of sucrose in BHI broth for all streptococci and permitted in vitro production of plaque by some of them. The amount of plaque as well as its content of proteins and carbohydrates were smaller than those produced with sucrose, although the difference was statistically significant only for carbohydrates.

Animals↗

Butyrate is only one of several growth inhibitors produced during gut flora-mediated fermentation of dietary fibre sources.

Dietary fibre sources are fermented by the gut flora to yield short-chain fatty acids (SCFA) together with degraded phytochemicals and plant nutrients. Butyrate, a major SCFA, is potentially chemoprotective by suppressing the growth of tumour cells and enhancing their differentiation. Conversely, it could lead to a positive selection pressure for transformed cells by inducing glutathione S-transferases (GST) and enhancing chemoresistance. Virtually nothing is known about how butyrate's activities are affected by other fermentation products. To investigate such interactions, a variety of dietary fibre sources was fermented with human faecal slurries in vitro, analysed for SCFA, and corresponding SCFA mixtures were prepared. HT29 colon tumour cells were treated for 72 h with individual SCFA or complex samples. The growth of cells, GST activity, and chemoresistance towards 4-hydroxynonenal were determined. Fermentation products inhibited cell growth more than the corresponding SCFA mixtures, and the SCFA mixtures were more active than butyrate, probably due to phytoprotectants and to propionate, respectively, which also inhibit cell growth. Only butyrate induced GST, whereas chemoresistance was caused by selected SCFA mixtures, but not by all corresponding fermentation samples. In summary, fermentation supernatant fractions contain compounds that: (1) enhance the anti-proliferative properties of butyrate (propionate, phytochemical fraction); (2) do not alter its capacity to induce GST; (3) prevent chemoresistance in tumour cells. It can be concluded that fermented dietary fibre sources are more potent inhibitors of tumour cell growth than butyrate alone, and also contain ingredients which counteract the undesired positive selection pressures that higher concentrations of butyrate induce in tumour cells.

Aldehydes↗

Effect of domestic processing on the cooking time, nutrients, antinutrients and in vitro protein digestibility of the African yambean (Sphenostylis stenocarpa).

The effects of processing (soaking, dehulling, fermentation and heat treatment) on the cooking time, protein, mineral, tannin, phytate and in vitro protein digestibility (IVPD) of the African yambean (AYB) were examined. The cooking time ranged from 90-155 minutes. Soaking reduced cooking time by about 50 percent. Soaking for 12 hours was the most appropriate to reduce cooking time, tannin and phytate levels. It improved in vitro protein digestibility (IVPD). Prolonged soaking (24 hours) decreased calcium (Ca) and iron (Fe) values by 19 percent and 35 percent, respectively. Dehulling showed that Ca, Fe, magnesium (Mg) and zinc (Zn) were concentrated in the seed coat of the AYB. The seeds soaked and dehulled retained Mg and Zn. Dehulling reduced tannin but had no significant effect on phytate and the IVPD of the AYB except for seeds soaked for 12 hours before dehulling. Soaking for 24 hours before dehulling significantly increased crude protein content by 16 percent (p < 0.05). Blanching and roasting increased the IVPD by 8-11 percent. Fermentation had no effect on the crude protein, Ca, Fe, Mg and Zn but significantly reduced phytate content of the AYB. Fermentation had no advantage over heat treatment with respect to improving the in vitro protein digestibility of the AYB.

Africa↗

In vitro antimicrobial activity of the new antibiotic vermisporin.

The antimicrobial activity of vermisporin, a new antibiotic produced by fermentation of the fungus Ophiobolus vermisporis, was tested in vitro. Vermisporin inhibited 90% of Bacteroides fragilis and other Bacteroides spp. at 1 microgram/ml (range 0.25-1 micrograms/ml). Clostridium perfringens were inhibited by 1 microgram/ml (range 0.25-2 micrograms/ml). Vermisporin inhibited 90% of Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus, at 0.5 micrograms/ml (range 0.12-0.5 micrograms/ml). Vermisporin MICs for group A, B, C, F and G streptococci were < 1 microgram/ml when tested in Haemophilus Test Medium but > or = 8 micrograms/ml in the presence of blood. Vermisporin MICs for Enterobacteriaceae, Pseudomonas aeruginosa and Haemophilus influenzae exceeded 64 micrograms/ml. Inhibited organisms had MBCs 16- to 32-fold above the MICs.

Alkaloids↗

Effect of fermentation on the starch digestibility, resistant starch and some physicochemical properties of sorghum flour.

The effect of natural fermentation of Tabat sorghum cultivar (Sorghum bicolor L. Moench) at 37 degrees C for up to 36 h on pH, titratable acidity, starch digestibility, resistant starch and total starch was studied. The pH of the fermenting dough decreased sharply with a concomitant increase in the titratable acidity. In vitro starch digestibility markedly increased as a result of fermentation, while resistant starch and total starch decreased. Results showed that iodine absorption capacity increased during fermentation. Fermented sorghum had more soluble starch and swelling power at 100 degrees C than at 85 degrees C.

Chemical Phenomena↗

Contemporary in vitro spectrum of activity summary for antimicrobial agents tested against 18569 strains non-fermentative Gram-negative bacilli isolated in the SENTRY Antimicrobial Surveillance Program (1997-2001).

The frequency of occurrence and antimicrobial susceptibility patterns of 18569 non-fermentative Gram-negative bacilli consecutively collected as part of the SENTRY Antimicrobial Surveillance Program were summarized. The isolates were tested by the broth microdilution method in three coordinator laboratories using common reagents and reference methodologies. The most frequently isolated pathogen was Pseudomonas aeruginosa (11968 isolates; 64.5%) followed by Acinetobacter spp. (3468 isolates; 18.7%) and Stenotrophomonas maltophilia (1488 isolates; 8.0%). The lowest resistance rates for P. aeruginosa documented were for amikacin (8%), meropenem (10%) and cefepime (10%), and all fluoroquinolones tested showed similar resistance rates (22-24%). The most active compounds against Acinetobacter spp. were the carbapenems, imipenem (11% resistance) and meropenem (12% resistance) followed by cefepime (31% resistance) and gatifloxacin (32% resistance). Very few compounds showed reasonable in vitro activity against S. maltophilia, with the most active antimicrobial agents being trimethoprim/sulphamethoxazole, gatifloxacin and levofloxacin (5-6% resistance). Resistance surveillance among these organisms remains necessary to guide empirical antimicrobial therapy, especially for these less frequently isolated and difficult to test pathogens.

Bacterial Infections↗

Alterations in Krebs cycle enzyme activities and carbohydrate catabolism in two strains of Trypanosoma brucei during in vitro differentiation of their bloodstream to procyclic stages.

A rapid switch from a fermentative to a primarily oxidative type of glucose utilization was observed during in vitro differentiation of Trypanosoma brucei STIB348 and EATRO1244 bloodstream to procyclic trypomastigotes. In accordance with previously published reports bloodstream populations produced pyruvate as the major end product of glucose catabolism, together with very small amounts of CO2, succinate and glycerol. During differentiation pyruvate excretion decreased within 48 h to the low levels produced by 28-day procyclic stages. Concomitant with the decline in pyruvate formation, acetate appeared as a new product and the rates of respiratory CO2 increased considerably. The amount of carbon released with these compounds could account for nearly all of the glucose carbon consumed. Rates of glucose utilization and formation of acetate and CO2 in cells differentiated for 48 h were essentially the same as those found in 28-day procyclics. Succinate and glycerol excretion remained low during the entire transformation process, and no significant difference in the pattern and quantities of end products were found between the two trypanosome strains. During trypanosome differentiation the changes in metabolism were associated with marked alterations in enzyme activity levels. Activities of the tricarboxylic acid (TCA) cycle enzymes citrate synthase, isocitrate dehydrogenase (NAD+), succinate dehydrogenase and fumarase were not detectable in bloodstream trypomastigotes but appeared upon differentiation for 24 h. An exception was citrate synthase whose activity was not demonstrable until 48 h postinoculation into culture. After 48 h the majority of the TCA cycle enzyme activities continued to increase steadily until day 28. Pyruvate kinase activity decreased in differentiating cells after 48 h to about 25% of the level found in bloodstream trypomastigotes.(ABSTRACT TRUNCATED AT 250 WORDS)

Aconitic Acid↗

Dose-dependent production of mammalian lignans in rats and in vitro from the purified precursor secoisolariciresinol diglycoside in flaxseed.

The mammalian lignans enterodiol (ED) and enterolactone (EL) produced from colonic bacterial action on dietary precursors have exhibited anticarcinogenic effects in vitro. The major lignan precursor in flaxseed (a rich source) has been identified as secoisolariciresinol diglycoside (SDG). The purpose of this study was to first isolate SDG and determine whether 1) SDG accounted for all the lignan production from flaxseed; 2) this production was dose-related; and 3) a relationship between in vitro production and in vivo urinary excretion existed. Extraction of flaxseed with dioxane:ethanol (1:1, v/v) followed by chromatographic separations yielded the purified SDG. Rats were fed a high fat diet without/with 2.5, 5 or 10 g/100g ground flaxseed or 1.1, 2.2 or 4.4 micromol SDG/d (equivalent to levels in the respective flaxseed diets) for 4 wk. In vitro lignan production was assessed by fermenting flaxseed or SDG for 24 h with human fecal inoculum. Urinary lignan excretion increased linearly with doses from 0-5% flaxseed and 0-2.2 micromol SDG/d followed by a plateau, indicating a threshold response. When all doses were considered, a curvilinear relationship was observed. A similar trend was seen in vitro for SDG, resulting in a high correlation between in vitro production and in vivo excretion of lignans (r = 0.990, P < 0.02). Thus in vivo response can be predicted with confidence based on in vitro results. Theoretical urinary ED + EL from the SDG present in flaxseed correlated with the actual excretion in flaxseed-fed animals (r = 0.655, P < 0.005). However, urinary ED + EL of SDG-fed rats was only 20% of levels of flaxseed-fed rats, indicating the presence of other precursors or incomplete conversion of SDG to ED and EL.

4-Butyrolactone↗

Effects of dietary protein-energy interrelationships on Holstein steer performance and ruminal bacterial fermentation in continuous culture.

In vivo and in vitro 3 x 2 factorial experiments were conducted concurrently to evaluate the incorporation of 0, 15, or 30% sugar beet pulp (SBP) as an energy source in diets fed to growing Holstein steers with either soybean meal (SBM) or alcohol-treated, defatted soybean flakes (ATSBF) as primary supplemental protein sources. Three groups of 42 Holstein steers each were fed six different diets from 54 kg initial BW to 320 kg in three experimental periods. There were no overall SBP level x protein source interactions (P greater than .05). Beet pulp level tended to decrease ADG (linear, P = .05) and increase feed/gain (linear, P less than .05) and DMI (quadratic, P less than .05). Each grower diet was used in a substrate for ruminal microbial metabolism in six dual-flow, continuous-culture fermenters. Organic matter and carbohydrate digestion were similar (P greater than .05) among diets. Increasing dietary levels of SBP caused a concomitant increase (P less than .05) in acetate and decrease (P less than .05) in butyrate and isobutyrate concentrations. Beet pulp level x protein source interactions (P less than .05) were observed for CP degradation, ammonia and nonammonia N, and dietary N flow. Crude protein degradation was higher (P less than .05) for the 0% SBP with SBM diet (81.3%) than for the 30% SBP with ATSBF diet (64.4%). Efficiency of bacterial synthesis was similar (P greater than .05) among diets. Results indicated that SBP is an effective dietary energy source for high-energy grower diets at 15 or 30% of the DM but may cause a decrease in some performance traits. There were no nutritional benefits of using ATSBF vs SBM as the supplemental N source.

Animal Feed↗

In vitro gas production measurements to evaluate interactions between untreated and chemically treated rice straws, grass hay, and mulberry leaves.

In vitro gas production was measured to investigate associative effects of untreated and chemically treated rice straw (RS) and of RS and grass hay or mulberry leaves (ML). The RS was treated with NaOH, urea, or (NH4)HCO3. Cumulative gas production was recorded at 2, 4, 6, 8, 12, 16, 24, 36, 48, 60, 72, and 96 h of incubation, and the Gompertz function was used to describe the kinetics of gas production. Treatment with NaOH, urea, and NH4HCO3 increased (P < 0.05) gas production at 48 h by 55, 52, and 37% and the maximum rate of gas production of RS from 0.64 to 1.51, 1.27, and 1.13 mL/h, respectively. The inclusion of treated straws, hay, and ML in a mixture with RS at increasing proportions (25, 50, 75, 100%) elevated cumulative gas production and its rate. Maximum gas production was not different between the RS and its mixtures with the treated straws, but inclusion of hay into RS or NH4HCO3-treated RS increased the maximum gas production. Associative effects were defined as the difference between the observed gas production for the mixtures and the sum of the individual component feeds. The NaOH-treated RS, included at higher proportions, had positive associative effects at all times of incubation < 96 h; effects were negative or absent at 96 h. No effects were observed with mixtures of RS with urea-treated RS. The NH4HCO3-treated RS mixture tended to have negative associative effects at all three levels. Positive associative effects (P < 0.05) were observed for both RS and NH4HCO3-treated RS at almost all inclusion levels of hay or ML after 12 to 96 h of incubation. The response was more pronounced with ML than with hay. Associative effects generally declined with duration of incubation. We conclude that positive associative effects on in vitro gas production occurred more consistently when RS was incubated in mixtures with hay or ML than when incubated in mixtures with chemically treated RS.

Animals↗