Effects of feed additives on ileal mucosa-associated microbiota composition of broiler chickens.

The effects of dietary supplementation with 2 recently developed feed additives on the composition of the mucosa-associated microbiota of the ileum were studied in growing broiler chickens. A total of 48 male 1-d-old broiler chickens of the Cobb 500 strain were distributed in 4 treatments with 2 replicates of 6 birds each. The 2 additives tested were a di-d-fructose dianhydride­enriched caramel (FC) and the garlic derivative propyl propane thiosulfonate (PTS-O). Dietary treatments were a control (commercial diet with no additive), INU (20 g inulin/kg diet), CAR (20 g FC/kg diet), and GAR (90 mgPTS-O/kg diet). As a result of this study, inulin supplementation resulted in lower (P < 0.05) and FC feeding resulted in higher (P < 0.05) Blautia coccoides/Eubacterium rectale log10 number of copies respect to controls. Higher (P < 0.05) bifidobacteria log10 number of copies with respect to the controls was determined in the ileal mucosa of birds fed the PTS-O­supplemented diet. Denaturing gradient gel electrophoresis and PCR analysis on Bifidobacterium spp. revealed the presence of Bifidobacterium longum, Bifidobacterium pseudolongum, and Bifidobacterium pseudocatenulatum in samples from chickens fed the control and the PTS-O­supplemented diet. Bifidobacterium longum was exclusively found in poultry fed the control diet, whereas B. pseudocatenulatum was found only in poultry fed the PTS-O­supplemented diet. This study showed that both PTS-O and FC were able to modulate the composition of the ileal mucosa-associated microbiota of growing broiler chickens. Finally, in addition to B. pseudolongum, the presence of B. longum and B. pseudocatenulatum, species not previously described in intestinal samples of broilers, was also demonstrated.

Correlations between changes in intestinal microbiota composition and performance parameters in broiler chickens.

Growing male Cobb broiler chickens were fed on diets supplemented with additives reported as able to influence intestinal microbiota composition. The diets used were a balanced commercial diet (no additive), inulin (20 g/kg), fructose caramel (FC, 20 g/kg) and the garlic derivative PTS-O (propyl propane thiosulfonate, 45 and 90 mg/kg diet). The composition of the intestinal microbiota was analysed by qPCR at different points of the intestinal tract, and a number of nutritional parameters were also determined. The relative amounts of bacteroides (bacteroides/total bacteria) in the ileal contents correlated (p < 0.05) positively with faecal NDF, ADF, hemicellulose and cellulose digestibility. The relative amounts of Escherichia-Shigella (Escherichia-Shigella/total bacteria) in the crop contents correlated (p = 0.05) negatively with weight gain of broilers. Faecal N digestibility correlated (p < 0.05) negatively with total bacteria in the ileal contents of chickens. The relative amounts of Escherichia-Shigella (Escherichia-Shigella/total bacteria) in the caecal contents correlated (p = 0.05) negatively with faecal fat digestibility of broilers. Total bacteria in ileal or caecal contents of growing chickens correlated (p < 0.05) negatively with ileal N digestibility. The results here reported suggest that positive or negative correlations can be found between performance parameters and changes in intestinal microbiota composition of growing broiler chickens.

Effects of inulin and di-D-fructose dianhydride-enriched caramels on intestinal microbiota composition and performance of broiler chickens.

In vitro and in vivo experiments were designed to evaluate the effectiveness of laboratory-made di-d-fructose dianhydride (DFA)-enriched caramels. The DFA-enriched caramels were obtained from d-fructose (FC), d-fructose and sucrose (FSC), or d-fructose and ß-cyclodextrin (FCDC). In the in vitro experiment, raftilose and all caramels increased (P<0.05) l-lactate concentration and decreased (P<0.05) pH. Total short-chain fatty acid concentration was higher (P<0.05) than controls in tubes containing raftilose, FSC, FCDC and commercial sucrose caramel (CSC). Raftilose, and all caramels tested except FSC and FC (1%), increased (P<0.01) lactobacilli log10 number of copies compared with the non-additive control. FSC, FCDC and CSC increased (P<0.01) the bifidobacteria number of copies as compared with controls. All additives, except FCDC, decreased (P<0.01) Clostridium coccoides/Eubacterium rectale log number of copies. Compared with controls, raftilose, FC and CSC led to lower (P<0.01) Escherichia-Shigella and enterobacteria. For the in vivo experiment, a total of 144 male 1-day-old broiler chickens of the Cobb strain were randomly assigned to one of the three dietary treatments for 21 days. Dietary treatments were control (commercial diet with no additive), inulin (20 g inulin/kg diet) and FC (20 g FC/kg diet). Final BW of birds fed FC diet was higher (P<0.01) than controls or inulin-fed birds, although feed: gain values were not different. Feed intake of chickens fed FC was higher (P<0.01) than that of inulin-fed birds but not statistically different from controls. Crop pH values were lower (P<0.01) in birds fed FC diet as compared with control diet, with inulin-fed chickens showing values not different from control- or FC-fed birds. Lower (P<0.05) lactobacilli number of copies was determined in the crop, ileum and caeca of birds fed the inulin diet compared with the control diet. Inulin supplementation also resulted in lower (P<0.05) C. coccoides/E. rectale, bacteroides and total bacteria in caecal contents. Addition of FC to broiler diets gave place to lower (P<0.05) enterobacteria and Escherichia-Shigella in crop and caecal contents compared with controls. The bacteroides number of copies increased (P<0.05) as compared with controls in the ileum, but decreased (P<0.05) in the caeca of chickens fed the FC diet. Energy, ADF, NDF and non-starch polysaccharides faecal digestibilities were greater (P<0.05) than controls in chickens fed diets containing inulin or FC. Fat digestibility was higher (P<0.05) in FC-fed birds compared with controls or inulin-fed chickens. In conclusion, DFA-enriched caramels tested here, particularly FC, may represent a type of new additives useful in poultry production.

Garlic derivative propyl propane thiosulfonate is effective against broiler enteropathogens in vivo.

Two experiments were carried out to study the effects of dietary supplementation with the garlic (Allium sativum)-derived product propyl propane thiosulfonate (PTS-O) on the intestinal log(10) number of copies of enteropathogens in broiler chickens, together with their intestinal morphology and growth performance. The additive had no significant effect on feed intake at any dose assayed. In experiment 1 (1 to 21 d of age), the BW of chickens fed on 45 mg of PTS-O/kg of diet was higher (P < 0.01) than that of controls. Birds fed on diets containing 45 and 90 mg of PTS-O/kg of diet had improved (P < 0.01) feed:gain ratios compared with the controls at 21 d of age. Ileal villus height, width and surface area, mucosal thickness, and muscular layer thickness were considerably greater (P < 0.01) than control values in chickens fed 90 mg of PTS-O/kg of diet. The Clostridium perfringens log(10) number of counts was not significantly affected at any dose assayed. The inclusion of PTS-O at both concentrations (45 and 90 mg/kg of diet) resulted in lower (P < 0.01) log(10) number of copies of ileal Salmonella spp. and crop enterobacteria and Escherichia coli. The inclusion of 90 mg of PTS-O/kg of diet also resulted in lower (P < 0.01) enterobacteria and E. coli log(10) numbers of copies in the ileal and cecal contents, respectively. The number of copies of Campylobacter jejuni was not significantly affected. In experiment 2 (15 to 28 d of age), lower (P < 0.01) log(10) number of copies of Salmonella spp. and C. jejuni were determined in the ileal contents of chickens fed on diets containing 135 mg of PTS-O/kg of diet. The addition of 90 mg of PTS-O/kg of diet lowered (P < 0.01) only the number of copies of ileal Salmonella spp. This investigation confirmed previous in vitro data and showed that PTS-O lowered the intestinal numbers of enteropathogens and improved the ileal histological structure and productive parameters of broilers.

Morphology and enzymatic activity of the small intestinal mucosa of Iberian pigs as compared with a lean pig strain.

Castrated male Iberian (n = 12) and Landrace × Large White (n = 12) pigs were used to study histological structure and enzymatic activity in the small intestine at 3 points of the productive cycle (BW = 15, 50, and 115 kg). Both strains were fed the same cereal-based diets (DE = 2,799 kcal·kg(-1), and CP = 15%) throughout the entire experimental period. Differences (P < 0.05) in histometrical variables (villus height, width and surface, crypt depth, villus height/crypt depth relationship, mucosal thickness, muscular layer thickness, and number of goblet cells) were found among samples of small intestinal sections (duodenum, jejunum, and ileum) at the 3 productive stages studied. Also, differences (P < 0.05) in histometrical variables of small intestinal samples were found between Iberian and lean pigs at all productive stages, although these differences tended to disappear with age. Differences (P < 0.05) in enzymatic activities (lactase, sucrase, maltase, isomaltase, aminopeptidase, and alkaline phosphatase) of small intestinal samples were found between the different intestinal sections at all productive stages studied. Although differences (P < 0.05) in enzymatic activities of small intestinal samples were found between Iberian and lean pigs at all productive stages, values tended to equalize with pig age. We concluded that differences previously found in dietary nutritional utilization between Iberian and lean strains are likely not due to differences in intestinal absorption or hydrolytic capacity.

A new experimental model of isolated myocardial ischemic injury: ECG findings of acute isolated right ventricular ischemic injury.

A new experimental porcine model for creating selective ischemia of a specific part of the myocardium while the rest of the myocardium remains free of ischemia has been used to study the electrocardiographic (ECG) changes deriving from selective ischemia of the right ventricular (RV) free wall. A patch was stitched to the ventricle to produce selective myocardial ischemic injury. In a preliminary study of nine pigs, selective ischemia of the left ventricular free wall in five and of the RV free wall in four animals was induced, and a postmortem dye injection was performed to evaluate blood flow in the area of ischemia. In an ECG study of 20 pigs, the baseline ECG was recorded with use of the standard leads I-III, aVR, aVL, and aVF, left precordial leads (V1-V6), and leads V4R, V3H, and V4H and 1 hour after inducing ischemia, the ECG study was repeated. Our experimental model produced ischemic injury in which the location and surface area were known antemortem. In the 20 pigs, ST-segment changes were recorded in leads V1-V3, V3H, and V4H. In only four pigs (20%) was ST-segment elevation recorded in lead V4R. The results show that the ECG signs of selective ischemia of the RV free wall may imitate the signs of anterior or anterolateral infarction of the left ventricle. In this study, elevation of the ST-segment in lead V4R was not pathognomonic of for RV ischemia. This model is a new tool for studying hemodynamic and ECG changes of selective univentricular or biventricular ischemic injury.