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PubMed1999

Use of in vitro gas production models in ruminal kinetics.

Physiological systems models for ruminant animals are used to predict the extent of ruminal carbohydrate digestion, based on rates of intake, digestion, and passage to the lower tract. Digestion of feed carbohydrates is described in these models by a first-order rate constant. Recently, an in vitro gas production technique has been developed to determine the digestion kinetics in batch fermentation, and nonlinear mathematical models have been fitted to the cumulative gas production data from these experiments. In this paper, we present an analysis that converts these gas production models to an effective first-order rate constant that can be used directly in rumen systems models. The analysis considers the digestion of an incremental mass of substrate entering the rumen. The occurrence of passage is represented probabilistically, and integration through time gives the total mass of substrate and total rate of digestion in the rumen. To demonstrate the analysis, several gas production models are fitted to a sample data set for corn silage, and the effective first-order rate constants are calculated. The rate constants for digestion depend on ruminal passage rate, an interaction that arises from the nonlinearity of the gas production models.

Animals↗
PubMed1999

Mutagenicity of commercial Monascus fermentation products and the role of citrinin contamination.

Pigments produced as secondary metabolites by various isolates of moulds belonging to the genus Monascus have been used traditionally as colorants in Oriental food. Modern food industry has rediscovered these moulds as promising source for natural colorants. However, recent studies evidence that one of the secondary metabolites produced by Monascus is identical in structure to the mycotoxin citrinin. Thus, a sensitive HPLC method was developed to analyse these food colorants for contamination with citrinin. The mycotoxin could be detected in all the commercial Monascus samples at concentrations varying between 0.2 to 17.1 microg/g. In addition, the mutagenicity of commercial Monascus samples applying Salmonella-microsome assay and Salmonella-hepatocyte-assay was investigated and compared to the results obtained with citrinin. Citrinin and two Monascus extracts induced a positive dose depending mutagenic response in the Salmonella-hepatocyte-assay applying strain TA-98. However, no mutagenicity could be detected in the Salmonella-microsome assay, neither with nor without S9-mix, for citrinin and Monascus extracts, applying TA-98, TA-100, TA-1535, TA-1538 and TA-97. These findings provide further evidence that citrinin requires complex cellular biotransformation to exert mutagenicity.

Animals↗
PubMed1999

Oxidant stress: the role of nutrients in cell-lipoprotein interactions.

Oxidant stress is increasingly becoming an important hypothesis to explain the genesis of several pathologies, including cancer, atherosclerosis and also ageing. Beside a few rare genetic defects, dietary factors are thought to play a key role in the regulation of the production of reactive oxygenated species. An imbalance between nutrients, and in particular those involved in antioxidant status, could explain the onset of an enhanced production of free radicals. We will briefly review information concerning oxidation of lipids and lipoproteins which lead to atherothrombosis. We also present new findings supporting a role for blood platelets in generating oxidant species. New data are also described concerning the role of oxygenated derivatives of cholesterol, oxysterols, in cellular cholesterol efflux and NO production. Also, new developments relating to the influence of direct effects of free radicals on cellular cholesterol homeostasis are presented. Finally, the in vitro effects of butyrate, a natural short-chain fatty acid produced by bacterial fermentation, in the protection against free radical-mediated cytotoxicity are discussed. These data provide information on the mechanisms of dietary antioxidants in preventing oxidant stress.

Animals↗
PubMed1989

Yogurt and fermented-then-pasteurized milk: effects of short-term and long-term ingestion on lactose absorption and mucosal lactase activity in lactase-deficient subjects.

Lactase-deficient subjects absorb lactose in yogurt more effectively than lactose in other dairy products. However, as all previous studies were performed without a double-blind design and only after a single ingestion of the test product, the mechanism of this enhanced absorption remains unclear. The aims of this double-blind study were 1) to evaluate lactose absorption after prolonged ingestion of yogurt and fermented-then-pasteurized milk (FPM) and 2) to assess the modification of the lactase activity of the duodenal mucosa. In 16 lactase-deficient subjects we confirmed that yogurt enhances lactose digestion, this beneficial effect being destroyed by pasteurization. Moreover, the long-term (8 d) ingestion of either yogurt or FPM does not modify the results of hydrogen breath tests in comparison with a 24-h ingestion. The mucosal lactase (Dahlquist method) and beta-galactosidase (ONPG method) activities were not significantly modified by yogurt or FPM ingestion. These results suggest that in lactase-deficient subjects no adaptation occurs after eating yogurt or FPM and that the increased lactose absorption in yogurt must be mainly related to an intraluminal process.

Adolescent↗
PubMed1992

Metabolism and disposition of erythritol after oral administration to rats.

The metabolism and disposition of erythritol was studied using [14C]erythritol in rats. When [14C]erythritol was administered orally at a dose of 0.1 g/kg body wt to male rats, only 6% of the total radioactivity was excreted as expired 14CO2 and 88% was excreted in the urine within 24 h. The excreted metabolite in the urine consisted of a single component identified as intact [14C]erythritol. The excretion of 14CO2 and the incorporation ratios of radioactivity into tissues increased with the oral dosage. After rats were given an intravenous injection of [14C]erythritol, approximately 1% was excreted as 14CO2 and greater than 94% was excreted in the urine as intact [14C]erythritol. The excretion of 14CO2 within 24 h was increased to approximately 10% when [14C]erythritol was administered to rats that had been adapted to erythritol by feeding a diet containing 10% erythritol for 2 wk. When [14C]erythritol was incubated in vitro with the cecal contents from rats adapted to erythritol, greater than 20% was fermented to 14CO2 and 60% to short-chain fatty acids in 6 h. These results indicate that most orally administered erythritol was excreted in the urine without any degradation and that the remainder was transferred to the lower intestine and fermented by microbes.

Administration, Oral↗
PubMed2001

Interspecies H2 transfer in cellulose degradation between fibrolytic bacteria and H2-utilizing microorganisms from the human colon.

Interspecies H2 transfer between two newly isolated fibrolytic strains (18P13 and 18P16) and H2-utilizing methanogen or acetogen from the human colon was investigated during in vitro cellulose degradation. Both H2-consuming microorganisms utilized efficiently H2 produced from cellulose fermentation by the fibrolytic species. H2 utilization by Methanobrevibacter smithii did not change the metabolism and the cellulolytic activity of strain 18P16 whereas it induced a metabolic shift in strain 18P13. However, this metabolic shift was not associated with enhancement of cellulose degradation. In contrast, an increase in cellulose breakdown was observed when strain 18P13 was cultivated with Ruminococcus hydrogenotrophicus. This stimulating effect could be attributed to both the autotrophic and the heterotrophic metabolism of the acetogen in the coculture.

Anaerobiosis↗
PubMed1984

Derepression of citrate synthase in Saccharomyces cerevisiae may occur at the level of transcription.

Pulse-chase labeling in whole cells and cell-free protein synthesis were used to establish that the mitochondrial enzyme citrate synthase is made as a larger precursor in Saccharomyces cerevisiae. A 54,000 Mr precursor form appeared to be a primary translation product since it could be labeled with N-[35S]formylmethionine in vitro. The induction of citrate synthase was monitored in S. cerevisiae cells grown on fermentable (glucose) and nonfermentable (ethanol and glycerol) carbon sources. The amount of citrate synthase activity and immune-reactive protein increased more than 15-fold as S. cerevisiae cells entered the stationary growth phase on glucose-containing medium. This increase was paralleled by an increase in translatable RNA for the enzyme. When cells were grown on a nonfermentable carbon source, no increase in either citrate synthase or its mRNA was detected. The results suggest that the release of citrate synthase from catabolite repression may occur at the level of transcription.

Citrate (si)-Synthase↗
PubMed1983

Investigation of white-rot fungi for the conversion of poplar into a potential feedstuff for ruminants.

Five isolates of wood-rotting fungi were compared for their ability to increase the digestibility of poplar shavings. Homogenates of the fungi were inoculated into mixtures of 25 g of shavings and 100 mL of supplementary nutrient solution and incubated at 25 degrees C. The in vitro rumen digestibility of the products increased from 30% at the start of the fermentation to maxima of 72% with Polyporus anceps after 8 weeks, 64% with Ganoderma applanatum and 62% with Phanerochaete chrysosporium after 4 weeks, 61% with Polyporus versicolor after 3 weeks, and 42% with Fomitopsis ulmarius after 4 weeks. Fibre analysis showed a decrease in lignin as digestibility increased. Loss of carbohydrates occurred in all fermentations and continued after lignin loss ceased. Dry matter yield at the time of maximum digestibility ranged from 80 to 94% of the original dry matter. The highest digestibilities attained were similar to those of conventional roughage feeds for ruminants. This shows that this fermentation could form the basis of a practical system for converting poplar into a feedstuff.

Animal Feed↗
PubMed1980

Protective effect of milk against in vitro caries.

Additions of milk solids to laboratory "cakes" made of sucrose and starch reduced in vitro (Orofax) "caries" and the dissolution of granular enamel, but increased the amounts of fermentation acids produced. Withdrawal of the milk component of ice cream, yogurt, and chocolate milk resulted in increased Orofax "caries".

Acids↗
PubMed1997

[Discovery and development of a novel immunosuppressant, tacrolimus hydrate].

Tacrolimus hydrate (FK506), a novel 23-membered macrolide, is an immunosuppressant isolated from Streptomyces tsukubaensis using extensive screening of fermentation products to identify a compound inhibiting the mixed lymphocyte reaction (MLR). The in vitro and in vivo immunosuppressive activities of FK506 were found to be more potent than those of cyclosporine (CyA). The superior immunosuppression with FK506 treatment was also confirmed in the skin allograft model in rats and liver transplantation in dogs. Clinical studies were initiated by Prof. Starzl at the University of Pittsburgh in 1989, and he demonstrated that FK506 surpassed CyA in the incidence of graft survival and the frequency of graft rejection. Multicenter randomized clinical studies, comparing FK506 to CyA corroborated the efficacy of FK506 on the survival of patients and of grafts, and especially on the appearance of severe refractory graft rejection. FK506 was marketed in 1993 in Japan, and was followed in 1994 in the U.S.A., U.K. and Germany. The mechanism of action of this compound was clarified by the endeavors of Prof. Schreiber, who demonstrated the existence of a binding protein for FK506 called FKBP, similar to cyclophilin for CyA. The FK506/FKBP complex binds with calcineurin, a serine/threonine phosphatase to inhibit the translocation of NFAT into the nucleus, resulting in inhibition of transcription of IL-2 mRNA. FK506 displays potent immunosuppressant activity, and contributes not only to the progress of transplantation therapy for clinical studies, but also to the clarification of signal transduction in T cell activation for basic science.

Animals↗
PubMed2003

Biohydrogenation of unsaturated fatty acids in continuous culture fermenters during digestion of orchardgrass or red clover with three levels of ground corn supplementation.

Diet digestibility and outputs of biohydrogenation intermediates were assessed in a continuous culture of ruminal microorganisms. Orchardgrass or red clover harvested and frozen during spring or fall served as the primary substrates for fermentation. During 10-d incubations, fermenters were fed thawed forage (50 g of DM/d), forage (42 g/d) plus 8 g/d of corn, or forage (34 g/d) plus 16 g/d of corn. Effluents from the last 3 d of incubation were composited for analyses. Starch input increased from 5 to 27% of DM as corn input increased from 0 to 16 g/d. Corn input reduced (P < 0.01) pH, increased (P < 0.01) microbial DM yield, and increased (P = 0.01) digestibility of DM, NDF, CP, and nonstructural carbohydrates. Overall, apparent hydrogenation (percentage) of cis9-18:1, 18:2n-6, and 18:3n-3 was greater (P < 0.05) with orchardgrass than clover. Hydrogenation of cis9-18:1 and 18:2n-6 increased (P = 0.01), but hydrogenation of 18:3n-3 decreased (P = 0.01) linearly due to corn input, regardless of forage. As a result, output of trans11, cis15-18:2 also decreased (P = 0.01). Average output of cis9,trans11-18:2 was greater (P = 0.01) for clover (1.3 mg/d) compared with orchardgrass (0.6 mg/d), but corn input with either forage increased (P = 0.01) cis9,trans11-18:2 output by 205%. Output of trans11-18:1 was greater (P = 0.01) from orchardgrass compared with clover (174 vs. 90 mg/d), but corn increased (P = 0.01) trans11-18:1 output only from clover fermentations. Output of trans10-18:1 was greater (P = 0.01) in response to orchardgrass compared with clover (10 vs. 4 mg/d), but corn addition doubled the output regardless of forage type. Output of trans10,cis12-18:2, which did not differ due to forage type, increased (P = 0.01) twofold in response to corn. Cis9,cis11-18:2 was a primary conjugated isomer produced from forage fermentations, but its output decreased (P = 0.03) in response to corn input. When inputs of 18:2n-6 plus 18:3n-3 were less than 0.9% of total DM (clover), hydrogenation was low (87%). When 18:2n-6 plus 18:3n-3 inputs were from 1.2 to 1.5% of total DM (orchardgrass), hydrogenation averaged 96%. Despite greater hydrogenation, incremental additions of cis9-18:1 and 18:2n-6 from corn grain increased (P < 0.05) outputs of trans10-18:1, trans11-18:1, trans10,cis12-18:2, cis9,trans11-18:2, and trans,trans-18:2 in effluent. Results suggest that forage species alone or in combination with corn grain can alter hydrogenation and profiles of intermediates to varying degrees.

Animal Feed↗
PubMed1986

Grain processing effects on starch utilization by ruminants.

Starch utilization may be markedly enhanced by proper grain processing; however, extent of improvement is primarily dependent upon the ruminant species, grain source and method of processing. Grain processing has less impact on starch digestion by sheep than cattle. The magnitude of improvement is inverse to the starch digestion values for nonprocessed (or minimally processed) grains. Utilization of sorghum grain starch is improved most by extensive processing, and then corn, with little improvement in barley starch digestion. Studies comparing processing effects on barley or wheat starch utilization by cattle were not found. Steam-flaking consistently improves digestibility of starch by cattle fed corn- or sorghum grain-based diets over whole, ground or dry-rolled processes. Other extensive processing methods appear to enhance starch digestibility of corn and sorghum grain to a similar extent as steam-flaking, but comparative data are too limited to quantitate adequately effects of these methods. This improvement in starch utilization appears to be the primary reason for enhanced feed conversion of cattle fed diets high in these processed grains. The major site of cereal grain starch digestion is usually the rumen. Processing increases microbial degradation of starch in the rumen and decreases amounts of starch digested post-ruminally. Rates of in vitro amylolytic attack of starch in cereal grains by both ruminal microbial and pancreatic enzyme sources are improved by processing methods employing proper combinations of moisture, heat and pressure. In vitro and in situ studies suggest that much of the increase in ruminal starch fermentation with steam-flaking is due to changes in starch granular structure, which produces additive effects beyond those of decreasing particle size. Thus, efficiency of ruminal starch fermentation by cattle appears to be improved by proper processing of corn and sorghum grain. Processing and grain source studies both suggest that maximal total tract starch digestibility is positively related to the extent of digestion in the rumen.

Animal Feed↗
PubMed1995

Inability of Lactobacillus casei strain GG, L. acidophilus, and Bifidobacterium bifidum to degrade intestinal mucus glycoproteins.

BACKGROUND: Lactic acid bacteria have been suggested for use in the prevention of relapse of ulcerative colitis and of recurrent pouchitis. These strains may not damage the protective intestinal mucus glycoproteins. METHODS: Lactobacillus casei strain GG and strains isolated from a commercial fermented product (Lactobacillus acidophilus, Bifidobacterium bifidum, and a mesophylic lactic culture) were cultured in vitro on hog gastric mucin and human intestinal glycoproteins. Furthermore, germ-free rats were mono-associated with Lactobacillus GG and poly-associated with the other strains. Glycoproteins were isolated from rat distal ileum, cecum, and colon. Mucus degradation was established by assaying carbohydrates (hexosamines, hexoses, pentoses), proteins, and blood group antigenicity. RESULTS: All strains colonized the intestinal mucus but were not found in the deep crypts. Degradation of mucus glycoproteins was observed neither in vitro nor in vivo. CONCLUSION: The tested strains do not break down intestinal mucus glycoproteins and thus far are safe to use for therapy.

Animals↗
PubMed1977

Mutational biosynthesis by idiotrophs of Micromonospora purpurea. II. Conversion of non-amino containing cyclitols to aminoglycoside antibiotics.

A mutant of Micromonospora purpurea, which produces the gentamicin complex only when 2-deoxystreptamine is added to the fermentation medium, produces a new antibiotic complex, 2-hydroxygentamicin, when streptamine or 2,4,6/3,5-pentahydroxycyclohexanone is added to the fermentation medium. This mutant also produces the gentamicin complex when 2,4/3,5-tetrahydroxycyclohexanone is added to the fermentation medium. The C1 and C2 components of 2-hydroxygentamicin have broad spectrum in vitro antibacterial activity similar to the gentamicin C1 and C2 components, but with greater activity against some gentamicin-resistant strains.

Bacteria↗
PubMed1995

Simple aromatics identified with a NFAT-lacZ transcription assay for the detection of immunosuppressants.

Determination of the mechanism of action of FK506 and cyclosporin A has yielded new molecular targets involved in signal transduction during T cell activation. A common target of FK506 and cyclosporin A is inhibition of activation of the NFAT transcription factor, for which a specific binding region is present in the promoter of the IL-2 gene. A reporter gene assay has been used to screen for agents that interfere with this early step in T cell activation. Simple aromatic compounds that block NFAT-dependent transcription and show in vitro immunosuppressive activity were isolated from the broth and mycelia of two Streptomyces sp. fermentations. The compounds were active at concentrations that were not directly cytotoxic.

Animals↗