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The potential of a phytogenic premix (PP) based on ginger, lemon balm, oregano and thyme to stimulate the expression of cytoprotective genes at the broiler gut level was evaluated in this study. In particular, the effects of PP inclusion levels on a selection of genes related to host protection against oxidation (catalase [CAT], superoxide dismutase 1 [SOD1], glutathione peroxidase 2 [GPX2], heme oxygenase 1 [HMOX1], NAD(P)H quinone dehydrogenase 1 [NQO1], nuclear factor (erythroid-derived 2)-like 2 [Nrf2] and kelch like ECH associated protein 1 [Keap1]), stress (heat shock 70 kDa protein 2 [HSP70] and heat shock protein 90 alpha family class A member 1 [HSP90]) and inflammation (nuclear factor kappa B subunit 1 [NF-κB1], Toll-like receptor 2 family member B (TLR2B) and Toll-like receptor 4 [TLR4]) were profiled along the broiler intestine. In addition, broiler intestinal segments were assayed for their total antioxidant capacity (TAC). Depending on PP inclusion level (i.e. 0, 750, 1,000 and 2,000 mg/kg diet) in the basal diets, 1-d-old Cobb broiler chickens (n = 500) were assigned into the following 4 treatments: CON, PP-750, PP-1000 and PP-2000. Each treatment had 5 replicates of 25 chickens with ad libitum access to feed and water. Data were analyzed by ANOVA and means compared using Tukey's honest significant difference (HSD) test. Polynomial contrasts tested the linear and quadratic effect of PP inclusion levels. Inclusion of PP increased (P ≤ 0.05) the expression of cytoprotective genes against oxidation, except CAT. In particular, the cytoprotective against oxidation genes were up-regulated primarily in the duodenum and the ceca and secondarily in the jejunum. Most of the genes were up-regulated in a quadratic manner with increasing PP inclusion level with the highest expression levels noted in treatments PP-750 and PP-1000 compared to CON. Similarly, intestinal TAC was higher in PP-1000 in the duodenum (P = 0.011) and the ceca (P = 0.050) compared to CON. Finally, increasing PP inclusion level resulted in linearly reduced (P ≤ 0.05) expression of NF-κB1, TLR4 and HSP70, the former in the duodenum and the latter 2 in the ceca. Overall, PP inclusion consistently up-regulated cytoprotective genes and down-regulated stress and inflammation related ones. The effect is dependent on PP inclusion level and the intestinal site. The potential of PP to beneficially prime bird cytoprotective responses merit further investigation under stress-challenge conditions.

The present study aimed to evaluate acid-insoluble ash (AIA) as an internal marker for the measurement the coefficient of total tract apparent digestibility (CTTAD) in rabbits through two experiments (E1 and E2). In E1, 48 rabbits were used to calculate the CTTAD of the same basal diet according to the European reference method (ERM), the AIA and the titanium dioxide (TiO2 with 1 g of TiO2/kg diet) techniques (n=16 rabbits/method). The effect of feed sample quantity on dietary AIA content was investigated and total collection of faeces was carried out to calculate marker recovery. In E2, 48 rabbits were allotted to three groups fed diets with no sugar beet pulp (SBP0) or with 100 (SBP100) and 200 (SBP200) g sugar beet pulp/kg (n=16 rabbits/group). Each group was divided into two subgroups, ERM and AIA (n=8 rabbits/subgroup), in which CTTAD was measured using the European reference and AIA method, respectively. In AIA subgroups, only 10% of the total daily faecal output was sampled from 9:00 to 9:30 am. Feed analysis in E1 showed that increasing sample quantity from 5 to 9 g did not affect the dietary AIA content; however, the analytical error was 7 and 5 times lower (P2/kg diet. Faecal marker recovery was 99.80±0.03 and 96.89±0.16% for AIA and TiO2, respectively. The CTTAD of dry matter (DM), did not differ between methods in E1, but a 5-fold higher variability (P2 technique in comparison with the ERM and AIA methods. Also, no differences in the CTTAD of DM between the ERM and AIA methods were found in E2. In conclusion, AIA is a reliable internal marker in rabbits and offers the possibility of measuring the CTTAD of diets with precision, when complete faecal collection or feed intake measurement is not possible.

Mycotoxins are a crucial problem for poultry production worldwide. Two of the most frequently found mycotoxins in feedstuffs are deoxynivalenol (DON) and fumonisins (FUM) which adversely affect gut health and poultry performance. The current knowledge on DON and FUM effects on broiler responses relevant for gut detoxification, antioxidant capacity, and health is still unclear. The aim of this study was to assess a range of selected molecular intestinal biomarkers for their responsiveness to the maximum allowable European Union dietary levels for DON (5 mg/kg) and FUM (20 mg/kg) in broilers. For the experimental purpose, a challenge diet was formulated, and biomarkers relevant for detoxification, antioxidant response, stress, inflammation, and integrity were profiled across the broiler intestine. The results reveal that DON significantly (p &lt, 0.05) induced aryl hydrocarbon receptor (AhR) and cytochrome P450 enzyme (CYP) expression mainly at the duodenum. Moreover, DON and FUM had specific significant (p &lt, 0.05) effects on the antioxidant response, stress, inflammation, and integrity depending on the intestinal segment. Consequently, broiler molecular responses to DON and FUM assessed via a powerful palette of biomarkers were shown to be mycotoxin and intestinal site specific. The study findings could be highly relevant for assessing various dietary bioactive components for protection against mycotoxins.

The present work was part of a project intended to evaluate whether organic selenium (Se) has the potential to protect against toxic effects exerted by cadmium (Cd). For this reason, 300 as-hatched, one-day-old broiler chickens were randomly allocated in four dietary treatments with five replicate pens per treatment. Chickens in T1 treatment, were offered a diet supplemented with 0.3 ppm Se (as Se-yeast), without added Cd, in T2 treatment, they were offered a diet with 0.3 ppm Se and 10 ppm Cd, in T3 treatment, they were offered a diet with 0.3 ppm Se and 100 ppm Cd, in T4 treatment, chickens were offered a diet supplemented with 3 ppm Se and 100 ppm Cd. Cadmium was added to the diets in T2, T3, and T4 as CdCl2. On the fourth and sixth weeks, liver and breast samples were obtained from two broilers per replicate pen. Relative gene expression levels of catalase (CAT), superoxide dismutase 1 (SOD1) and 2 (SOD2), methionine sulfoxide reductase A (MSRA) and B3 (MSRB3), iodothyronine deiodinase 1 (DIO1), 2 (DIO2), and 3 (DIO3), glutathione peroxidase 1 (GPX1) and 4 (GPX4), thioredoxin reductase 1 (TXNRD1) and 3 (TXNRD3), and metallothionein 3 (MT3) were analyzed by real-time quantitative PCR in liver, whereas the fatty-acid (FA) profile of breast muscle was determined by gas chromatography. Broilers supplemented with 0.3 ppm Se could tolerate low levels of Cd present in the diets, as there were no significant changes in the breast muscle FA profile, whereas excess Cd led to decreased polyunsaturated fatty acids (PUFAs), and in particular n-6 PUFA. Furthermore, treatments mainly affected the messenger RNA (mRNA) expression of SOD2, TXNRD3, and MT3, while age affected CAT, MSRB3, DIO2, DIO3, GPX4, TXNRD1, and MT3. In conclusion, dietary Se may help against the negative effects of Cd, but cannot be effective when Cd is present at excessive amounts in the diet.

The aim of this work was to assess the effect of dietary viable or heat inactivated probiotic forms (PF) combined or not with avilamycin (AV) used as a growth promoter, on broiler growth performance, nutrient digestibility, digestive enzyme activities, and expression of immune response related genes.Depending on the type of PF (i.e., no addition, viable, inactivated) and AV addition (no/yes), 450 one-day-old Cobb male broilers were allocated in the following 6 treatments according to a 3 × 2 factorial arrangement with 5 replicates of 15 broilers each for 6 wk: CoN: diet without any addition; CoN+A: combination of CoN with AV; ViP: viable PF - no AV; ViP+A: combination of ViP with AV; InP: inactivated PF - no AV; InP+A: combination of InP with AV.There were no interactions (P > 0.05) for overall performance parameters. In contrast, PF or AV addition improved BW gain (PPF= 0.015; PAV

The probiotic form (PF) type and its dietary administration in combination or not with avilamycin (AV) were investigated for their effects on broiler gut microbiota and expression of genes relevant for gut barrier and gut homeostasis. Depending on PF type (i.e. no addition, viable, inactivated) and AV addition (no/yes), 450 one-day-old Cobb male broilers were allocated in 6 treatments (CON, CON + A, ViP, ViP + A, InP and InP + A) according to a 3 × 2 factorial arrangement with 5 replicates of 15 broilers each for 42 days. Significant interactions (PPF × AV ≤ 0.05) between PF and AV administration were shown for the ileal mucosa-associated bacteria, the caecal digesta Lactobacillus spp., the molar ratio of the sum of valeric, hexanoic and heptanoic acids and the gene expressions of ileal and caecal IgA and ileal claudin 1. Avilamycin suppressed ileal digesta Lactobacillus spp. (PAV

A study was conducted to determine the effects of dietary crude glycerine (CG) on growth performance, gene expression of glycerine kinase (GK), nutrient digestibility, and litter quality in broiler chickens. A total of 400 one-day old broiler chickens were randomly assigned to 4 diets containing 0, 70, 140, or 210 g CG/kg with 5 replicate pens and 20 broiler chickens per pen. Diets were formulated by substituting CG for corn, on an equal apparent metabolizable energy (AMEn) basis, with similar crude protein and amino acid contents. Feed and water intake, weight gain, excreta moisture, litter score, and mortality were recorded weekly. The total tract apparent digestibility (TTAD) of nutrients and energy was determined at d 28, and GK expression was determined in liver homogenates at d 42. The overall feed intake and BW gain were affected quadratically ( P 0.001), with broiler chickens fed the diet with 70 g CG/kg diet showing the greater values, whereas feed conversion ratio and water intake increased linearly ( P 0.05 and 0.001, respectively) with increasing dietary CG. The linear increase ( P 0.01) in excreta moisture resulted in a linear reduction ( P 0.001) of litter quality with the increment of CG. At d 28, the TTAD of dry matter, organic matter, and nitrogen free extract were not affected; however, the dietary AMEn content tended to decrease ( P =0.07) when CG increased to 210 g/kg diet. Relative GK mRNA increased for CG inclusion up to 140 g/kg diet and reached a plateau thereafter, indicating a limiting capacity for dietary CG activation. In conclusion, dietary inclusion of CG up to 70 g/kg may have beneficial effects on growth performance of broiler chickens. Dietary inclusion up to 140 g CG/kg may be efficiently utilized, whereas greater inclusion (>140 g CG/kg) may impair growth of broiler chickens, reduce litter quality, and increase mortality.

Context Phytogenic applications in animal nutrition currently attract worldwide scientific attention for their potential to contribute positively to sustainable and high-quality animal production. However, further understanding and substantiation of dietary phytogenic functions is required. Aims The inclusion level of a phytogenic premix (PP) comprising functional flavouring substances from ginger, lemon balm, oregano and thyme was studied for its effects on broiler growth performance, carcass traits, nutrient digestibility, liver and meat total antioxidant capacity (TAC), and lipid oxidation. The expression of genes for nutrient transporter proteins (SGLT1, GLUT2, PEPT1, BOAT and LAT1), for FABP2 involved in cellular fatty acid uptake and metabolism, and for the mTORC1 complex relevant for protein synthesis were profiled along the intestine. Methods One-day-old Cobb broiler chickens (n = 500) were assigned to four treatments with five replicates of 25 chickens each. Starter (1–10 days), grower (11–22 days) and finisher (23–42 days) basal diets were supplemented with four levels of PP inclusion as treatments: 0, 750, 1000 and 2000 mg/kg diet, termed control, PP750, PP1000 and PP2000. Feed and water were available ad libitum. Data were analysed by ANOVA, taking the treatment as fixed effect. Statistically significant (P ≤ 0.05) effects were further analysed and means were compared using Tukey’s HSD test. Polynomial contrasts tested the linear and quadratic effect of PP inclusion levels. Key results Growth performance responses were not improved significantly (P > 0.05) by PP inclusion level. However, carcass (P = 0.030) and breast meat yield (P = 0.023) were higher in PP1000 than in the control. In addition, PP1000 had higher (P = 0.049) apparent metabolisable energy than PP2000 and the control. Increasing PP inclusion level increased breast (P = 0.005), thigh (P = 0.002) and liver (P = 0.040) TAC. Breast and thigh meat TAC reached a plateau at PP1000, whereas liver TAC continued to increase linearly. Lipid oxidation in breast meat and liver was delayed linearly (P ≤ 0.05) with increasing PP inclusion level. Expression of genes SGLT1, GLUT2, PEPT1, BOAT and FABP2 were not affected by PP inclusion. However, PP inclusion affected the expression of LAT1 (P < 0.001) in jejunum and of mTORC1 in duodenum (P = 0.010) and ceca (P = 0.025). In particular, expression increased with increasing PP inclusion level in a linear and quadratic pattern depending on the intestinal segment. Conclusions Overall, PP inclusion at 1000 mg/kg diet improved carcass and breast yield, dietary available energy, and overall meat and liver TAC. Preliminary evidence was highlighted for effects of PP in promoting expression of genes relevant for muscle protein synthesis. Implications This study has contributed new information on effects of a phytogenic premix on broiler meat yield and antioxidant capacity, digestibility, absorption and metabolic functions, further supporting phytogenic benefits for broiler production.

The effects of inclusion levels of a phytogenic feed additive (PFA), characterized by menthol anethol and eugenol, on broiler growth performance, nutrient digestibility, biochemical biomarkers and total antioxidant capacity (TAC) of plasma and meat, as well as on the relative expression of selected cytokines, were studied in a 42-d experiment. A total of 225 one-day-old male Cobb broiler chickens were assigned into 3 treatments, with 5 replicates of 15 chickens each. Chickens were fed maize-soybean meal basal diets following a 3 phase (i.e., starter, grower and finisher) feeding program. Depending on PFA inclusion level, treatments were: no PFA (PFA-0), PFA at 100 mg/kg (PFA-100) and PFA at 150 mg/kg (PFA-150). Feed and water were available ad libitum. Feed conversion ratio (FCR) during finisher phase was improved quadratically (P

The effects of substituting maize with crude glycerine (CG) as an energy source, on nutrient digestibility, liver glycerine kinase (GK) gene expression, and gut microbiota composition were investigated in 18 weaned pigs [30 days old, 7.8 kg initial body weight (BW)]. Pigs were fed one of three diets containing 0, 75 and 150 g CG/kg, formulated on equal digestible energy (DE) basis, with similar crude protein (210 g CP/kg dry matter; DM) and amino acid (12.9 g lysine and 8.2 g methionine+cystine/kg DM) content. The GK relative mRNA expression in liver and microbiota composition in ileal and caecal samples were assessed at 72 days of age. The digestibility of GE decreased (P Clostridium spp . and Lactobacillus spp . were linearly reduced (P

The aim of this study was to investigate the effect of dietary probiotic inclusion level on mucin composition (monosaccharide ratio), mucosal morphometry, mucus layer staining intensity, and mucus layer thickness along the broiler intestinal tract. One-day-old male Cobb broilers were administered maize-soybean meal basal (BD) diets for 42 d and depending on the feed additive used, broilers were allocated into the following 5 experimental treatments: control C (BD, no additive), treatment P1 (10(8) colony forming units of probiotic/kg of BD), treatment P2 (10(9) cfu of probiotic/kg of BD), treatment P3 (10(10) cfu of probiotic/kg of BD), and treatment A (2.5 mg avilamycin/kg of BD). Intestinal samples from duodenum, ileum, and cecum of 14- and 42-d-old broilers were collected and analyzed. Mannose (Man) decreased linearly with increasing probiotic level in duodenum (P=0.015) and ileum (P=0.042) of 14-d-old broilers. N-Acetyl-glucosamine and galactose decreased linearly (P=0.012 and P=0.001, respectively), while fucose increased linearly (P

Forty eight New Zealand × Californian rabbits were used to investigate the effects of diets with either a low (LDF) or high (HDF) inclusion of sugar beet pulp and wheat bran (feedstuffs rich in degradable fibre; DF) supplemented with or without soybean oil (20 g SO/kg) on liver weight and composition, according to a 2 × 2 factorial arrangement. At 77 days of age, the livers were excised, weighed, and analyzed for glycogen, cholesterol, and protein content and fatty acid (FA) composition. The HDF diets decreased liver relative weight (g/100 g body weight; P

The effect of a dietary phytogenic feed additive (PFA) inclusion level in mucin monosaccharide composition, mucosal morphometry and mucus histochemistry along the broiler intestinal tract was studied. Cobb male broilers (n = 525) were allocated into five experimental treatments that, depending on the type of addition in the basal diet (BD), were labeled as follows: C (BD based on maize–soybean meal with no other additions), E1 (80 mg PFA/kg BD), E2 (125 mg PFA/kg BD), E3 (250 mg PFA/kg of BD) and A (2.5 mg avilamycin/kg BD). Samples from duodenum, ileum and cecum of 14- and 42-day-old broilers were collected and analyzed. In 14-day-old broilers, treatments E2 and E3 had higher (P < 0.01) duodenal mannose than treatments C, E1 and A. Ileal mannose was lower (P < 0.05) in treatment C compared with PFA treatments, and ileal galactose (Gal) was higher (P < 0.01) in treatments E2 and E3 compared with C and A. Polynomial contrast analysis with respect to PFA inclusion level showed that in 14-day-old broilers there was a linear increase (P = 0.001) in duodenal mannose and a quadratic effect (P = 0.038) in duodenal N-acetyl-galactosamine with increasing PFA level. Ileal Gal and mannose increased linearly (P = 0.002 and P = 0.012, respectively) with PFA inclusion level. There were no significant differences between treatments in mucin monosaccharide molar ratios of 42-day-old broilers. However, increasing PFA inclusion level resulted in a linear decrease of ileal fucose (P = 0.021) and cecal N-acetylgalactosamine (P = 0.036). Experimental treatments did not differ (P > 0.05) regarding duodenal villus height (Vh), crypt depth (Cd) and Vh/Cd ratio, irrespective of broiler age and the intestinal segment examined. However, increasing dietary PFA inclusion level showed a pattern of linear increase of duodenal Vh/Cd ratio in 14-day-old broilers and ileal Vh in 42-day-old broilers (P = 0.039 and P = 0.039, respectively). Alcian Blue–Periodic Acid-Schiff (pH 2.5) staining of neutral and acidic mucins showed that the staining intensity of mucus layer in villi was fragment (i.e. tip, midsection and base) dependent, whereas in crypts it was dependent both on intestinal segment (i.e. duodenum, ileum and cecum) and fragment. Finally, mucus layer thickness did not differ (P > 0.05) between treatments, yet a pattern of linear increase (P < 0.05) with PFA inclusion level was observed in the duodenum of 42-day-old broilers. In conclusion, the dietary inclusion level of PFA modulated broiler intestinal mucin composition and morphology. Further studies are required to elucidate the physiological implications of such changes in host–microflora interactions.

The aim of this work was to evaluate the effect of three inclusion levels of a phytogenic feed additive (PFA) comprising a blend of essential oils from oregano, anise and citrus on broiler growth performance, nutrient digestibility and caecal microflora composition. Five hundred and twenty-five, 1-day-old, male Cobb broilers received a maize–soybean meal basal diet (BD) and depending on the type of addition were allocated in the following five experimental treatments for 6 weeks: BD–no additives (C), BD containing 80 mg PFA/kg diet (E1), BD containing 125 mg PFA/kg diet (E2), BD containing 250 mg PFA/kg diet (E3) and BD containing avilamycin at 2.5 mg/kg diet (A). Treatment A was used as a positive control due to the well-known function of avilamycin as an antimicrobial growth promoter in poultry. Overall broiler body weight gain (BWG) increased linearly (P=0.039) with increasing PFA level, however, the avilamycin treatment A did not differ (P≥0.05) from the PFA and C treatments. Overall feed intake (FI) decreased quadratically (P=0.046) with increasing PFA level, while the FI for treatment A did not differ from the rest of the treatments. The overall gain:feed ratio improved linearly (P n ) of experimental diets, respectively, with treatment E1 having the higher (P n (13.4 MJ/kg diet) and treatment A being not different from the four other treatments. On the other hand, the avilamycin treatment A had a significantly (P Lactobacillus (P=0.002), Bifidobacterium (P=0.001) and Gram positive cocci (P=0.007) concentration with increasing PFA level at 42 d old broilers. In addition, caecal coliforms at 14 d old broilers were significantly (P In conclusion, this small scale study provides additional evidence that phytogenic efficacy in broilers depends on the feed inclusion level used and the broiler growth period with most of the PFA overall beneficial effects seen mainly in the finisher growth period. Further work is warranted in order to evaluate phytogenic efficacy with different basal diets and under commercial production conditions.

Summary A total of 128 broilers were used to investigate the effect of selenium (Se) on fatty acid (FA) composition and oxidative stability of lipids in the breast muscle tissue. There were 4 replicates of 4 dietary treatments: T1 (basal diet with no added Se), T2 (T1 with 0.15 mg Se added per kg diet), T3 (T1 with 0.3 mg Se added per kg diet) and T4 (T1 with 3.0 mg Se added per kg diet). A yeast source was used for added Se. Breast muscle tissue was collected from two chickens per replicate pen for the determination of Se concentration by ICP-MS, FA profile by GC and lipid oxidation using thiobarbituric acid reactive substances method. Addition of supranutritional Se levels to chicken diets leads to the production of Se-enriched meat. Consumption of 100 g of breast meat from chickens fed diets supplemented with 0.15, 0.3 and 3 mg Se per kg of diet can provide 26, 41 and 220 μg of Se, respectively. Long-chain polyunsaturated fatty acids namely C20:3n-6, C20:4n-6, C20:5n-3, C22:5n-3 and C22:6n-3 increased linearly (p = 0.047, p

Summary The study aimed to describe the interactions between nutrition and caecal bacteria abundance using odd-numbered and branched-chain fatty acids (OBCFA). Following a 2 × 2 factorial design, 47 rabbits were fed diets with two levels of digestible fibre (DF, 180 vs. 260 g/kg DM) supplemented with soybean oil (SO, 20 g/kg) or not. At 77 days of age, fatty acid (FA) composition was determined in caecal contents. The microbial origin of OBCFA vs. dietary FA in caecal contents was illustrated by clustering of these FA in the loading plots of principal component analysis. The contribution of odd-numbered FA in the OBCFA pattern was increased (p

The study aimed to characterize shifts in caecotroph microbial FA induced by different dietary digestible fibre (DF) and soybean oil (SO) contents, in fattening rabbits. Following a 2 × 2 design, diets with either a low (LDF) or high (HDF) digestible fibre content (180 and 260 g DF/kg DM, respectively) supplemented with or without 20 g of SO/kg, were fed to 12 rabbits (n = 48 in total) for a period of 77 days. At an age of 63 days, a light plastic flat collar was applied on animals for 24 h to prevent caecotrophy. Gas chromatography was used to determine the odd-numbered (ONFA), branched-chain (BCFA), hydroxy (OHFA) and other FA in freeze-dried caecotroph samples. A high dietary DF level increased ONFA (P

The aim of this work was to investigate the effect of inclusion levels of a 5-bacterial species probiotic in broiler nutrition. Five hundred twenty-five 1-d-old male Cobb broilers were allocated in 5 experimental treatments for 6 wk. The experimental treatments received a corn-soybean coccidiostat-free basal diet and depending on the addition were labeled as follows: no addition (C), 10(8) cfu probiotic/kg of diet (P1), 10(9) cfu probiotic/kg of diet (P2), 10(10) cfu probiotic/kg of diet (P3), and 2.5 mg of avilamycin/kg of diet (A). Each treatment had 3 replicates of 35 broilers each. Treatment effects on broiler growth performance and biomarkers such as ileal and total tract nutrient digestibility, plasma Ig concentration, and cecal microflora composition were determined. Differences among treatments were considered significant when P < or = 0.05. Overall BW gain was significantly higher in treatment P1 (2,293 g) compared with P2 (2,163 g), C (2,165 g), and P3 (2,167 g), with A (2,230 g) being intermediate and not different from P1. Overall feed conversion ratio values were similar and significantly better for P1 (1.80) and A (1.80) compared with P2 (1.87), C (1.89), and P3 (1.92). Ileal apparent digestibility coefficients (ADC) of CP and ether extract were higher in A. Generally, treatments A and P1 showed an improved total tract ADC for DM, organic matter, ash, ether extract, and AME(n) values. The total tract ADC of CP was higher in P1, C, and P2. There were no differences between treatments regarding plasma Ig in 14- and 42-d-old broilers. Treatments P2 and P3 were effective at beneficially modulating cecal microflora composition. In particular, the lower cecal coliform concentration (log cfu/g of wet digesta) was seen in P2 (6.12) and P3 (4.90) in 14- and 42-d-old broilers, respectively, whereas at 42 d, P3 and P2 had the highest Bifidobacterium (8.31; 8.08) and Lactobacillus concentrations (8.20; 7.86), respectively. It is concluded that probiotic inclusion level had a significant effect on broiler growth responses, nutrient ADC, AME(n), and cecal microflora composition.

Summary Background & aims Little is known about fasting effects on gut bacterial metabolism. As probiotics are purported to be beneficial for health, this study aimed to investigate the response of gut microbial metabolism on fasting with or without probiotic administration. Methods Sixty male adult Wistar rats were allocated to six experimental treatments, for 6 days, arranged under three nutritional schemes namely: (a) ad libitum feeding (control), (b) fasting for 3 days and re-feeding for the remainder (re-fed) and (c) fasting for 6 days combined with parenteral liquid treatment during the last 3 days (starved). Each nutritional scheme had one non-probiotic and one probiotic treatment receiving orally Lactobacillus acidophilus . Rat caecal digesta were analyzed for bacterial enzyme activities and volatile fatty acids (VFA). Results Fasted rats had significantly lower activities of α-galactosidase, α-glucosidase and β-glucosidase and higher activities of β-galactosidase and azoreductase compared to control and re-fed rats, irrespective of probiotic administration. Results were variable regarding cholylglycine hydrolase (CGH), while there were no differences between treatments regarding β-glucuronidase and arylsulfatase activity. Fasted rats had significantly lower caecal VFA concentration and different fermentation patterns. L. acidophilus resulted in significantly reduced azoreductase activity and increased caecal acetate levels in fasted rats. Re-feeding appeared to restore most enzyme activities, fermentation intensity and to some extent fermentation patterns at control treatment levels. L. acidophilus resulted in significantly reduced CGH activity and increased butyrate levels in re-fed rats. Conclusion The results indicate a health beneficial potential of L. acidophilus in fasted and re-fed nutritional states via reduction of harmful azoreductase and CGH activities and promotion of useful VFA components for colonic function and health.

The dietary supplementation of Saccharomyces cerevisiae var. boulardii was evaluated in broilers challenged or not challenged with Salmonella Enteritidis (SE) using a 2 × 2 factorial arrangement. Depending on yeast inclusion at 0 (C) or 1 × 10⁹ cfu/kg diet (Y) and SE challenge (0 or log 6.3 cfu/bird) on d 15, the experiment had four treatments C, Y, C-SE, and Y-SE, respectively. Each treatment had seven replicate floor pens with 15 broilers. Growth performance responses were determined weekly and overall for the 5 week experimental period. Salmonella levels and prevalence in ceca, cloacae, and carcass skin were determined by culture procedures, while cecal microbiota was determined by real time PCR. Yeast supplementation had no effect (PY0.05) on growth performance. For the overall post SE-challenge period (i.e., wk 3 to wk 5), Salmonella reduced body weight gain (BWG) (PSE0.001), feed intake (FI) (PSE = 0.032), and the European production efficiency (EPEF) factor (PSE = 0.005). Broilers Y-SE had higher (P0.001) overall BW gain compared to C-SE ones. Overall mortality was 2.14% and did not differ (P0.05) between treatments. Reduced Salmonella levels in the cloacae (P = 0.014) and on the breast skin (P = 0.006) and lower prevalence on the neck skin (P = 0.007) were noted for treatment Y-SE compared to C-SE. Yeast supplementation did not have an effect (P0.05) on cecal microbiota composition at d 1 and d 21 post SE-challenge. On the contrary, SE-challenge reduced cecal levels of total bacteria (PSE = 0.002), E. coli (PSE = 0.006), Bifidobacterium spp. (PSE = 0.006), Bacteroides spp. (PSE = 0.010), and Clostridial populations belonging to cluster I and cluster XIVa, (PSE = 0.047 and PSE = 0.001, respectively) on d 1 post SE-challenge. At 21 d post SE-challenge, only the levels of cecal Lactobacillus spp. (PSE = 0.001) and Bifidobacterium spp. (PSE = 0.049) were reduced compared to the non SE-challenged groups. In conclusion, yeast supplementation in SE challenged broilers (Y-SE) was beneficial for growth performance and reduced Salmonella presence compared to C-SE ones. The disturbance of cecal microbiota balance by SE merits further investigation for potential implications in gut and overall bird health.

The effect of oligofructose (FOS) or trans-galactooligosaccharides (TOS) inclusion at 10 g/kg diet for growing pigs, at the expense of cornstarch (CON), on growth performance, nutrient digestibilities, and faecal physicochemical parameters was determined. In addition, concentration of volatile fatty acids (VFA) and bacterial α-glucosidase, β-glucosidase, β-galactosidase and β-glucuronidase activities in faeces were determined as markers of intestinal microflora metabolic activity. Twelve 35-day-old castrated pigs were allocated to CON, FOS and TOS dietary treatments for an experimental period of 4 weeks. Dietary treatments did not affect animal growth performance. FOS and TOS pigs tended to have a lower faecal output than CON pigs by approximately 6.0% and 1.7% respectively. FOS and TOS did not affect total tract apparent digestibility of dry matter, organic matter, fat, crude protein, crude fiber, ADF and cellulose. Digestibility of carbohydrate components such as nitrogen free extract and NDF was significantly (P ≤ 0.05) higher in TOS pigs compared to CON and FOS. Digestibility of hemicellulose in TOS pigs did not differ from FOS but was significantly higher from CON. The experimental period had a significant effect on most of the parameters determined. There were no significant (P ≤ 0.05) differences between treatments regarding total VFA concentration and faecal bacterial enzyme activities. Nevertheless, an increased saccharolytic activity in the gut of FOS and TOS pigs could be postulated by the trends for apparently higher VFA concentration and bacterial α-glucosidase and β-galactosidase activities.

(1) The objective of this study was to determine whether the dietary inclusion of Trichosporon mycotoxinivorans (TRM) could suppress the detrimental effects of ochratoxin A (OTA) on the immune system of broiler chicks. (2) Six experimental treatments were tested in 300 1-d-old broiler chicks. Treatments included addition to a standard broiler ration of neither OTA nor TRM (Diet 1), OTA alone (500 microg/kg), OTA plus TRM at three inclusion rates (10(4) CFU/g of feed, 10(5) CFU/g, 10(6) CFU/g) and TRM alone at 10(5) CFU/g of feed. The ration was fed to chicks for 42 d. (3) Blood samples were collected at d 10, 20, 30 and 40 and macrophages and heterophils were isolated. The following variables were determined in macrophages and heterophils activated by phorbol myristate acetate (65 microM): cell viability, total cell-associated urokinase-plasminogen activator (u-PA), membrane-bound u-PA, free u-PA binding sites and superoxide production. (4) There was a decrease in the viability of macrophages and heterophils from chicks receiving OTA-contaminated feed compared to the viability of cells from control birds at d 40. Dietary TRM completely blocked the effect of OTA on cell viability; all three inclusion rates were equally effective. There was a decrease in total cell-associated and membrane-bound u-PA in macrophages and heterophils of chicks receiving OTA-contaminated feed compared to the corresponding values in control birds for heterophils at d 30 and 40 and for the macrophages at d 40. (5) Similarly, dietary TRM abolished the effect of OTA on total cell-associated and membrane-bound u-PA activity. All three inclusion rates of yeast were equally effective. Heterophils, but not macrophages, isolated from chicks receiving OTA-contaminated diet produced less superoxide anion compared to all other diet groups at d 30 and 40. (6) The immune system is a primary target of OTA toxicity in broilers: several functional properties of macrophages and heterophils were depressed in chicks fed OTA-contaminated feed. There was a delay of 30d before the immunosuppressive effect became apparent. The dietary inclusion of TRM completely blocked the detrimental effects of OTA on several immune properties in broilers.

Sixteen piglets were used to determine the effect of vitamin E supplementation on several functional properties of macrophages and neutrophils obtained from weaned piglets. Piglets, immediately following weaning, were assigned to one of three experimental groups: control (no vitamin E supplementation), low level of vitamin E supplementation (100 mg DL-alpha-tocopheryl acetate/kg diet) and high level of vitamin E supplementation (300 mg DL-alpha-tocopheryl acetate/kg diet). Supplementation of vitamin E lasted for a period of 36 days, following a 3-day adaptation period after weaning. Blood samples were collected on days 0, 12, 24 and 36 of the experimental period, monocytes/macrophages and neutrophils were isolated and the following parameters were determined in macrophages and neutrophils activated by phorbol myristate acetate: total cell-associated and membrane-bound urokinase plasminogen activator (u-PA) activity and superoxide anion production. Results showed that macrophages and neutrophils isolated from piglets that received supplemental vitamin E had higher (P0.05) total and membrane-bound u-PA activities as well as higher (P0.05) superoxide anion production compared with the values of the corresponding cells obtained from control piglets on day 12 of the experimental period. Both levels of vitamin E supplementation (low and high) were equally effective. In contrast, vitamin E supplementation had no effect (P0.05) on total and membrane-bound u-PA activities and superoxide anion production by porcine macrophages and neutrophils on days 24 and 36 of the experimental period. In conclusion, the low level of vitamin E supplementation is recommended for piglets for the first 2 weeks after weaning.

Near infrared spectroscopy (NIRS) is widely used in the flour milling industry for rapid determination of moisture and protein in wheat. However, these measurements give little indication of the nutritive value of wheat when fed to poultry or pigs. Accurate estimates of nutritive value require specialist facilities and are time-consuming and costly. Accordingly, prediction from chemical or NIRS measurements would be of some considerable benefit. In the current study 160 samples of wheat, representing 24 different varieties, were used to generate NIRS calibration equations for chemical, nutritive and agronomic characteristics. Predictions of chemical constituents in wheat were very accurate. Coefficients of determination (r2) were 0·94 for dry matter, 0·90 for crude protein, 0·97 for ash, 0·78 for starch and 0·98 for oil. True metabolizable energy in broiler chickens was predicted more accurately (r2 = 0·52 for adult birds, 0·74 for young birds) than apparent metabolizable energy (r2 = 0·45). Digestible energy (r2 = 0·17) and nitrogen digestibility (r = 0·22) in pigs were not predicted very accurately on a smaller subset (n = 33). Agronomic characteristics were predicted very accurately (r2 = 0·98 hardness, 0·80 bushel weight, 0·99 thousand-grain weight). Predictions of nutritive value of wheat from chemical or agronomic characteristics are very inaccurate, since coefficients of determination vary from zero to 0·25. It is concluded that NIRS can accurately estimate the chemical composition of wheat, but accurate prediction of nutritive value is reduced by animal variation. Nevertheless, NIRS is potentially more reliable for assessing nutritive value than chemical composition or agronomic characteristics.

Digestion and performance of lactating dairy ewes fed diets with soy hulls (SH), with or without dietary fat supplements, as a replacement for maize in concentrate diets were evaluated in digestion and milk performance trials. In the digestion trial, three fistulated wether sheep were used in a 3×3 Latin square experimental design. The diets consisted of 400 g hay+600 g concentrate with 60% maize (control treatment), or 60% soy hulls (SH treatment), or 60% soy hulls+5% soybean oil (SHF treatment). Diets' DM and OM digestibilities were similar among treatments, while the replacement of maize by SH caused a significant reduction on CP, EE, and NFE digestibilities, and a significant increase on NDF and ADF digestibilities. The addition of 5% soybean oil to SH concentrate had similar effect on nutrients digestibility, as SH. Concentrates DM and CP degradabilities, pH values and VFA concentrations in rumen liquor were also similar among treatments. In the performance trial, 45 lactating ewes were used, which were fed daily, for 7 weeks, 800 g hay+1200 g concentrates of the type as in the digestibility trial. Milk yield and FCM yield increased by 3 and 33% ( p p p p −1 ) was higher for the SHF diet, followed by SH and then by the control diet. All ewes gained weight during the experimental period, but the BW change was not significant among treatments. The results showed that SH can replace cereal grains in lactating ewe diets, and minimize the milk fat-depressing effects of grain-based diets.

The objective of this study was to describe and evaluate the feeding system of milking sheep in a typical mountainous area (Feneos of Korinthos, Greece), which comprises eight village communities. In this area, 23 000 sheep and 10000 goats are kept under an extensive livestock farming system and a semi-intensive feeding system. Available pasture, cultivated land and meadows comprise 16 800 ha, 4540 ha and 443 ha, respectively. Sheep graze throughout the year but herbage intake is sufficient to meet the animals' requirements only from April to June. During the summer, sheep graze cereal stubble and other crop residues left after harvesting. From October to March, animals are provided with concentrates and roughages. All of the roughages and part of the concentrates are farm grown. On an annual basis, concentrates, roughages and ingested herbage from grazing contribute 36%, 26% and 38% of total energy requirements, respectively, whereas supplementary feed only exceeds the animals' CP requirements. During winter the nutrient requirements of the sheep are not met, consequently animals lose weight (BC score 1.5 – 2.5), but this is restored in the spring when there is usually plenty of grass. The rations fed during winter are not properly balanced, and there is usually an energy deficit and a protein surplus. No mineral supplements are fed, and some (about 18%) sheep suffer from subclinical Zn and Se deficiencies. It is concluded that the genetic potential of the sheep, under the feeding system of a typical mountain management region, has not been reached. Costs and benefits of supplementation need careful evaluation.

The effect of dietary acidifier (AC) supplementation in combination or not with avilamycin (AV) was evaluated on broiler performance, nutrient digestibility, gizzard pH, activity of digestive enzymes and relative expression of cytokines. A 2 × 2 factorial experimental design with inclusion of AC (0 and 1 g/kg of diet) and AV (0 and 2.5 mg/kg of diet) as the main factors was implemented. Subsequently, 544-day-old male Cobb broilers were allocated in four treatments (i.e. C – no additions, AC; AV and ACAV), each having eight replicates of 17 broilers, for 6 weeks. Overall bodyweight gain (BWG) was improved (PAC = 0.035) by AC addition. BWG and feed conversion ratio were improved (P ≤ 0.05) by AV addition in all growth periods and overall. Dry matter, organic matter digestibility and energy retention were improved by AC (PAC = 0.050, PAC = 0.045 and PAC = 0.020, respectively), or AV (PAV = 0.018, PAV = 0.013 and PAV = 0.012, respectively). In addition AV supplementation improved digestibility of ether extracts (PAV = 0.022) and crude protein (PAV = 0.042). Pancreatic trypsin activity was lower in AC (PAC = 0.027) or AV (PAV ≤ 0.001) supplemented diets. An AC × AV effect on gizzard pH (PAC × AV = 0.034) was noted. Expression of interferon-γ (PAV = 0.010), interleukin (IL) 18 (PAV = 0.048) and inducible nitric oxide synthase 2 (PAV = 0.035) in spleen were downregulated by AV. An upregulation of IL10 (PAC × AV = 0.018), IL18 (PAC × AV = 0.002) and IL22 (PAC × AV = 0.030) in the caecal tonsils was shown for the combined ACAV supplementation. In conclusion, dietary AC supplementation improved performance, nutrient digestibility and energy retention, irrespective of AV inclusion. This study has provided evidence for potential links between digestive and immune function indices with growth performance improvements.

The purpose of this study was to investigate the preservation time of fresh olive tree leaves (OTL) after treatment with ammonia (AM-OTL) solution, the improvement of its nutritive value and the effect on milk yield and milk composition of dairy ewes. The crude protein (CP) content of OTL treated with a 25% aqueous ammonia solution (5 1 per 100 kg fresh OTL) increased from 96 to 180 g kg−1 dry matter (DM) just after the bags were opened, and 164 and 155 g kg−1 DM 2 and 12 months after treatment, respectively. The non-protein nitrogen of AM-OTL was 312 g kg−1 CP compared with 100 g kg−1 CP of fresh untreated OTL (F-OTL). Results of a digestibility experiment showed that the apparent digestibility (g kg−1) of AM-OTL increased (P < 0.05) for CP, crude fibre (CF), neutral detergent fibre and acid detergent fibre by 47.4%, 19.4%, 13.3% and 10.5% and decreased for ether extractives (EE) by 10.7% in relation to F-OTL. The nutritive value was estimated to be 4.60 and 4.81 MJ of net energy of lactation (NEL) kg−1 DM for F-OTL and AM-OTL, respectively. In the second experiment 24 dairy lactating ewes of the Karagouniko breed (55 kg of average BW) were used and divided into two equal groups, immediately after weaning of their lambs. Using a cross-over design, ewes in Group 1 were fed a diet (A) with 1.2 kg AM-OTL + 0.25 kg wheat straw+ 0.88 kg concentrates. Ewes in Group 2 were fed a diet (B) with 0.8 kg alfalfa hay +1 kg concentrates. Diet (A) caused an increase (P < 0.05) in soluble CP and a reduction (P < 0.05) in casein content of milk. Fatty acid composition of milk fat was affected by AM-OTL with a reduction (P < 0.001) of short chain (C6 to C16:l) and an increase of long chain (C18, C18:1) fatty acids. The results showed that ammonia treatment preserves OTL, improves their nutritive value and allows their inclusion in ruminant diets by replacing other expensive roughages.

1. Probiotics are beneficial microbes that are currently considered as alternatives to antimicrobial growth promoters (AGP) in animal nutrition. In addition, they are purported to suppress pathogens in the gut via the concept of competitive exclusion (CE). The aim of this work was to evaluate the efficacy of a multistrain probiotic against Salmonella enteritidis (SE) in broilers. 2. Three hundred and four, one-day-old, male Cobb broilers were fed a corn-soybean basal diet and were allocated to four experimental treatments for 6 weeks. Three of the four treatments were challenged with SE. Treatments were: C- (Negative Control, no SE challenge), C+ (Positive control, SE challenge, no other additives), AB (SE challenge + Avilamycin used as AGP) and PFW (SE challenge + probiotic in feed and water). Morbidity, mortality, incidence of Salmonella positive broilers, cecal microflora composition and antibody response (IgA and IgG) in the blood and intestine were determined as biomarkers of probiotic CE efficacy against SE. 3. The three SE challenged treatments had a significantly higher morbidity compared to C- during the first 2 weeks of age, while there were no differences between treatments regarding mortality. Salmonella shedding was evidenced in 75 and 50% of the birds at 5 and 21 d post-challenge, respectively. Microbiological analysis of cecal digesta in 42-d-old broilers revealed that while broilers in treatment C+ were 100% Salmonella positive, the broilers in treatments PFW and AB were 50% positive and in addition they had lower Salmonella levels (CFU/g digesta) by 27 logs compared to C+. At the age of 42 d significantly higher IgA and IgG specific immune responses against SE were detected at systemic and at intestinal level only for the positive control treatment (C+). 4. In conclusion, the lack of significant levels of specific IgA and IgG against SE at systemic and intestinal level combined with the lower prevalence of SE positive broilers and the lower cecal SE levels in treatments AB and PFW compared to C+, suggest that treatments PFW and AB were efficacious at reducing SE.

Fifteen piglets were used to determine the effect of vitamin E supplementation on the number of CD4-immunoreactive (CD4+) T-lymphocytes, CD8-immunoreactive (CD8+) T-lymphocytes and IgA-immunoreactive (IgA+) B-lymphocytes per follicle in the Peyer's patch of distal ileum and the mesenteric lymph nodes of weaned piglets. Piglets, following a 3-day adaptation period after weaning, were assigned to one of three experimental groups: control (no vitamin E supplementation), vitamin E supplementation of 100 mg/kg of diet and vitamin E supplementation of 300 mg/kg of diet. Supplementation of vitamin E lasted for a period of 36 days. The basal diet contained 80 mg alpha-tocopherol/kg of diet. All piglets were killed at day 39 after weaning and samples of the distal ileum and adjacent mesenteric lymph nodes were collected. The number of cells for each lymphocyte subset was counted in the Peyer's patch and the mesenteric lymph nodes follicles, in cryostat sections processed for immunohistochemistry. Results showed that vitamin E supplementation (300 mg/kg diet) of piglets caused an increase (P0.05) in the number of IgA+ B-lymphocytes in the Peyer's patch, but not in the mesenteric lymph nodes, compared with the corresponding values in control animals. Vitamin E supplementation had no effect (P0.05) on the number of CD4+ and CD8+ T-lymphocytes in the follicles of the Peyer's patch and the adjacent mesenteric lymph nodes. Thus, vitamin E had relatively minor effects on distribution of the major immunocompetent cells in the gut. The numbers of CD4+ and CD8+ T-lymphocytes as well as IgA+ B-lymphocytes per follicle were higher by 26-77% (P0.05) in the mesenteric lymph nodes than the corresponding values in the Peyer's patch.

It is evident that quantitative information on different microbial groups and their contribution in terms of activity in the gastrointestinal (GI) tract of humans and animals is required in order to formulate functional diets targeting improved gut function and host health. In this work, quantitative information on levels and spatial distributions of Bacteroides spp, Eubacterium spp, Clostridium spp, Escherichia coli, Bifidobacterium spp and Lactobacillus/Enterococcus spp. along the porcine large intestine was investigated using 16S rRNA targeted probes and fluorescent in situ hybridisation (FISH). Caecum, ascending colon (AC) and rectum luminal digesta from three groups of individually housed growing pigs fed either a corn-soybean basal diet (CON diet) or a prebiotic diet containing 10 g/kg oligofructose (FOS diet) or trans-galactooligosaccharides (TOS diet) at the expense of cornstarch were analysed. DAPI staining was used to enumerate total number of cells in the samples. Populations of total cells, Bacteroides, Eubacterium, Clostridium and Bifidobacterium declined significantly (P < 0.05) from caecum to rectum, and were not affected by dietary treatments. Populations of Lactobacillus/Enterococcus and E. coli did not differ throughout the large intestine. The relative percent (%) contribution of each bacterial group to the total cell count did not differ between caecum and rectum, with the exception of Eubacterium that was higher in the AC digesta. FISH analysis showed that the sum of all bacterial groups made up a small percentage of the total cells, which was 12.4%, 21.8% and 10.3% in caecum, AC and rectum, respectively. This supports the view that in swine, the diversity of GI microflora might be higher compared to other species. In terms of microflora metabolic activity, the substantially higher numerical trends seen in FOS and TOS treatments regarding total volatile fatty acid, acetate concentrations and glycolytic activities, it could be postulated that FOS and TOS promoted saccharolytic activities in the porcine colon.

Three dietary combinations of probiotics and phytogenics administered sequentially or continuously were assessed for their effects on broiler performance, nutrient digestibility, caecal microbiota composition, volatile fatty acid (VFA) pattern and plasma total antioxidant capacity (TAC). One-day-old Cobb male broilers (n = 525) were allocated to five experimental treatments for 42 days. Each treatment had three replicates of 35 broilers each. Depending on the time-frame of sequential or continuous addition in the basal diet (BD) of the probiotics (108 CFU/kg BD) and phytogenics (125 mg/kg BD), experimental treatments were: control (no additions, Days 1–42); PE1, probiotic Days 1–14 and phytogenic Days 15–42; PE2, probiotic Days 1–28 and phytogenic Days 29–42; PE3, probiotic and phytogenic continuously (Days 1–42); and A (2.5 mg avilamycin/kg diet, Days 1–42). There was no loss of probiotic viability upon mixing with the phytogenic for up to 7 days. Overall bodyweight gain was higher (P = 0.025) in the control and PE1 than PE2 and PE3, whereas Treatment A was intermediate and not different from the rest. Overall feed intake in the control was the highest (P = 0.003). Treatments PE1, PE2 and PE3 had better (P = 0.004) overall feed conversion ratio (FCR) than the control. The FCR in PE1 and PE3 did not differ from A, which had the best FCR. Mortality did not differ among treatments. Nutrient digestibility and caecal microbiota composition did not differ among treatments. However, differences (P ≤ 0.05) in caecal VFA pattern were noted, with the control having the lowest acetate molar ratio as well as the highest sum of valeric, hexanoic and heptanoic acids. Plasma TAC in the control was lower than in PE3, PE1 and PE2 at Day 14 (P < 0.001) and PE1 (P = 0.003) at Day 42. The concept of sequential or throughout probiotic and phytogenic administration showed promising results regarding feed efficiency and plasma TAC.

The effect of the dietary inclusion level of a three-species probiotic on broiler performance, nutrient digestibility, caecal microbiota composition and volatile fatty acid (VFA) pattern was evaluated. Day-old Cobb broilers (n = 448) were allocated in four treatments for 6 weeks. Each treatment had four replicates (two per gender) of 28 broilers each. Depending on the type of addition per kg basal diet, treatments were C (no other addition), PL (108 colony forming units of probiotic), PH (109 colony forming units of probiotic) and A (2.5 mg avilamycin). Overall bodyweight gain was better (P = 0.002) in PL and PH than in the control (2082 g) by 8.7% and 7.5%, respectively, while treatment PL did not differ from A (2341 g), which showed the highest bodyweight gain. The ileal and total-tract apparent digestibility of DM and the apparent metabolisable energy content corrected for N improved linearly (P ≤ 0.05) with the probiotic level. Fluorescent in situ hybridisation analysis showed caecal Bifidobacterium levels to increase linearly (P = 0.006) with the probiotic level. Probiotic administration resulted in altered caecal VFA patterns compared with the control. Gender effects (P ≤ 0.05) were noted for caecal levels of C. histolyticum group, Bacteroides fragilis group and Streptococcus spp., while interactions (P ≤ 0.05) of treatment with gender were seen for Bifidobacterium and all VFA components, except for acetate. In conclusion, beneficial effects on bodyweight gain, DM digestibility, apparent metabolisable energy content corrected for N, caecal Bifidobacterium levels and VFA patterns were noted with both probiotic inclusion levels.

One hundred and forty five pigs, initially weighing 28 kg, were used in an experiment to evaluate the effect of partially replacing soya bean meal (SBM) with cottonseed meal (CSM) on the performance of fattening pigs. The experiment of 13 weeks was divided into two periods, prefattening (28–60 kg) and final fattening (60 kg until slaughter). For each period two diets with SBM and CSM were formulated to have the same contents of crude protein, lysine and sulphur amino acids. Treatment I consisted of SBM, while in treatment II, CSM and SBM (3:1) were used and lysine-HCl was added. Energy and nutrient digestibility were also investigated in each fattening period. Feed intake was unaffected by treatment. During the prefattening period weight gain was depressed 11% on Treatment II; this depressive effect remained for the females only during the finishing phase. The feed-conversion ratio was consequently increased with this treatment, on average by 13.5% and 6.6% for prefattening and final fattening phase respectively. Apparent digestibility of gross energy and crude fibre was significantly lower with the CSM-than with the SBM-containing diet. Also, the CSM-containing diet had lower digestibility coeffients for threonine, isoleucine, valine, aspartic acid and serine. The carcasses of the pigs given CSM had a significantly ( P ⩽ 0.001) lower mean percentage value in meat cuts.

Two experiments were conducted to evaluate the effects of sugar beet molasses (0, 8 and 16%) with or without lasalocid sodium (33 ppm) on rumen liquor and blood parameters of sheep, and on fattening reared Karagouniko breed lambs. In Expt. 1 six adult Karagouniko fistulated wethers were fed six different pelleted diets. The results showed that lasalocid decreased (P less than 0.01) total VFAs concentration in the rumen (7.1%) by reducing (P less than 0.001) acetic, n-butyric and iso-valeric acids while increased (P less than 0.001) propionic acid concentrations. The same trends were observed in molar proportions of the individual VFAs. The total VFAs production was also depressed by 13.75% when molasses were used at the level of 16%. No interaction among the main treatments was observed with the exception of L X M with 16% molasses where the propionic acid concentration decreased, and those of butyric and iso-valeric acids increased. Lasalocid did not alter the mean values of rumen liquor pH and ammonia nitrogen concentration while molasses increased the later parameter. No significant differences on dietary protein degradability were observed among treatments. Lasalocid reduced the beta-HBA concentration by 13% in the blood while molasses increased it (P less than 0.001). Free fatty acids' concentration was decreased by the molasses inclusion to the diets, while lasalocid didn't alter blood glucose concentration. Finally, the number of protozoa in the rumen liquor was reduced by 49 and 70% at 0 and 5 hours post feeding respectively with the addition of lasalocid to the diets, regardless of the use of molasses. In Expt. 2 utilized 86 weaned male Karagouniko fattening lambs, with initial mean LW 13 kg, for 60 days. They were assigned to six groups and fed ad libitum with diets of the same composition as in Exp 1. Lasalocid increased the growth rate by 8.6%, reduced feed consumption by 4.8% (P less than 0.005) and improved feed conversion by 11.8%. Molasses also improved growth rate by 16-34% and increased feed consumption by 15-22%. The interaction of L X M to feed consumption was significant (P less than 0.001). None of the treatments had an effect on carcass characteristics. Finally lasalocid showed its positive action when it was used in combination with molasses in ruminants' diets.

The aim of this work was to investigate the efficacy of a new multibacterial species probiotic in broiler nutrition. The probiotic contained 2 Lactobacillus strains, 1 Bifidobacterium strain, 1 Enterococcus strain, and 1 Pediococcus strain. Four hundred 1-d-old male Cobb broilers were allocated in 4 experimental treatments for 6 wk. The experimental treatments received a corn-soybean basal diet and were as follows: "control," with no other additions; "probiotic in feed and water," (PFW) with probiotic administered at 1 g/kg of feed for the whole period and in water on scheduled intervals during the first 4 wk; "probiotic in feed," (PF) with probiotic in feed as in PFW; and "antibiotic," (AB) with addition of avilamycin at 2.5 mg/kg of feed. Salinomycin Na was used as a coccidiostat. Each treatment had 5 replicates of 20 broilers. Treatment effects on parameters of broiler performance and cecal microbial ecology were determined. Broiler BW, feed intake, and feed conversion ratio were determined on a weekly and overall basis. Cecal microflora composition, concentration of volatile fatty acids, and activities of 5 bacterial glycolytic enzymes (alpha-galactosidase, beta-galactosidase, alpha-glucosidase, beta-glucosidase, and beta-glucuronidase) were determined at the end of the experiment. Overall, treatment PFW displayed a growth-promoting effect that did not differ from AB. Overall, feed conversion ratio in treatment AB was significantly better (P < or = 0.01) than the control treatment, whereas treatments PFW and PF were intermediate and not different from AB. Concentrations of bacteria belonging to Bifidobacterium spp., Lactobacillus spp., and gram-positive cocci were significantly (P < or = 0.05) higher in treatments PFW and PF compared with the control and AB treatments. Treatments PFW and PF had significantly higher specific activities of alpha-galactosidase and beta-galactosidase compared with the control and AB treatments. In conclusion, probiotic treatment PFW displayed a growth-promoting effect that was comparable to avilamycin treatment. In addition, treatments PFW and PF modulated the composition and, to an extent, the activities of the cecal microflora, resulting in a significant probiotic effect.