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Biomedical subjects

O Simon

Publications and source records attributed to O Simon.

At least 37 records · Page 2Linked to original sources

The measurement of muscle protein synthesis in broilers with a flooding dose technique: use of 15N-labelled phenylalanine, GC-MS and GC-C-IRMS.

An experiment was carried out to measure fractional muscle protein synthesis rates (k(s)) in broilers with injection of a flooding dose of phenylalanine (1 ml/100 g body weight of 150 mM phenylalanine; 38 atom percent excess (APE) [15N]phenylalanine). K(s) was calculated from the [15N] enrichment in phenylalanine of tissue-free and protein-bound phenylalanine using both gas chromatography mass spectrometry (GC-MS) and gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) for measurements after a 10 min isotope incorporation period. The tertiary-butyldimethylsilyl (t-BDMS) derivatives of phenylalanine were used for gas chromatographic separation in both systems. GC-MS and GC-C-IRMS were calibrated for a range of 7 to 37 [15N]APE and 0 to 0.62 [15N]APE, respectively, and for sample sizes of 0.45 to 4.5 nmol phenylalanine and 7 to 40 nmol phenylalanine, respectively. Reproducibility of standards as a measure of precision varied from 0.06 to 0.29 [15N]APE and from 0.0004 to 0.0018 [15N]APE in GC-MS and GC-C-IRMS, respectively. K(s) was measured in the m. pectoralis major of broilers fed rye based diets (56%) which were provided either unsupplemented (-) or supplemented (+) with an enzyme preparation containing xylanase. K(s) in breast muscles was significantly increased from 21.8%/d to 23.9%/d due to enzyme supplementation. It can be concluded from the study that the measurement of protein synthesis in broilers with the flooding dose technique can be carried out by using [15N]phenylalanine, GC-MS and GC-C-IRMS.

Animals↗

Growth behaviour of a spore forming probiotic strain in the gastrointestinal tract of broiler chicken and piglets.

The growth behaviour of the probiotic strain Bacillus cereus var. toyoi in the gastrointestinal tract of broiler chicken and suckling piglets was evaluated. The strain germinated rapidly in intestinal samples from both animal species. Less than 10% of spores were recovered from the chicken crop and piglet stomach, respectively. Lumen samples and mucosal tissues from the hind gut of piglets displayed increasing colonization of the probiotic strain throughout the trial period. After oral administration of vegetative cells to broiler chicken and weaned piglets, sporulation was detected in all intestinal samples. The distribution of spore CFU indicated repeated germination and sporulation during the intestinal passage in piglets. B. cereus var. toyoi was not able to colonize the intestinal tract of both animal species. However, the probiotic strain was detected in suckling piglets before uptake of B. cereus var. toyoi supplemented feed. It is concluded that B. cereus var. toyoi germinates rapidly in broiler chicken and piglets, which is a necessary prerequisite for its possible probiotic effects. Germination and in vivo sporulation of vegetative cells indicated that the probiotic strain was metabolically active in the intestine of both animal species.

Administration, Oral↗

The effectiveness of an Escherichia coli phytase in improving phosphorus and calcium bioavailabilities in poultry and young pigs.

The effectiveness of an Escherichia coli phytase in comparison with a commercially available Aspergillus phytase in improving the bioavailability of phosphorus in broilers, layers and young pigs was studied in three separate experiments. Three basal diets, marginally deficient in dietary P mainly provided as phytate, were formulated. Both phytases were added to the diets at the rate of 500 U/kg diet. The phytases significantly (P < or = 0.05) improved the availability of phytate P to broilers, layers and young pigs. Aspergillus and E. coli phytases enhanced the pre-caecal digestibility of P by 11 and 29% for broilers and 18 and 25% for layers, respectively. Total tract digestibility of P (P balance) was also enhanced but with smaller magnitude. In pigs, total tract digestibility of P was improved by 33 and 34% by Aspergillus and E. coli phytases, respectively. Under the conditions of this study, it was observed that E. coli consistently, though with small magnitude in layers and pigs, enhanced the availability of phytate P at the same range or slightly better than Aspergillus phytase. It was only in pigs that the availability of Ca was significantly (P < or = 0.05) improved by addition of both phytases. It can be concluded that E. coli phytase is highly effective in improving the bioavailability of phytate P to broilers, layers and young pigs. This seems to be based on the high proteolytic stability of the enzyme in the digestive tract, as shown recently.

6-Phytase↗

Simultaneous experimental study of direct and direct plus trophic contamination of the crayfish Astacus astacus by inorganic mercury and methylmercury.

An experimental study was carried out to investigate direct and direct plus trophic contamination routes of the crayfish Astacus astacus by inorganic mercury (Hg(II)) or methylmercury (MeHg). Direct exposure was based on low contamination conditions, 300 and 30 ng/L in the dissolved phase, respectively, during 30 d at 20 degrees C. Trophic exposure was based on daily consumption of the Asiatic clam Corbicula fluminea, previously contaminated during 40 d with similar exposure conditions. The Hg concentrations in the bivalves were very similar: 1,451 +/- 287 ng/g for Hg(II) and 1,346 +/- 143 ng/g for MeHg. In the crustaceans, Hg bioaccumulation was analyzed at the whole-organism level and in eight organs (gills, stomach, intestine, hepatopancreas, tail muscle, green gland, carapace, and hemolymph), after 15 and 30 d of exposure. Analysis of the results showed marked differences between Hg(II) and MeHg accumulation in favor of MeHg: for the direct route, the ratio between metal concentrations was close to 8; for the trophic route, no significant increase in Hg accumulation was observed for Hg(II) even when the ratio between Hg concentration in the direct plus trophic contamination route and Hg concentration in the direct contamination route was 1.6 for MeHg, with an estimated trophic transfer rate close to 20%. Mercury organotropism was also specifically connected to the exposure conditions, especially at the biological barrier level according to the route of exposure: gills and carapace for the direct route and digestive tract including hepatopancreas for the trophic route.

Animals↗

Isolation of an 1,3-1,4-beta-glucan degrading Enterococcus faecium strain from the intestinal tract of chicken and partial characterization of its novel 1,3-1,4-beta-glucanase.

An Enterococcus faecium strain with a novel endo 1,3-1,4-endo-beta-glucanase (lichenase, E.C. 3.2.1.73) was isolated from the intestinal tract of broiler chicken. The enzyme was secreted into the culture medium and acted exclusively on mixed linked 1,3-1,4-beta-glucans as determined with a reducing sugar assay. The purified enzyme has its isoelectric point at pI 4.8, maximum activity was determined at pH 6.5 and 40 degrees C. Thermal stability of the enzyme was low, but high pH stability and high residual activity was observed after incubation in digesta samples from the chicken intestine. Multiple lichenase activities were obtained from culture supernatants on SDS/PAGE and native zymograms, but it is concluded that the lichenase consists of one active protein at 30.5 kD and additional polypeptides of unknown function.

Animals↗

Comparative experimental study of cadmium and methylmercury trophic transfers between the asiatic clam Corbicula fluminea and the crayfish Astacus astacus.

Cadmium (Cd) and methylmercury (MeHg) trophic transfers were analyzed between the Asiatic clam Corbicula fluminea and the crayfish Astacus astacus. Metal bioaccumulation in crayfish was quantified after 5, 10, and 15 days of exposure via daily ingestion of soft bodies of C. fluminea, previously exposed during 7 days to Cd (20 microg. L(-1)) and MeHg (4 microg. L(-1)). Bioaccumulation kinetics in the predator were investigated at organ and tissue levels: hemolymph, tail muscle, hepatopancreas, gills, stomach/mesenteron, intestine, green gland, carapace. Trophic transfer rates were estimated at the whole organism level. Results showed marked differences (1) in assimilation efficiencies, mean transfer rates being 5% for Cd and 16% for MeHg; and (2) in the metal distribution within the different tissue compartments of the crayfish: for Cd, the trophic uptake leads to high concentrations in the hepatopancreas and small accumulation in the muscle tissue; for MeHg, the highest levels of bioaccumulation occur in the green gland and in the tail muscle. From an ecotoxicological point of view, these experimental data suggest a small risk of Cd transfer between the crayfish and predators, humans included; on the other hand, Hg distribution in the muscle and accumulation trends in this tissue represent an obvious risk of transfer.

Animals↗

Endogenous N-losses in broilers estimated by a [15N]-isotope dilution technique: effect of dietary fat type and xylanase addition.

Male broilers were given a low protein diet (15.5% CP) spiked with [15N]H4HCO3 from day 12 to day 18 of age to label the endogenous N-constituents. Experimental diets were subsequently fed from day 19 to day 24 of age and consisted of a rye based diet (56% dietary inclusion) which contained either 10% soya oil (S) or 10% beef tallow (T), each of which was either unsupplemented (-) or supplemented (+) with a xylanase containing enzyme preparation (2700 IU/kg at pH 5.3). [15N]-atom percent excess (APE) of excreta, faeces and urine were monitored on a daily basis during both experimental periods. Furthermore, APE was measured in various tissues at the end of the experiment. The APE of urine on the last day of the experiment was between the APE of the pancreas and that of the jejunal tissue, an observation which supported the usefulness of using urinary APE as an indicator for the endogenous N-pool. Endogenous N-proportions were estimated by an isotope dilution technique at the end of the experiment by examination of the ratio of APE in faeces and urine. The endogenous N-proportion in the faeces was greatest in birds receiving the T(-) diet. The proportions were 0.321, 0.319, 0.451 and 0.289 in S(-), S(+), T(-) and T(+) fed groups, respectively. Xylanase addition reduced endogenous N-proportion, a factor which was used to correct apparent crude protein digestibility (85.6, 86.2, 84.3 and 88.5% in S(-), S(+), T(-) and T(+) fed birds, respectively) for endogenous losses resulting in almost equal true digestibilities of crude protein for all treatments (90.3, 90.6, 90.4 and 91.5%). The amounts of endogenous N in faces were estimated to be 87, 69, 244 and 81 mg per day per kg0.67 body weight in S(-), S(+), T(-) and T(+) fed birds, respectively. It was concluded that xylanase supplementation of a rye based broiler diet does not change endogenous N-secretions when the supplemental fat is soya oil. However, addition of tallow rather than soya oil increased these N-losses significantly, an effect which was reversed by xylanase addition.

Animal Feed↗

Comparative studies on the in vitro properties of phytases from various microbial origins.

The physical and chemical properties of six crude phytase preparations were compared. Four of these enzymes (Aspergillus A, Aspergillus R, Peniophora and Aspergillus T) were produced at commercial scale for the use as feed additives while the other two (E. coli and Bacillus) were produced at laboratory scale. The encoding genes of the enzymes were from different microbial origins (4 of fungal origin and 2 of bacterial origin, i.e., E. coli and Bacillus phytases). One of the fungal phytases (Aspergillus R) was expressed in transgenic rape. The enzymes were studied for their pH behaviour, temperature optimum and stability and resistance to protease inactivation. The phytases were found to exhibit different properties depending on source of the phytase gene and the production organism. The pH profiles of the enzymes showed that the fungal phytases had their pH optima ranging from 4.5 to 5.5. The bacterial E. coli phytase had also its pH optimum in the acidic range at pH 4.5 while the pH optimum for the Bacillus enzyme was identified at pH 7.0. Temperature optima were at 50 and 60 degrees C for the fungal and bacterial phytases, respectively. The Bacillus phytase was more thermostable in aqueous solutions than all other enzymes. In pelleting experiments performed at 60, 70 and 80 degrees C in the conditioner, Aspergillus A, Peniophora (measurement at pH 5.5) and E. coli phytases were more heat stable compared to other enzymes (Bacillus enzyme was not included). At a temperature of 70 degrees C in the conditioner, these enzymes maintained a residual activity of approximately 70% after pelleting compared to approximately 30% determined for the other enzymes. Incubation of enzyme preparations with porcine proteases revealed that only E. coli phytase was insensitive against pepsin and pancreatin. Incubation of the enzymes in digesta supernatants from various segments of the digestive tract of hens revealed that digesta from stomach inactivated the enzymes most efficiently except E. coli phytase which had a residual activity of 93% after 60 min incubation at 40 degrees C. It can be concluded that phytases of various microbial origins behave differently with respect to their in vitro properties which could be of importance for future developments of phytase preparations. Especially bacterial phytases contain properties like high temperature stability (Bacillus phytase) and high proteolytic stability (E. coli phytase) which make them favourable for future applications as feed additives.

6-Phytase↗

Intra- and inter-individual variability in human sperm concentration, motility and vitality assessment during a workshop involving ten laboratories.

The aim of the present study was to assess variability in the evaluation of human sperm concentration, motility and vitality. Technicians and biologists from 10 teams involved in multicentre studies on semen quality attended the same laboratory, each team using its own methods and equipment to analyse the same semen samples. Inter-individual variability was assessed from 17 fresh semen samples of varying quality. Intra-individual variability was assessed from pools of frozen samples for sperm concentration and motility and stained smears for vitality with three blind evaluations by sample and smear. The mean inter-individual coefficients of variation were 22.9, 21.8 and 17.5% for sperm concentration, motility and vitality respectively. There was no statistical difference among participants for sperm concentration assessment, but significant differences for both motility and vitality (both P: < 0.05). The mean intra-individual coefficients of variation were 15.8, 26.2 and 13.1% for sperm concentration, motility and vitality respectively, with marked differences between expert and novice participants: concentration 9.8% versus 28.0%; motility 22.8% versus 33.0%; and vitality 10.0% versus 19.3%. The present data confirm the need for external quality control schemes for diagnostic purposes, and indicate their utmost importance in multicentre studies on semen quality.

Cell Survival↗

Replacement of soybean oil with tallow in rye-based diets without xylanase increases protein synthesis in small intestine of broilers.

We examined the effects of dietary fat type (10% of either soybean oil, S, or beef tallow, T)(3) and xylanase supplementation (-, without; +, with 1 g of Avizyme 1300 per kg diet) in rye- based diets (56%) on tissue protein synthesis in male broilers. Birds were injected with a large flooding dose of a phenylalanine solution (150 mmol/L, 38 atom percentage excess [(15) N] phenylalanine) and tissues were obtained after a 10-min incorporation period. [(15) N]-enrichment in tissue free phenylalanine and tissue protein bound phenylalanine were measured by gas-chromatography mass-spectrometry and by gas-chromatography combustion isotope ratio mass-spectrometry, respectively in order to calculate tissue specific fractional rates of protein synthesis (k(s)). The k(s) (%/d) in (S-), (S+), (T-) and (T+)-fed birds were 56, 64, 84 and 61 (SEM = 3.7) in duodenum, 51, 52, 75 and 58 in jejunum (SEM = 3.1), 66, 67, 105 and 68 (SEM =7.0) in jejunal mucosa cells, 53, 56, 68 and 50 (SEM = 3.7) in ileum and 52, 45, 118 and 39 (SEM = 20.2) in pancreas, respectively. Significant fat, enzyme or interaction effects in these tissues were mainly caused by the elevated k(s) in (T-)-fed birds which was closely associated with intestinal viscosity. We conclude that the effect of soluble nonstarch polysaccharides (NSP) and of NSP-hydrolyzing enzymes may be explained partially by modification in tissue protein synthesis of the intestinal tract.

Animals↗

Biochemical characteristics of non starch polysaccharide hydrolyzing enzyme preparations designed as feed additives for poultry and piglet nutrition.

Non starch polysaccharide hydrolyzing enzyme preparations analyzed in this study were composed of up to nine (1-3, 1-4)-beta-glucanase activities and up to six xylanase activities with different molecular weights in the range from 100 kD down to 18 kD as determined with SDS/PAGE zymograms. Partially purified enzyme fractions differed in terms of pH-optima, isoelectric point and thermal stability in aquaeous solutions. Different beta-glucanase activities were found in different production strains, although some enzymes were conserved over genus boundaries. Enzyme preparations from the same or related strains exhibited different patterns of enzyme activity, indicating modification of strain and/or fermentation conditions. Some enzyme preparations contained significant amounts of polygalacturonase and/or galactomannase activity. The pH profiles of whole enzyme preparations resulted from pH optima of isoenzyme fractions. Temperature optima for all preparations were between 50 and 60 degrees C. Thermal stability of high molecular weight components tended to be lower than for low molecular weight fractions. Fractions with cellulase activity were most stable, followed by (1-3, 1-4)-beta-glucanase activities, while fractions with xylanase activities exhibited low thermal stabilities. Incubation of enzyme preparations and their respective active fractions in digesta supernatants revealed only small differences in residual xylanase activity. Digesta from gizzard samples led to the highest inactivation. It is concluded that commercial enzyme preparations display different modes of action and that the development of improved enzyme preparations depends not only on thermal stability, but also on pH profile, substrate specificity and proteolytic stability within the digestive tract.

Animal Nutritional Physiological Phenomena↗

Effects of dietary fat type, pentosan level and xylanase supplementation on digestibility of nutrients and metabolizability of energy in male broilers.

A complete two by two by four factorial design was used to examine the main effects of dietary fat type (10% soya oil or 10% beef tallow), xylanase supplementation (with or without Avizyme 1300 at 1 g/kg diet) and pentosan level (7.7 g/kg, 11.0 g/kg, 14.3 g/kg and 17.6 g/kg soluble pentosans, respectively, by varying wheat/rye proportions) as well as their interactions on intestinal chyme conditions, nutrient digestibility and nutrient utilization in male broilers. Nutrient digestibilities for the total digestive tract and at various sites of small intestine were measured during the period from day 18 to 20 of age and at day 21 of age, respectively, using a marker technique. Jejunal and ileal supernatant viscosity increased in an exponential manner as dietary pentosan concentration was increased. This increase was more pronounced in tallow fed birds but was also found in enzyme treated groups albeit at a much lower level. Xylanase activity was still detectable in the ileum of birds fed enzyme supplemented diets but its activity was found to decrease as dietary pentosan content increased. Digestibility of crude protein and that of some amino acids at the terminal ileum was decreased as dietary pentosan content was increased and significantly improved by xylanase addition. No fat effect and no interactions were detected at this site. In contrast, measurements made over the whole gastrointestinal tract showed significantly lower protein and amino acid digestibility values for tallow fed birds, and significant higher enzyme effects especially in diets with higher pentosan concentrations. Nitrogen-corrected apparent metabolizable energy (AMEN) content and net protein utilization decreased with increasing dietary pentosan content and were significantly improved by xylanase addition and were lower in tallow fed birds. Again, xylanase effects were found to be more pronounced for tallow fed birds and at higher pentosan concentrations.

Amino Acids↗

Effects of dietary fat type and xylanase supplementation to rye-based broiler diets on selected bacterial groups adhering to the intestinal epithelium. on transit time of feed, and on nutrient digestibility.

Two experiments were conducted to examine the effects of different fat types, i.e., soybean oil (S) and beef tallow (T), in rye-based broiler diets, either unsupplemented (-) or supplemented (+) with xylanase (Avizyme 1300 at 1 g/kg diet), on selected bacterial groups adhering to the epithelium of crop, duodenum, jejunum, and ileum (Experiment 1, 16 d of age), on mean retention time (MRT) of digesta, and on digestibility of N and dry matter in successive segments of the digestive tract (Experiment 2, 24 d of age). Live weight of enzyme-treated and S-fed chickens was significantly higher than that for unsupplemented or T-fed birds, respectively, in both experiments. In Experiment 1, a reduction in bacterial colonization from crop to duodenum was followed by a continuous increase as far as the ileum. Xylanase supplementation significantly reduced enterobacteria and total anaerobe microbes with a similar trend for Gram-positive cocci and enterococci. The latter two groups were significantly enhanced in birds fed T. In Experiment 2, xylanase supplementation resulted in a decrease in MRT in several segments of the digestive tract. This effect was most pronounced in the small intestine, where MRT of 268, 217, 241, and 209 min in groups S-, S+, T-, and T+, respectively, were measured. Apparent digestibility of N and dry matter was slightly improved by xylanase supplementation in the jejunum and ileum. Nitrogen digestibility by the terminal ileum was 80.3, 83.7, 79.4, and 82.2% for the S-, S+, T-, and T+ groups, respectively, and dry matter digestibility amounted to 61.2, 65.5, 62.1, and 64.0%, respectively.

Animal Feed↗

Interactions between dietary fat type and xylanase supplementation when rye-based diets are fed to broiler chickens. 1. Physico-chemical chyme features.

1. The interactions between fat type and xylanase supplementation of rye-based diets were investigated using a 2 x 2 factorial design in which a rye-based diet (610 g rye/kg) was combined with 100 g/kg of soya oil or beef tallow, with and without xylanase supplementation at 3000 IU/kg, and fed to 1-d-old male broilers. Food passage time, viscosity of digesta supernatant, xylanase activity and pH in different segments of the digestive tract were examined. 2. Food passage throughout the digestive tract was accelerated by enzyme addition regardless of fat type. The time taken for 50% of the marker to be excreted was reduced from 8.4 to 6.7 h in animals receiving the rye-soya oil diets and from 8.0 to 6.9 h with the rye-tallow diets. 3. Viscosity in the supernatant of the jejunal and ileal digesta was markedly decreased after enzyme addition. Viscosities were generally higher in the ileal than in the jejunal supernatant, and fell as the birds aged from 14 to 28 d. The effect of enzyme was also reduced in older chicks. There was not a clear effect of the fat source on viscosity. 4. Xylanase activity was still found at the end of the ileum in digesta of birds fed on the enzyme-supplemented diets but not in control animals. Xylanase activity was also detected in the caeca of all groups. 5. Significantly lower pH values were found in tallow-fed birds in some segments of the digestive tract. A significant increase in pH after enzyme addition was detected in the proximal ileum; this was independent of fat source.

Animal Feed↗

Interactions between dietary fat type and xylanase supplementation when rye-based diets are fed to broiler chickens 2. Performance, nutrient digestibility and the fat-soluble vitamin status of livers.

1. The interactions between dietary fat type and xylanase supplementation of rye-based diets were investigated using a 2 x 2 factorial design in which a rye-based diet (610 g rye/kg) was combined with 100 g/kg of soya oil or beef tallow, with or without xylanase supplementation at 3000 IU/kg, and fed to 1-d-old male broilers for 35 d. Growth, nutrient digestibility and AMEN values were determined and the vitamin status of livers assessed at various ages. 2. There were significant interactions between crude fat and xylanase supplementation on the faecal digestibility of crude fat and crude protein. Fat digestibility was improved by xylanase in both fat type groups but to a greater extent for the tallow diets. However, the effects were found to be significant only for oleic, linoleic and linolenic acids. Similarly, protein digestibility and AMEN values were significantly improved by xylanase, but only for the tallow diet. 3. Ileal digestibility of nitrogen and amino acids also were affected by enzyme for both fat type diets, the effect generally being more pronounced for the tallow diet. 4. The deposition of the fat soluble vitamins A and E in livers was significantly increased by xylanase supplementation and was also better for soya oil than for tallow. 5. Xylanase supplementation increased the digestibility of insoluble pentosans whereas for the soluble pentosans the opposite effect was noted. 6. Without xylanase supplementation the tallow-based diet caused high mortality. Liveweight gain and the efficiency of food utilisation were greatest in the soya oil-based, xylanase-supplemented diet, followed by the unsupplemented soya oil-, supplemented tallow- and unsupplemented tallow-based diets.

Amino Acids↗

An investigation into the variability of extract viscosity of wheat-relationship with the content of non-starch-polysaccharide fractions and metabolisable energy for broiler chickens.

The in vitro extract-viscosity and the content of non-starch-polysaccharides were investigated in 34 defined wheat varieties grown at 5 locations each. Both, wheat genotype as well as growing location clearly influenced the viscosity of soluble extract from wheat. Furthermore, the content of non-starch-polysaccharides (soluble/total) and pentosans (soluble/total) were determined in 13 wheat varieties each grown at two locations. Soluble pentosan contents were highly positively correlated with extract viscosity of wheat at the locations Hayn (r = 0.86) and Biendorf (r = 0.90). The classical apparent metabolisable energy of 5 wheat samples having different extract viscosities was assessed. The AMEN values ranged from 14.0 to 14.6 MJ/kg DM and were significant negatively correlated to content of soluble arabinoxylans (r = 0.67) and to the extract viscosity (r = 0.83). Furthermore, the viscosity of jejunal (4.0 to 22.8 mPas) and ileal (13.1 to 78.0 mPas) digesta exhibited a clear relationship with soluble pentosan contents and extract viscosity. Under the conditions applied in this study the technique of extract viscosity measurement can predict the AME.

Analysis of Variance↗