Metabolic Effects of a Hydrophobic Alginate Derivative and Tetrahydrolipstatin in Rats Fed a Diet Supplemented with Palm Fat and Cholesterol.

The effects of octadecylamide of alginic acid (amidated alginate) and tetrahydrolipstatin on serum and hepatic cholesterol, and the faecal output of fat and sterols, were investigated in rats. Amidated alginate is a sorbent of lipids, tetrahydrolipstatin is an inhibitor of pancreatic lipase. Rats were fed diets containing cholesterol and palm fat at 10 and 70 g/kg, respectively. Palm fat was provided by coconut meal. Amidated alginate at 40 g/kg diet significantly decreased serum total cholesterol, low-density lipoprotein and hepatic cholesterol, and hepatic lipids and increased the faecal output of fat and coprostanol. Tetrahydrolipstatin at 300 mg/kg diet significantly decreased low-density lipoprotein cholesterol and hepatic lipids and increased the faecal output of fat. The intake of feed was not significantly influenced; however, the weight gains in rats fed amidated alginate were lower than in rats of the control group. Both amidated alginate and tetrahydrolipstatin modified the fatty acid profile in excreta lipids. Concentrations of saturated fatty acids were decreased and those of unsaturated fatty acids increased. Despite different modes of action, amidated alginate and tetrahydrolipstatin were equally efficient in removing the dietary fat from the body.

Effects of pure plant secondary metabolites on methane production, rumen fermentation and rumen bacteria populations in vitro.

In this study, the effects of seven pure plant secondary metabolites (PSMs) on rumen fermentation, methane (CH4 ) production and rumen bacterial community composition were determined. Two in vitro trials were conducted. In trial 1, nine concentrations of 8-hydroxyquinoline, α-terpineol, camphor, bornyl acetate, α-pinene, thymoquinone and thymol were incubated on separate days using in vitro 24-hr batch incubations. All compounds tested demonstrated the ability to alter rumen fermentation parameters and decrease CH4 production. However, effective concentrations differed among individual PSMs. The lowest concentrations that reduced (p < .05) CH4 production were as follows: 8 mg/L of 8-hydroxyquinoline, 120 mg/L of thymoquinone, 240 mg/L of thymol and 480 mg/L of α-terpineol, camphor, bornyl acetate and α-pinene. These concentrations were selected for use in trial 2. In trial 2, PSMs were incubated in one run. Methane was decreased (p < .05) by all PSMs at selected concentrations. However, only 8-hydroxyquinoline, bornyl acetate and thymoquinone decreased (p < .05) CH4 relative to volatile fatty acids (VFAs). Based on denaturing gradient gel electrophoresis analysis, different PSMs changed the composition of bacterial communities to different extents. As revealed by Ion Torrent sequencing, the effects of PSMs on relative abundance were most pronounced in the predominant families, especially in Lachnospiraceae, Succinivibrionaceae, Prevotellaceae, unclassified Clostridiales and Ruminococcaceae. The CH4 production was correlated negatively (-.72; p < .05) with relative abundance of Succinivibrionaceae and positively with relative abundance of Ruminococcaceae (.86; p < .05). In summary, this study identified three pure PSMs (8hydroxyquinoline, bornyl acetate and thymoquinone) with potentially promising effects on rumen CH4 production. The PSMs tested in this study demonstrated considerable impact on rumen bacterial communities even at the lowest concentrations that decreased CH4 production. The findings from this study may help to elucidate how PSMs affect rumen bacterial fermentation.

In vitro Screening of Essential Oil Active Compounds for Manipulation of Rumen Fermentation and Methane Mitigation.

The objective of this study was to investigate the effects of 11 active compounds of essential oils (ACEO) on rumen fermentation characteristics and methane production. Two trials were conducted. In trial 1, ACEO (eugenol, carvacrol, citral, limonene, 1,4-cineole, p-cymene, linalool, bornyl acetate, α-pinene, and ß-pinene) at a dose of 1,000 µL/L were incubated for 24 h in diluted rumen fluid with a 70:30 forage:concentrate substrate (16.2% crude protein; 36.6% neutral detergent fiber). Three fistulated Holstein cows were used as donors of rumen fluid. The reduction in methane production was observed with nine ACEO (up to 86% reduction) compared with the control (p<0.05). Among these, only limonene, 1,4-cineole, bornyl acetate, and α-pinene did not inhibit volatile fatty acid (VFA) production, and only bornyl acetate produced less methane per mol of VFA compared with the control (p<0.05). In a subsequent trial, the effects on rumen fermentation and methane production of two concentrations (500 and 2,000 µL/L) of bornyl acetate, the most promising ACEO from the first trial, were evaluated using the same in vitro incubation method that was used in the first trial. In trial 2, monensin was used as a positive control. Both doses of bornyl acetate decreased (p<0.05) methane production and did not inhibit VFA production. Positive effects of bornyl acetate on methane and VFA production were more pronounced than the effects of monensin. These results confirm the ability of bornyl acetate to decrease methane production, which may help to improve the efficiency of energy use in the rumen.

Effect of diets containing whole white lupin seeds on rabbit doe milk yield and milk fatty acid composition as well as the growth and health of their litters.

The effect of dietary inclusion of white lupin seed (WLS) on the milk composition and yield of rabbit does as well as the performance of their litters was studied. Two lactation diets having identical digestible protein (DP):DE ratio and two weaning diets having identical DP:DE ratio were formulated. The first lactation diet (SL) contained soybean meal (SBM; 13.0%) and sunflower meal (5.0%) as the main CP sources, whereas the second lactation diet (LL) was based on WLS (25.0%). As a result, the LL diet had a greater ether extract (EE) content than did the SL diet. The first weaning diet (SW) included SBM (7.0%) as the main CP source, whereas the second weaning diet (LW) diet was based on WLS (12.0%). No additional fat was added to any of the diets. A total of 32 (16 per treatment) Hyplus PS 19 does (4,225 ± 607 g BW, at the second parturition) were fed 1 of the 2 lactation diets. The litters were standardized to 9 kits (564 ± 81 g BW) on the day of birth and were fed 1 of the 2 weaning diets from d 17 to 69 of age. At d 30 of age (weaning), 66 rabbits on each weaning diet (689 ± 71 g BW; 3 per cage) were used to evaluate performance. Feed intake and doe BW were not affected by the dietary treatments. Milk yield tended to be higher between d 1 and 30 of lactation in does fed the LL diet (P = 0.094), a finding that is related to the higher dietary EE content and intake in the LL diet. When expressed per kilogram of metabolic weight, milk output (P < 0.05) and fat output (P < 0.05) were greater in these does. Improved G:F (P < 0.05) between d 1 and 21 of lactation and greater ADG (P = 0.072) and milk efficiency (P < 0.05) of litters was observed in does fed the LL diet. The milk of does fed the LL diet contained less linoleic acid (P < 0.05) and arachidonic acid (C 20:4n-6; P < 0.05) and more oleic acid (P < 0.05), α-linolenic acid (P < 0.05), and eicosapentaenic acid (P < 0.05), with a corresponding increase in the total PUFA n-3:C 20:4n-6 ratio (P < 0.05). The performance of fattening rabbits was not affected by dietary treatment. The number of ill plus dead rabbits caused by digestive disease was lower (P < 0.05) in rabbits fed the LW diet. Therefore, WLS is a suitable dietary CP source for lactating does that can replace traditionally used CP sources without adverse effects on feed intake and milk yield or on the growth and viability of their litters. Due to its fatty acid (FA) composition, the use of WLS in the lactation diet has the potential to improve the milk FA composition of does.

Surveillance and characterisation of Cronobacter spp. in Czech retail food and environmental samples.

Cronobacter spp. (formerly Enterobacter sakazakii) are emerging, opportunistic pathogens that are linked with food-borne infections in neonates and infants. In the present study, 291 samples of food, 36 samples from a dairy farm and 140 samples of dust from vacuum cleaners were examined for the presence of Cronobacter spp. using chromogenic media and biochemical tests. Altogether, 72 Cronobacter spp. strains were isolated in accordance with the reference standard CSN P ISO/TS 22964 (2006). No Cronobacter spp. strains were detected in 10 samples of infant milk formula or in samples from a dairy farm. Twelve out of 20 positive food samples were dry products. The incidence of Cronobacter spp. in instant and powdered products and spices (12 positive isolates out of 82 samples) was significantly higher than that in other foods (P = 0.002), but lower than that in samples of dust (52 isolates; P < 0.001). The incidence of Cronobacter spp. in dust from restaurants, bars and hotels (13 positive isolates in 20 samples) was significantly higher than that in dust from households (P = 0.010). The polymerase chain reaction assay for the species-specific detection of the rpoB gene was performed in 49 isolates. Thirty-four Cronobacter spp. isolates were identified as Cronobacter sakazakii, nine isolates as Cronobacter malonaticus and one isolate as Cronobacter turicensis.

Fermentation of mucin by bifidobacteria from rectal samples of humans and rectal and intestinal samples of animals.

Bifidobacteria (246 strains in total) were isolated from rectal samples of infants and adult humans and animals, and from intestinal samples of calves. Twenty-five strains grew well on mucin: 20 from infants, two from adults, and three from goatlings. Poor or no growth on mucin was observed in 156 bifidobacterial strains of animal origin. The difference between human and animal isolates in ability to grow on mucin was significant at p < 0.001. Nine human strains with the best growth on mucin were identified as Bifidobacterium bifidum. These strains produced extracellular, membrane-bound, and intracellular mucinases with activities of 0.11, 0.53, and 0.09 µmol/min of reducing sugars per milligram of protein, respectively. Membrane-bound mucinases were active between pH 5 and 10. The optimum pH of extracellular mucinases was 6-7. Fermentation patterns in cultures grown on mucin and glucose differed. On mucin, the acetate-to-lactate ratio was higher than in cultures grown on glucose (p = 0.012). We showed that the bifidobacteria belong to the mucin-fermenting bacteria in humans, but their significance in mucin degradation in animals seems to be limited.

Effect of coated and non-coated fatty acid supplementation on broiler chickens experimentally infected with Campylobacter jejuni.

The aim of this study was to examine whether and to what extent the supplementation of feed with a coated or non-coated mixture of fatty acids (caprylic and capric acid) affects broiler chickens experimentally infected with Campylobacter jejuni. The study was carried out using 48 chickens divided into four experimental groups. Throughout the whole rearing period (1-42 days), the chickens were fed a diet supplemented with 0.25% caprylic and capric acid (1:1), coated or non-coated. At the age of 14 and 28 days, chickens were orally challenged with C. jejuni. At regular time intervals post-inoculation, the shedding of C. jejuni was assayed using quantitative real-time PCR. Both supplements significantly decreased faecal C. jejuni counts by 1.2-4.1 log(10) CFU/g 4 days post-inoculation; after this time period, the effect of medium-chain fatty acids (MCFA) was less pronounced or absent. Campylobacter jejuni counts in excreta samples were significantly lower in chickens fed coated MCFA than in those fed non-coated MCFA. No effect of MCFA on feed intake or growth of chickens was observed. In conclusion, (i) MCFA are active against C. jejuni and (ii) the encapsulation enhanced the efficacy of the acids. These results allow the recommendation of using MCFA as feed additives in chickens, preferably 2-3 days before slaughter.

Susceptibility of Campylobacter jejuni to organic acids and monoacylglycerols.

Organic acids can be used as feed supplements or for treatment of poultry carcasses in processing plants. The antimicrobial activity of nineteen organic acids and two monoacylglycerols in cultures of Campylobacter jejuni CCM 6214(T) (ATCC 33560) was determined using a SYBR Green-based real-time PCR assay. The IC(50) was a concentration at which only 50 % of a bacteria specific DNA sequence was amplified. Caprylic, capric and lauric acids were the most efficient antimicrobials among the compounds tested (IC(50) < or = 0.1 mg/mL). In a weakly acidic environment (pH 5.5), the antimicrobial activity was more pronounced than at pH 6.5. At pH 5.5, oleic and fumaric acid also had clear antimicrobial activity, as did monocaprylin. The antimicrobial activity of acetic, butyric, stearic and succinic acid was low. In cells treated with fumaric acid, the potential of potassium and tetraphenylphosphonium ion-selective electrodes changed, indicating an increase in cytoplasmic and outer membrane permeability, respectively. No changes in membrane permeability were observed in cells treated with capric acid or monocaprin. Transmission electron microscopy revealed separation of the inner and outer membrane in cells treated with capric and fumaric acid, as well as cytoplasmic disorganization in cells exposed to capric acid.

Phytase activity in rabbit cecal bacteria.

The presence of phytase activity was demonstrated in 26 strains of rabbit cecal bacteria. In 25 strains a low phytase activity, 0.10-0.62 micromol phosphate released per min per mg protein, was found. High activity (2.61 micromol/min per mg protein) was found in the strain PP2 identified as Enterococcus hirae. Phytase activity was cell-associated, being higher in the cell extract than in the cell walls. Extracellular phytase activity and cell-associated phosphatase activity were not detected. Phytase activity was optimal around pH 5.0, which is below the physiological cecal pH range. The K (m) determined using the Lineweaver-Burk plot was 0.19 micromol/mL. Cations Fe(3+), Cu(2+) and Zn(2+) at 0.5 mmol/L decreased phytase activity in sonicated cells of E. hirae by 99.4, 90.7 and 96.5 %, respectively. In contrast, Mg(2+) increased activity by 11.0 %. Characteristics of E. hirae phytase (pH optimum, K (m), cation sensitivity) were similar to those of other bacterial phytases reported in the literature. Other bacteria with a high phytase activity may be present in the rabbit cecum but remain to be identified.

Dietary selenium increases vitamin E contents of egg yolk and chicken meat.

1. The influence of different forms of dietary selenium (Se) on vitamin E (alpha-tocopherol) and Se contents of egg yolk and chicken meat was investigated. 2. Eggs were collected from laying hens subjected to 4 different dietary treatments after 24 weeks of age. Treatments compared the effects of inorganic Se supplementation (selenite) to those of organic (Se-enriched yeast, Se-enriched alga Chlorella) supplements. In a second experiment the effect of the above organic dietary Se supplementation on the alpha-tocopherol contents of meat from broiler chickens was evaluated. 3. Dietary Se supplementation increased the alpha-tocopherol content of egg yolks from 297 mg/kg dry matter in treatment without supplementation to 311 mg/kg when selenium was supplemented as selenite, and to 370-375 mg/kg when organic supplements were used. The Se and alpha-tocopherol contents of breast and thigh meat in broilers were significantly increased by organic dietary Se supplementation. 4. The inclusion of organic dietary Se sources in the diets of laying hens and broilers would enhance the nutritional value (vitamin E and Se contents) of products (eggs and meat) for human consumption.

Influence of pH on antimicrobial activity of organic acids against rabbit enteropathogenic strain of Escherichia coli.

Susceptibility of the rabbit enteropathogenic strain Escherichia coli C6 (O128 serogroup) to C6-C14 fatty acids, oleic, citric, lactic and fumaric acid at 5 mg/mL was determined by the plating technique in the near-neutral pH region (pH approximately 6.5), and in a weakly acid and acid environment (pH 5.4 +/- 0.1 and 2.2-2.5, respectively). In the near-neutral pH region caproic and caprylic acid reduced the concentration of viable cells by 3 and 6 orders, respectively. At lower pH the bactericidal effect of caproic acid remained similar, but caprylic acid decreased the concentration of viable cells to < 100/mL. The bactericidal activity of capric acid was low at pH 6.5 but increased at pH 5.3. High environmental acidity was intrinsically bactericidal and at very low pH the effects of fatty acids were thus less pronounced. Citric acid reduced the counts of viable cells to 1/10. Antimicrobial activity of other acids examined was marginal or absent. Medium-chain fatty acids, caprylic and, to a lesser extent, also caproic and capric acid were better antimicrobials than other organic acids examined; the antimicrobial activity of fatty acids toward the C6 strain was pH-dependent. Beneficial effects of citric, lactic and fumaric acid reported by animal nutritionists are thus probably related to factors other than their direct antimicrobial action.

Effect of pectin and amidated pectin on cholesterol homeostasis and cecal metabolism in rats fed a high-cholesterol diet.

Two experiments were performed to compare the effect of pectin and its hydrophobic derivatives on homeostasis of cholesterol and cecal metabolism in male young rats. Control rats were fed a diet supplemented with palm fat and cholesterol (50 and 10 g/kg, respectively). Rats of other groups were fed the same diet containing citrus pectin or octadecylpectinamide (60 g/kg). Diets were fed for 4 weeks. In experiment I, pectinamide of lower degree of amidation (30 %) increased serum HDL cholesterol from 1.20 to 1.43 micromol/ml (p>0.05) at the expense of other cholesterol fractions. In experiment II, pectinamide of a higher degree of amidation (53 %) significantly decreased total serum cholesterol from 2.08 to 1.67 micromol/ml. Amidated pectins at both levels of substitution significantly decreased hepatic concentrations of cholesterol and fat. In both experiments the relative weight of cecum in the pectinamide group was significantly lower than in pectin group. The highest cecal concentrations of short-chain fatty acids (SCFA) were found in rats fed a diet with pectin (133.2 and 129.3 micromol/g in experiment I and II, respectively). In other groups, cecal SCFA was significantly (pectinamide groups) or non-significantly (controls) lower. In wet feces, SCFA concentrations were higher and butyrate molar proportions lower than in corresponding cecal contents. Pectinamide of a lower or higher degree of substitution significantly increased fecal content of cholesterol from 18.5 and 17.3 micromol/g in controls to 31.8 and 28.0 micromol/g, respectively. Corresponding concentrations of coprostanol were decreased. Effects of pectin on cholesterol homeostasis were absent or marginal. Histological examination revealed that hepatic tissue of control and pectin-fed rats was infiltrated with lipids. The Sudan black-positive material was absent in the liver of rats fed pectinamides. No pathological changes of liver tissue were apparent. In summary, hydrophobic amidated pectins significantly altered cholesterol homeostasis in rats and might be considered as a clinically effective hypocholesterolemic agent. Low cecal SCFA concentrations in rats fed pectinamides suggest that amidation of pectin had decreased its fermentability.

Influence of dietary selenium and vitamin E on quality of veal.

Three groups of six calves each were fed a milk replacer and a starter concentrate for 15weeks. Calves of the first group received the basal diet containing selenium (Se) and vitamin E at 0.095-0.128mg and 30-33mg per kg of total solids, respectively. Calves of the second group received the basal diet supplemented with Se-enriched yeast to increase dietary Se concentration to 0.50mg/kg. The third group of calves received the latter diet supplemented with vitamin E to increase its concentration to 100mg/kg. There was no effect of diet on growth rate, digestibility of dry matter and Se, chemical composition of meat (M. Longissimus thoracis et lumborum), meat colour and fatty acid profile of meat lipids. The Se supplementation significantly increased Se concentration in muscle from 0.21 to 0.43mg/kg. The activity of glutathione peroxidase (GSH-Px) in muscle and liver tissue of Se-supplemented animals was increased by 56% and 67%, respectively, compared to the control. The combined supplementation of vitamin E and Se significantly improved the lipid stability of meat compared to the control diet, but not compared to the Se-supplemented diet. It can be concluded that dietary Se supplementation increases the concentration of Se and the GSH-Px activity in meat, but has limited potential for improving meat oxidative stability.

Growth performance and fatty acid profiles of intramuscular and subcutaneous fat from Limousin and Charolais heifers fed extruded linseed.

Forty-eight heifers, Limousin (LI) and Charolais (CH) breed, were used to evaluate the effect of a basal mixed diet with linseed supplementation (108g/kg DM) on performance and fatty acid (FA) composition of M. longissimus thoracis (MLT) and subcutaneous fat. The feed intake and weight gains were higher, and feed per gain ratio lower in CH heifers than in LI heifers. The muscle lipids and subcutaneous fat of LI heifers contained significantly more MUFA, PUFA, CLA, and less SFA than the lipids of CH heifers. The feeding of extruded linseed significantly increased the concentration of linolenic acid (C18:3n-3), CLA, and decreased PUFA n-6 to PUFA n-3 ratio in lipids extracted from both tissues. Linseed supplementation decreased SFA and increased PUFA in subcutaneous fat. In MLT lipids of linseed-fed heifers PUFA of the n-3 series were increased. It can be concluded that the growth performance of LI heifers was lower, but that their lipids contained more FA that are thought to be important for human health. The feeding of linseed at 108g/kg DM enhanced the nutritive value of beef in terms of FA profile. Breed differences were generally more important than effects of the diet. Dietary effects, however, were more pronounced in PUFA n-3 and arachidonic acid (C20:4n-6).

Effect of a starter diet supplementation with mannan-oligosaccharide or inulin on health status, caecal metabolism, digestibility of nutrients and growth of early weaned rabbits.

The effect of a dietary supplementation with mannan-oligosaccharide (MOS, Bio-Mos, Alltech Inc.) and inulin (Frutafit® IQ) on growth, health, and caecal traits was studied on 348 rabbits (Hyplus®), weaned at 25 days of age. Three hundred and thirty rabbits (110 per group) were used for the health status and growth performance trial, while 18 rabbits (six per group) were used for caecal metabolism evaluation at the age of 42 days of age. Three diets were formulated: C (control), M (0.3% MOS) and I diet (4% inulin). Digestibility of the diets was measured in 10 rabbits per group between 36 and 40 days of age. The control diet was fed to rabbits of the C group from weaning to 74 days of age (slaughter). Diets M and I were fed to rabbits of the respective group from weaning to 46 days of age, then were fed with control diet till slaughter. From 25 to 46 days of age, the weight gain was slightly higher in control rabbits ( P = 0.11), while no differences were recorded for the whole period. No differences among groups in the mortality, which was high due to an enteropathy-infected environment, were significant. The lowest morbidity ( P = 0.05) as well as the health risk index were recorded in rabbits fed the diet with inulin ( P = 0.03). After change of diet, the health risk index increased in the rabbits previously fed the diet with additives, thereby no significant differences in the health status were recorded for the whole period. Total caecal volatile fatty acids concentration was higher ( P < 0.01) and the pH ( P < 0.01) and ammonia concentration ( P = 0.01) lower in rabbits fed the inulin diet than in other rabbits. In these animals, acetate molar proportion was higher ( P = 0.01) and that of propionate as well as the propionate/butyrate ratio significantly lower than in other rabbits. Butyrate molar proportion was higher in rabbits fed the diet with MOS ( P < 0.01). In rabbits fed the inulin diet a higher activity of inulinase was recorded ( P < 0.001) than in other rabbits. A significantly lower digestibility of cellulose was observed in rabbits fed the diet with MOS. The results of our study suggest the importance of using inulin-type fructans in the nutrition of young rabbits. The higher health risk index of rabbits after change of diets indicates that prebiotics should be given for a longer time during the fattening period.

Activity and cellular localization of amylases of rabbit cecal bacteria.

Five 11-week-old rabbits, fed a commercial granulated feed, were slaughtered and cecal starch-degrading bacteria enumerated; total concentration of cultivable bacteria utilizing starch averaged 5.5 x 10(10) CFU/g. The activity and cellular localization of amylases was determined in 9 bacteria identified as Actinomyces israeli (strains AA2 and AD4), Bacteroides spp. (strain AA3), Dichelobacter nodosus (strain AA4), Mitsuokella multiacidus (strain AA6), Eubacterium spp. (strains AA7 and AB2), Clostridium spp. (strains AD1 and AA5). Four strains (AA3, AA4, AA5, AD4) produced extracellular amylases with an activity of 26-35 micromol of reducing sugars per h per mg of protein; in five strains (AA2, AA6, AA7, AB2, AD1) amylases were membrane-bound with an activity of 14-18 micromol of reducing sugars per h per mg of protein. All strains exhibited a low intracellular amylolytic activity. The pH optimum of amylases was 6.8-7.0. In strains producing extracellular amylases a substantial loss of viscosity was observed during incubations of cultivation supernatant with starch, similar to viscosity reduction in starch solutions treated with alpha-amylase; this indicates an endo-type (random cleavage) of extracellular amylase reaction in the bacteria under study. No strain possessed glucoamylase activity.

Different utilization of glucose and raffinose in Bifidobacterium breve and Bifidobacterium animalis.

Growth of 2 strains of Bifidobacterium breve and 4 strains of Bifidobacterium animalis was evaluated with glucose, raffinose or a mixture of glucose and raffinose as substrates. All strains of B. animalis and one strain of B. breve grew more slowly on glucose than on raffinose or the mixture of glucose and raffinose; one strain of B. breve grew more rapidly on the mixture of glucose and raffinose than on raffinose while 2 strains of B. animalis grew faster on raffinose than on the mixture of raffinose and glucose. Both strains of B. breve utilized glucose and raffinose simultaneously. In contrast, all strains of B. animalis strains displayed atypical growth with rapid utilization of raffinose followed by slow utilization of glucose. The cell morphology of all strains of B. animalis was affected by the sugar used for cultivation: tiny and rather spherical cells were observed on glucose while on media with raffinose the cells were large and had the species-specific shape. Description of preferential utilization of various saccharides by bifidobacteria can contribute to the development of new synbiotic preparations and new cultivation media for bifidobacteria.

Effect of various copper supplements to feed of laying hens on cu content in eggs, liver, excreta, soil, and herbage.

Copper is often added to poultry diets as an antimicrobial agent at doses greatly exceeding the nutritional requirement. In this study, the basal diet of laying hens containing 9.2 mg Cu/kg was supplemented with CuSO(4) x 5H(2)O at 0, 25, 65, 115, and 240 mg Cu/kg dry matter (DM). At Cu dietary concentration just below the level permitted by the European Union (35 mg/kg), the Cu content in the egg yolk was significantly (p < 0.05) increased by 26%, and nonsignificantly by 4.1, 9.1, and 7.9% in the egg white, eggshell, and liver, respectively. When Cu concentration in the diet was doubled, the effect of Cu on Cu content in eggshell and liver was statistically significant as well. In no liver sample was the hygienic limit of Cu content (80 mg/kg) exceeded. Supplementation of diets with Cu increased Cu concentration in excreta linearly from 25.3 to 396.8 mg/kg DM. Dried excreta were used for fertilization of grassland at 21 g N/m(2). Three months later, soil and herbage were sampled and analyzed. The Cu concentration in soil increased from 25.3 to only 46.4 mg/kg DM when dietary Cu concentration rose from 9.2 to 243.7 mg Cu/kg DM. Corresponding Cu concentrations in herbage were 6.8 and 19.2 mg/kg DM. It can be concluded that the deposition of Cu in eggs and liver of hens fed Cu-supplemented diets does not represent a hygienic risk. The accumulation of Cu in soil fertilized with excreta of Cu-fed hens and in herbage was limited.

[Inhibition of carbohydrate metabilsm enzymes by pentachlorophenol in cells of pectinolytic bacteria from gastrointestinal tract of animals and effect of clinoptilolite adsorbents on this process].

Changes in functional activity of specific enzyme reactions in the cells of pectinolytic bacteria from the gastrointestinal tract of animals in vitro cultivated in the medium containing pectin or glucose were studied against a background of the low dose effect of the wide spread biocide pentachlorophenol alone as well as in combination with the natural sorbents clinoptilolites. Regardless of the absence of transketolase reaction in the cells of all studied strains, they metabolized highly the above substrates that are dissimilar in chemical structure and produced different products of their degradation. It has been shown that the high metabolic level in the cells is provided by the function of the unique enzymatic reaction catalyzed by 2-keto-3-desoxy-6-phosphogluconate aldolase (EC 4.1.2.14) that permits to use effectively the metabolic pathway of Entner-Doudoroff. Cells could also utilize the same substrates via the Embden-Meyerhof-Parnas pathway, therefore they possess the other key reaction that is catalyzed by fructosobiphosphate aldolase (EC 4.1.2.13). Even a low dose of PCP (20 microM) decreased sharply activity of the mentioned key enzymes and intermediates' production in the cells of the studied strains with the use of both substrates. However, presence of clinoptilolites in the medium reduced significantly the biocide inhibition effect. Furthermore, in the medium with glucose, protection of intracellular metabolism with the help of sorbents was registered more clearly than with pectin. This can evidence for more mobile and simpler possibilities of accelerated production of necessary intermediates from glucose that are capable to induce activation of the key enzymatic reactions in cells utilizing selectively the substrates (which are different in accessibility and other characteristics) under the toxic agent effect.

Effects of dietary zinc, iron, and copper in layer feed on distribution of these elements in eggs, liver, excreta, soil, and herbage.

An experiment was conducted to evaluate the effect of dietary content and combinations of Zn, Fe, and Cu on deposition of these elements in egg components, liver, and excreta. Excreta were applied as a manure to a lawn, and 3 mo later soil and herbage samples were taken and analyzed. The experiment comprised 144 hens in 8 groups. The basal diet contained Zn, Fe, and Cu at 63.4, 92.8, and 9.0 mg/kg, respectively. It was supplemented with 1, 2, or 3 trace elements (inorganic forms) at 80 mg of Zn/kg, 120 mg of Fe/kg, and 25 mg of Cu/kg. Recovery of Zn, Fe, and Cu in eggs of hens fed the basal diet was 10.7, 9.8, and 4.4% of the alimentary intake, respectively. A Zn-Cu antagonism was observed; deposition of Zn in the yolk was significantly decreased by Cu addition and vice versa (P < 0.01). Supplementation of the basal diet with Fe increased Fe concentration in egg yolk and white by 6.3 and 2.2%, respectively. The combination of Fe with Zn and Cu, however, increased Fe concentration in the yolk and white by 36.7 and 34.9%, respectively (P < 0.01). The enrichment of eggs with the other elements was marginal (Cu) or absent (Zn). Effects of Zn, Fe, and Cu of the basal diet on liver concentrations of these elements were relatively small, and no antagonism between Zn and Cu was apparent. Supplementation of the basal diet with the combination of Zn and Fe, however, significantly decreased hepatic concentration of Cu. On the other hand, Cu supplementation significantly increased Fe concentration in livers of hens fed the Fe-supplemented diet (P < 0.01). Concentrations of Zn, Fe, and Cu in excreta were related to their dietary content. High concentrations of Zn, Fe, and Cu in excreta corresponded with limited deposition of the 3 elements in eggs and liver. Concentrations of Zn, Fe, and Cu in herbage correlated significantly with the supply of these elements by hen excreta into soil. The Zn supplied by hen excreta was more stable than Fe and Cu; thus Zn could accumulate in the soil.