K. Fegeros

Effects of probiotic inclusion levels in broiler nutrition on growth performance, nutrient digestibility, plasma immunoglobulins, and cecal microflora composition

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.

Effect of dietary oregano oil supplementation on lamb meat characteristics.

The effect of dietary oregano essential oil supplementation on lamb meat characteristics was investigated. Eight male and eight female Chios lambs were divided into two equal groups. The first group was fed with the control diet consisting of concentrated feed and alfalfa hay, whereas the second group consumed the same diet, the only difference being that the concentrated feed was uniformly sprayed with oregano essential oil (1ml/kg). Duration of the experimental period was two months. No differences were observed after oregano essential oil supplementation in final body weight (kg), body weight gain (g) and carcass yield (%). Tenderness of longissimus thoracis muscle, expressed as sarcomere length and shear force value, was not influenced by the treatment, whereas pH and colour parameters (yellowness-redness) appeared to increase (P<0.05). Moreover, results showed that dietary incorporation of oregano essential oil exerted strong antioxidant effects retarding lipid oxidation (MDA formation) in meat during refrigerated and long-term frozen storage (P<0.001).

Use of Trichosporon mycotoxinivorans to suppress the effects of ochratoxicosis on the immune system of broiler chicks

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 μg/kg), OTA plus TRM at three inclusion rates (104 CFU/g of feed, 105 CFU/g, 106 CFU/g) and TRM alone at 105 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 μM): 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 30 d before the immunosuppressive effect became apparent. The dietary inclusion of TRM completely blocked the detrimental effects of OTA on several immune properties in broilers.

Effects of a multi-species probiotic on biomarkers of competitive exclusion efficacy in broilers challenged with Salmonella enteritidis

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 2⋅7 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.

Dietary probiotic inclusion level modulates intestinal mucin composition and mucosal morphology in broilers

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<0.001) with increasing probiotic feed inclusion level in 42-d-old broiler cecum, with treatment A not differing from treatment C (P≥0.05). Cecal villus height and crypt depth increased linearly (P=0.016 and P=0.003, respectively) with probiotic inclusion level, with treatment A having higher (P≤0.05) values only from treatment C. Mucus layer thickness increased linearly with probiotic inclusion level in duodenum at 14 d and 42 d (P=0.007 and P=0.030, respectively). Finally, mucus layer staining intensity was influenced (P<0.001) by villus fragment (i.e., tip, midsection, and base) but not from the treatment, age, and intestinal segment examined. As a conclusion, this study provides evidence that probiotic inclusion level affects intestinal mucin monosaccharide composition, mucus layer thickness, and intestinal morphology in broilers.

Influence of organic selenium supplementation on the accumulation of toxic and essential trace elements involved in the antioxidant system of chicken.

The aim of the study was to investigate the interactions between selenium (Se) and various trace elements, both toxic and essential, involved in the antioxidant system. A total of 128 day-old chicks (Gallus gallus, broilers) were used to investigate the effect of Se yeast supplementation on the accumulation of cadmium (Cd), copper (Cu) iron (Fe) and zinc (Zn). There were four replicates of four dietary treatments: T1 (basal diet with no added Se, analyzed to contain 0.21 mg kg−1), T2 (T1 with 0.15 mg kg−1 Se added), T3 (T1 with 0.3 mg kg−1 Se) and T4 (T1 with 3.0 mg kg−1 Se). At week 4 and 6, two chickens per replicate pen were sacrificed for whole blood, breast muscle and liver sampling. Samples were analyzed by ICP–MS. Supplementation with Se-yeast, not only increased Se concentration but also reduced Cd concentration in the tissues. Selenium was negatively correlated with Cd and positively correlated with Zn, Cu and Fe. Cadmium was negatively correlated with Zn and Cu. Zinc was positively correlated with Cu. Iron was negatively correlated with Cu and uncorrelated with Zn and Cd. The balance between Se, Cu, Fe and Zn is important for proper antioxidant defense since they are an integral part of various antioxidant enzymes.

The role of selenium in cadmium toxicity: interactions with essential and toxic elements

1. The study was part of a project designed to investigate if organic selenium (Se) can ameliorate the toxic effects of cadmium (Cd). The main objective of the present study was to investigate, in the chicken, the interactions between Se, Cd and the following elements: Sb, Ca, Cr, Co, Cu, Fe, Pb, Mg, Mn, Mo, Ni, V and Zn. 2. A total of 300 1-d-old chickens (broilers) were randomly distributed among 4 dietary treatments with 5 replicate pens per treatment. In T1, chickens were fed on a diet with 0·3 mg/kg added Se, without added Cd. In T2, chickens were fed on a diet with 0·3 mg/kg Se and 10 mg/kg Cd. In T3, chickens were fed on a diet with 0·3 mg/kg Se and 100 mg/kg of Cd added and in T4 treatment, chickens were fed on a diet with 3 mg/kg Se and 100 mg/kg Cd added. Se was added as Se-yeast. Cd was added as cadmium chloride (CdCl2). On d 28 and 42, two chickens per replicate pen were killed for collection of whole blood, liver, kidney and breast muscle samples. Samples were analysed by ICP-MS. The data were analysed using a multivariate linear model. 3. While low Cd concentrations in the diet led only to an increase of Cd concentration in the examined tissues, addition of high concentrations of Cd increased the concentration of Cd, Cu, Sb and V and decreased that of Se, Mn and Fe. Addition of high Se concentrations did not significantly reduce Cd concentration. 4. Prior to model application, correlations of 78 elements were noted, while after model application 39 correlations were noted. Most notably, Cd was correlated with Ca, Co, Cu and Mg, while Se was correlated with Mn. 5. The present study revealed several correlations between essential, probably essential and toxic elements illustrating the importance of the balance between pro-oxidants and antioxidants.

Supranutritional selenium level affects fatty acid composition and oxidative stability of chicken breast muscle tissue.

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 < 0.001, p = 0.023, p = 0.003 and p = 0.002, respectively) as the Se inclusion levels in the diets increased. At slaughter, a linear decrease in lipid oxidation (p = 0.019) was observed with Se addition, possibly attributed to the antioxidant properties of Se. Addition of supranutritional Se to chicken diets, at levels well below those causing toxicity, leads to production of Se-enriched meat, protection of health-promoting long-chain FA like C20:5n-3 and C22:6n-3 and protection of meat quality from oxidation at day 1 after slaughter.

Effects of Lactobacillus acidophilus on gut microflora metabolic biomarkers in fed and fasted rats

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 alpha-galactosidase, alpha-glucosidase and beta-glucosidase and higher activities of beta-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 beta-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.

Modulation of intestinal mucin composition and mucosal morphology by dietary phytogenic inclusion level in broilers

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.