Effects of pea extrusion and enzyme and probiotic supplementation on performance, microbiota activity and biofilm formation in the broiler gastrointestinal tract.

1. The effects of supplementation of broiler chicken diets with pea meal, carbohydrase enzymes and a probiotic were investigated for potential performance improvement. 2. Raw or extruded pea meal (cv Model, grown in Poland) was included in a wheat-soybean meal-based diet at 250 g/kg. The diets were unsupplemented (control) or supplemented with either carbohydrase enzymes (200 U/kg xylanase and 10 U/kg ß-glucanase in feed) or a probiotic (Bacillus subtilis), or both. The diets were fed to Ross 308 broilers aged 9-28 days. 3. After two additional days, chick gastrointestinal tracts were excised and analysed for the presence of Bacillus subtilis biofilm; and the ileal and caecal digesta were analysed for bacterial enzyme activities and to determine the concentration of short-chain fatty acids (SCFAs). 4. Feeding the pea-based diet supplemented with the probiotic compromised feed utilisation, due to higher feed intake. The addition of enzymes to the raw, but not the extruded, pea containing diet partially ameliorated this effect (pea form × additives; P < 0.002). 5. In the ileal digesta, interactions between the dietary treatments were observed for the activities of all bacterial glycolytic enzymes and for SCFA concentrations. ß-glucosidase, α-galactosidase and ß-glucuronidase were highest in birds fed the diet containing extruded pea supplemented with the probiotic and enzymes (pea form x additives; P = 0.018 to P < 0.006). In the caecal digesta, interactions were observed for bacterial enzyme activities, but not for total SCFA concentration. Biofilm formation in the caecum indicated that the probiotic strain was metabolically active in the broiler gut. 6. In conclusion, supplementation of diets containing raw or extruded pea meal with enzymes and a Bacillus subtilis spore-based probiotic modulated microbiota activity but had no clear effects on broiler performance. Probiotic administration did not cause excessive fermentation in the ileum and caecum but enhanced Bacillus subtilis spp. biofilm formation in the caecum, which may be indicative of a beneficial effect on gut health.

The interactive effect of dietary n-6: n-3 fatty acid ratio and vitamin E level on tissue lipid peroxidation, DNA damage in intestinal epithelial cells, and gut morphology in chickens of different ages.

Feeding chickens diets high in n-3 fatty acids (FA) increases their incorporation into tissue lipids, but leads to oxidative stress in cells. This study investigated the effect of the dietary polyunsaturated FA ratio (PUFA n-6: n-3) and vitamin E (vE) level on DNA damage and morphological changes in the gut epithelium of chickens. One-day-old female broiler chicks (n = 176) were divided into 4 groups fed for 43 d diets with a high (HR) or low (LR) PUFA n-6: n-3 ratio and supplemented with 50 or 300 mg vE kg-1. Performance was calculated for periods of d 1 to 9, d 9 to 16, d 9 to 35, and d 9 to 42, while organs were sampled at d 9, d 17, d 36, and d 43. At d 17 and d 43, DNA damage of epithelial cells in the duodenum and jejunum was measured and duodenal and jejunal morphology was analyzed. HR diets improved FCR for the periods of d 1 to 9, d 9 to 16 and d 9 to 42, whereas the increased vE level improved FCR for the period of d 9 to 16. In the jejunum DNA damage was greater in chickens fed LR than HR diets at d 17 (P < 0.001) and the increased vE level promoted DNA damage in both intestinal segments (P < 0.02) in younger birds. The morphology of the duodenum was marginally affected by the diets, whereas LR diets in the jejunum reduced villus surface area at d 17 (P = 0.022), and mucosa thickness (P = 0.029) and villus height (P = 0.035) at d 43. The results indicated that feeding birds LR diets and vE levels significantly exceeding the recommendation induced DNA damage in epithelial cells, but this effect varied depending on the intestinal segment and the age of birds.

Viscosity negatively affects the nutritional value of blue lupin seeds for broilers.

This study examines the impact of Lupinus angustifolius variety (C) and inclusion level (L) in broiler diets on the nutritional value, viscosity of ileal digesta and activity of gut microbiota. The experiment was conducted on 154 female 21-day-old broilers, allocated to 11 groups (kept individually). A reference lupin-free diet and 10 test diets containing one of five lupin seeds; Kadryl, Regent, Dalbor, Bojar and Tango, mixed with the reference diet at a ratio of 25 : 75 or 32 : 68 dry matter (DM) (low or high level of inclusion) were prepared. Diets were fed for 6 days, excreta were collected over last 4 days. Apparent metabolizable energy corrected to zero N balance (AMEN) of diets and AMEN of lupin seeds were calculated. Birds were sacrificed, ileal and caecal digesta were pooled by segments from two birds, and the activity of bacterial enzymes was determined. The ileal digesta viscosity was measured immediately (ileal viscosity immediate (IVI)) or after 6 days storage at -18°C (ileal viscosity frozen). AMEN of test diets were lower than the reference diet. Lupin AMEN values ranged from 6.04 MJ/kg DM for Regent at high level to 9.25 MJ/kg DM for Bojar at low level. High inclusion level numerically decreased AMEN value in all cultivars, except for Kadryl, for which it increased (significant C×L interaction). The IVI value was 2.6 mPa·s in the reference group, but ranged from 6.3 to 21.7 mPa·s in lupin-fed birds. It increased significantly with level for Regent, Dalbor and Tango but not for the other two cultivars (significant C×L interaction). There was a negative correlation between IVI and: apparent total tract N retention, fat digestibility from test diets, AMEN of diets and lupins. Ileal viscosity immediate was positively correlated with the activity of ileal α- and ß-glucosidase and negatively with ileal α-galactosidase and caecal α-glucosidase. Ileal viscosity frozen ranged from 3.2 to 5 mPa·s and it was not correlated with lupins AMEN. This suggests that the digesta viscosity caused by narrow-leafed lupin is detrimental to its nutritional value and interfere with the gut microbial activity. In addition, the lupins viscosity was measured by two in vitro methods: the water extract viscosity (WEV) method and after incubation in conditions imitating in vivo digestion (enzyme-treated extract viscosity (EEV)). In vivo viscosity was weakly reflected by in vitro measurements as there was no correlation between IVI and WEV or EEV. Overall, findings suggest that the different cultivars of narrow-leafed lupin may have different value for practical application in broiler diets.

Beneficial effects of enrichment of chicken meat with n-3 polyunsaturated fatty acids, vitamin E and selenium on health parameters: a study on male rats.

Consumption of chicken meat enriched with bioactive compounds such as n-3 polyunsaturated fatty acids (PUFAn-3), vitamin E (vE) and selenium (Se) can help prevent many diseases and can be used to deliver those substances to humans. This might be of importance as chicken meat consumption is increasing worldwide. The effects of enriching chicken meat with PUFAn-3, vE and Se through dietary interventions were studied in rats. Four groups of Ross 308 female broilers from day 22 to day 35 of age were fed control diet (L) that contained lard and 80 mg vE and 0.3 mg Se/kg, or diets that contained rape seeds and fish oil with the same level of Se and vE as in the control diet, the same level of Se as in the control and 150 mg vE/kg, or 150 mg of vE and 0.7 mg Se/kg. Broiler carcasses were boiled, deboned, lyophilized and pooled by group. Boiled edible components of chicken carcass (BECC) were included (240 g/kg) in the diets fed to four groups of ten 10-week-old Wistar male rats for 8 weeks. Inclusion of BECCs modulated dietary fatty acid profile in the rat diets. Feeding these diets did not influence parameters related to growth or relative weights of internal organs in the rats. Feeding BECCs with lower PUFAn-6/n-3 decreased the n-6/n-3 ratio in the rat brain and liver, and increased the proportion of docosahexaenoic acid in the brain lipids. Liver cholesterol level was similar among the experimental groups, whereas the concentration of vE in the liver of rats fed BECC with increased vE levels was higher than that in the rats fed BECC with the basal vE level. Haematological and biochemical parameters in blood were within the normal range for rats, but a few rats showed a tendency towards increased levels because of the higher vE and Se level. The health-promoting effect of feeding rats PUFAn-3 enriched BECC was more pronounced when an increased dietary level of vE was used, but the increased level of Se did not provide the rats with additional benefits. Thus, the findings indicate that BECC enriched with PUFAn-3 and vE by a dietary intervention is a functional food with great potential of implementation.

Interactive effects of dietary lipids and vitamin E level on performance, blood eicosanoids, and response to mitogen stimulation in broiler chickens of different ages.

The effects of the dietary polyunsaturated fatty acids (PUFA) n-6:n-3 ratio and vitamin E (vE) on the levels of pro-inflammatory eicosanoids, the incorporation of docosahexaenoic acid (DHA) and arachidonic acid (AA) into immune tissues, and changes in leukocyte population after phytohemagglutinin (PHA) challenge were investigated in broiler chickens of different ages. One-day-old female broilers (48 per treatment) were fed 4 different wheat-soybean-corn-based diets containing corn oil with a high PUFA n-6:n-3 ratio (HR) or a mixture of linseed and fish oils with a low PUFA n-6:n-3 ratio (LR). Diets contained either 50 mg vE kg-1 of diet (basal vE) or 300 mg vE kg-1 of diet (increased vE). At d 14 and d 34, 8 chickens per treatment were challenged with PHA, and wing web swelling (WWS) was measured. The blood concentration of leukotriene (LTB4), prostaglandin (PGE2), and thromboxane (TBX2) in 17-day-old and 43-day-old chickens was determined. The pattern of AA and DHA incorporation into bursa, spleen, and brain lipids reflected the level of their precursors in the diet. WWS was the highest in chickens fed a LR diet and in 14-day-old chickens (P < 0.01). Leukocyte proportions varied with dietary PUFA n-6:n-3 ratio and with age. The heterophil:lymphocyte ratio was the highest at 6 h post PHA challenge, and was higher in 34-day-old chickens (P < 0.001). TBX2 and PGE2 concentrations were higher in chickens fed HR diet, whereas TBX2 and LTB4 concentrations were lower at high vE level. Lower PGE2 and LTB4, but higher TBX2 concentrations were measured in younger birds (P < 0.001). The results indicated that LR increased the phagocytic cell proportion in the blood; HR promoted the incorporation of AA into the immune tissues, which increased the levels of more pro-inflammatory eicosanoids in the blood; and vE counteracts these effects to some extent. Owing to the immaturity of the immune system, dietary interventions might be promising at the early stage of chicken growth.

Influence of dietary peas and organic acids and probiotic supplementation on performance and caecal microbial ecology of broiler chickens.

1. The effect of dietary pea and addition of organic acid blend (OA) or probiotic (Pro) on performance and caecal microbial ecology of broiler chickens was studied. 2. A growth trial was conducted with 160 Ross 308 female broilers from d 1 to 35 of age. There were 8 treatment groups based on either control (S) or white pea (P). Both S and P were supplemented with OA (Galliacid - fumaric acid, calcium formate, calcium propionate and potassium sorbate coated with plant triglycerides, Vetagro) and or with Pro (LABYuc-Probio - lactic acid bacteria, Saccharomyces cerevisiae and Yucca schidigeri extract, Mifarmex GmbH). 3. Inclusion of peas in the diet increased feed intake and decreased gain:feed ratio in comparison to the control diet. Neither probiotic nor OA supplementations affected broiler performance. 4. The caecal microbiota was characterised in 37-d-old birds by fluorescent in situ hybridisation (FISH) and terminal-restriction fragment length polymorphism (T-RFLP). Total bacterial counts in caecal contents were slightly higher for birds fed the pea diets, but were not affected by OA or Pro supplements. 5. Neither pea nor Pro affected the Lactobacillus/Enterococcus and Streptococcus/Lactococcus counts in caecal contents, whereas OA supplementation slightly increased the Lactobacillus/Enterococcus counts. The composition of the Lactobacillus/Enterococcus population was altered by inclusion of peas as revealed by the T-RFLP patterns. 6. The DNA fingerprint further suggested that the caecal microbiota was dominated by the lactic acid bacterium Streptococcus alactolyticus. 7. In ileal contents, the concentration of short-chain fatty acids (SCFA) was decreased only by Pro supplementation. In caecal contents, the SCFA concentration was higher for birds fed on the pea diets, and increased significantly with Pro supplementation 8. In conclusion, the results indicate that the use of pea and probiotics in broiler feed may stimulate the caecal commensal microbiota (growth and/or activity) to some extent and hence prevent establishment of pathogenic and zoonotic enterobacteria in these segments of the gut.

Effect of an organic acid blend and phytase added to a rapeseed cake-containing diet on performance, intestinal morphology, caecal microflora activity and thyroid status of broiler chickens.

The experiment was carried out on 96 female broilers, allocated to eight groups of 12 birds kept in individual cages. Two basal wheat- and soyabean meal-based diets containing 150 g/kg of rapeseed expeller cake were formulated, differing in the level of P: 7.1 g/kg in diet H or 5.9 g/kg in diet L. Rapeseed cake supplied 3.15 micromol alkenyl glucosinolates per gram of diet. The eight treatments were: basal diets only, basal diets + phytase (1000 U/kg), basal diets + organic acid blend (OA, 6 g/kg), or basal diets + both additives. Diets were fed from day 8 to 28 of life. The results showed that the lower dietary P content and OA supplementation did not significantly affect feed intake or BWG, while both increased (p < 0.001) after phytase supplementation. Tibia ash content as well as tibia ultimate strength were lower (p < 0.001) in birds fed diets L compared with diets H, and increased (p < 0.01) with phytase supplementation of diet L, while OA had no influence on either parameter. Dietary P levels and OA supplementation had no influence on the pH of gut digesta, but the pH of jejunal digesta increased following phytase supplementation (p < 0.01). Morphological measurements of the small intestinal mucosa of chicks indicated that OA added to diet L depressed villi height (p < 0.001) and crypt depth (p < 0.001); both parameters increased after phytase supplementation (p < 0.01). The lower total SCFA as well as acetic, propionic and butyric acid concentrations in caecal digesta indicated lower activity of caecal microflora in birds fed diets L compared with H. OA supplementation had no influence, while phytase supplementation increased the concentration of acetic acid in caecal digesta. Supplementation of diets with either phytase or OA increased thyroid weight by 16% (p < 0.01) and 11% (p < 0.05) respectively. The increase in thyroid weight because of phytase supplementation was greater at the lower dietary P level, and the greatest when both phytase and OA were added to the diet.

Response of animals to dietary gramine. I. Performance and selected hematological, biochemical and histological parameters in growing chicken, rats and pigs.

The effects of feeding varied levels of low- and high-gramine yellow lupin seeds (LG and HG, respectively), and of synthetic gramine added to the diets in amounts ranging from 0.15 to 1.2 g per kg were investigated in one experiment on growing chicken and in two experiments on growing rats. The comparison of LG and HG lupin and the effect of 0.5 g gramine per kg of LG diet were determined in a growth-balance experiment with pigs. Organ weights and histology, blood parameters and activity of liver enzymes were determined. The response to HG lupin and gramine concentration varied among the species, the rats being more affected than chicken; no adverse effects of HG lupin or gramine were found in growing pigs. The common reaction of rats and chicken to the high levels of gramine (native or synthetic) was the decrease of feed intake and body gain. The increase of the relative weight of liver or kidney, changes in hematological parameters and liver enzymes were found only in rats. The estimated NOAEL (no-observed-adverse-effect level) of gramine was about 0.3 g/kg diet for rats, 0.65 g for chicken and at least 0.5 g for growing pigs.

[Determination of endogenous nitrogen in feces using the isotope technic]. / Zur Bestimmung des endogenen Stickstoffs im Kot mit Hilfe der Isotopentechnik.

A ration consisting of wheat gluten and N-free components was supplemented with L-lysine and L-leucine and fed to two groups of growing Wistar rats. Group 1 received 15N Lys and unlabelled Leu, group 2 received unlabelled Lys and 15N Leu in order to study the influence of the utilization of the 15N marker on the labelling quota of faeces and urine as well as various fractions of the body. The good utilization of Lys in group 1 results in a higher 15N excess in faeces and a reduced 15N abundance in urine in comparison to group 2 with a lower utilization of 15N Leu. The results show that the 15N abundance in urine is unsuitable as an indicator of the 15N labelling quota of endogenous metabolic faecal nitrogen.

[Attempts at the determination of endogenous and exogenous fecal proportions in monogastric animals]. / Versuche zur Bestimmung des endogenen und exogenen fäkalen N-Anteiles monogastrischer Tierarten. 1. Einleitung

A trial was made with growing Wistar rats receivine 15N labelled tracers. The animals were fed maize grains (Opaque). Subsequently, the level of urinary and faecal N excretion was measured and the 15N frequency was determined in the urine, faeces and different blood fractions. On the basis of the model established by Czarnetzki for the nitrogen metabolism in monogastric animals the estimated values were used to calculate the proportion of endogenic faecal N in the total amount of faecal nitrogen. In agreement with previous trails it was found that increase in protein intake produced a significant increase in endogenic faecal N; similarly, a higher value was found for the true digestibility of maize protein (by 9.3 units) compared with that established by the conventional regression technique.