S. Smulikowska

Effects of sodium butyrate and salinomycin upon intestinal microbiota, mucosal morphology and performance of broiler chickens.

The effect of dietary sodium butyrate (SB) or salinomycin (SAL) or both additives on performance, small intestinal morphology and microbial ecology of broiler chickens was studied. A growth trial was conducted with 96 Ross 308 female broilers from 1 to 30 days of age. Four treatment groups were fed with a non-supplemented control diet or three experimental diets supplemented with i) 300 mg SB (Adimix 30 coated) per kg, ii) 60 mg SAL (Sacox) per kg or iii) both additives in combination. Feed intake and body-weight gain decreased and gain-to-feed ratio increased due to SAL supplementation, while addition of SB did not affect performance in comparison with the control diet but positively affected feed intake and body-weight gain in comparison with birds fed the SAL-supplemented diet. Villus height in jejunum decreased, while crypt depth increased due to SAL supplementation. Addition of SB increased crypt depth in jejunum. No significant effect of either additive was observed in ileum morphology. Total short-chain organic acids concentration in ileal digesta decreased with SAL supplementation, mainly due to lower lactic acid concentration, but no effects were observed in the caeca. The SAL supplementation was accompanied by a pH increase in ileum and a pH decrease in caecum. No significant effect of SB addition was observed for these parameters. Total bacterial numbers and Lactobacillus [lactic acid bacteria (LAB)] counts in ileal and caecal contents were lower in birds fed with SAL-supplemented diet in comparison with birds fed with control or SB diet. DNA fingerprints revealed SAL supplementation to affect the microbial population by suppressing dominating LAB, potentially L. aviarius. The presented results show that dietary SAL, supplemented alone or in combination with SB, suppressed the microbial activity and altered the microbial community structure mainly in ileum. SAL alone negatively affected feed intake and body-weight gain; however, the effect was ameliorated by SB supplementation.

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.

Versuche zur Bestimmung des endogenen und exogenen fäkalen N-Anteiles monogastrischer Tierarten

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.

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.

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 0.3g/kg diet for rats, 0.65 g for chicken and at least 0.5 g for growing pigs.

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

&NA; 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.

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.

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

ABSTRACT 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.

Probiotic preparation reduces faecal water genotoxicity and cytotoxicity in chickens fed ochratoxin A contaminated feed (in vivo study)

The aim of the present study was to assess the genotoxicity and cytotoxicity of the faecal water of chickens fed ochratoxin A (OTA) contaminated feed with and without probiotic preparation. The study was performed on 20 healthy female Ross broiler chickens divided into 4 groups: control chickens - fed with non-supplemented feed; PP chickens - fed feed supplemented with the probiotic preparation; OTA chickens - fed feed contaminated with 1 mg per kg of OTA; OTA + PP chickens - fed feed contaminated with 1 mg per kg of OTA and supplemented with the probiotic preparation. Faecal water samples were collected on the 35(th) day of life of chickens from each group. Genotoxicity was measured using the comet assay, and cytotoxicity by means of MTT tests. Mean DNA damage, measured as the percentage of DNA in the tails of the comets, was 8.50 ± 1.10 for chickens fed OTA at 1 mg/kg and 6.41 ± 0.67 in the controls. The supplementation of feed with the probiotic preparation decreased the extent of DNA damage to 4.74 ± 0.78. In the control group of chickens the average cytotoxicity was 38.5 ± 0.5 (in MTT), while in the probiotic preparation group (PP group) it was 31.8 ± 0.7 (in MTT). After supplementation of the feed with the probiotic preparation, the genotoxicity and cytotoxicity were decreased in a statistically significant manner.

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

ABSTRACT 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 β-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. β-glucosidase, α-galactosidase and β-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.