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Broilers often suffer from subclinical intestinal health problems of ill-defined etiology, which have a negative impact on performance. Macroscopic and microscopic evaluations can be used to monitor intestinal health, but because these are subjective and time-consuming, respectively, objective and easy-to-measure biomarkers are urgently needed. Fecal biomarkers can potentially be used as noninvasive, objective measures to evaluate gut health in broilers. The aim of the current study was to evaluate ovotransferrin (OVT) as a biomarker in fecal/colonic samples derived from broilers from 27 industrial farms by investigating associations between OVT, broiler performance and gut histology parameters. Eight chickens per farm were randomly selected, weighed and euthanized on d 28 of the production round. A duodenal section was collected to measure the intestinal villus structure (villus length, crypt depth) and the inflammatory status of the gut (CD3 + T-lymphocytes area percentage). The coefficient of variation for the OVT (between farms; 83.45%, within farms; 95.13%) was high compared to the villus length (between farms; 10.91%, within farms; 15.48%), crypt depth (between farms; 15.91%, within farms; 14.10%), villus-to-crypt ratio (between farms; 22.08%, within farms; 20.53%), and CD3 + (between farms; 36.38%, within farms; 26.13%). At farm level, colonic OVT was significantly associated with the average slaughter weight (P = 0.005), daily weight gain (P = 0.007) and the European production index (EPI) (P = 0.009). At broiler level, significant associations were found between colonic OVT and the villus length (P = 0.044) and between the colonic OVT and villus-to-crypt ratio (P = 0.050). These results thus show that quantifying OVT in colon can have merit for evaluation of intestinal health in broilers under field conditions., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Galactomannans are abundant nonstarch polysaccharides in broiler feed ingredients. In broilers, diets with high levels of galactomannans have been associated with innate immune response stimulation, poor zootechnical performance, nutrient and lipid absorption, and excessive digesta viscosity. However, data about its effects on the gut microbiome are scarce. β-Mannanases are enzymes that can hydrolyze β-mannans, resulting in better nutrient utilization. In the current study, we have evaluated the effect of guar gum, a source of galactomannans, supplemented to broiler diets, either with or without β-mannanase supplementation, on the microbiota composition, in an attempt to describe the potential role of the intestinal microbiota in β-mannanase-induced gut health and performance improvements. One-day-old broiler chickens (n = 756) were randomly divided into 3 treatments: control diet, guar gum-supplemented diet (1.7%), or guar gum-supplemented diet + β-mannanase (Hemicell 330 g/ton). The zootechnical performance, gut morphometry, ileal and cecal microbiome, and short-chain fatty acid concentrations were evaluated at different time points. The guar gum supplementation decreased the zootechnical performance, and the β-mannanase supplementation restored performance to control levels. The mannan-rich diet-induced dysbiosis, with marked effects on the cecal microbiota composition. The guar gum-supplemented diet increased the cecal abundance of the genera Lactobacillus, Roseburia, Clostridium sensu stricto 1, and Escherichia-Shigella, and decreased Intestinimonas, Alistipes, Butyricicoccus, and Faecalibacterium. In general, dietary β-mannanase supplementation restored the main microbial shifts induced by guar gum to levels of the control group. In addition, the β-mannanase supplementation reduced cecal isobutyric, isovaleric, valeric acid, and branched-chain fatty acid concentrations as compared to the guar gum-supplemented diet group, suggesting improved protein digestion and reduced cecal protein fermentation. In conclusion, a galactomannan-rich diet impairs zootechnical performance in broilers and results in a diet-induced dysbiosis. β-Mannanase supplementation restored the gut microbiota composition and zootechnical performance to control levels., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Intestinal integrity losses have been identified as a main driver for poor performance in broilers. The oral administration of markers such as iohexol is a major asset for measuring intestinal permeability (IP) alterations. The aim of the current study was to evaluate oral iohexol administration and serum levels as a quantitative measure for IP in Ross 308 broilers and to identify possible associations with histologic measurements. A total of 40, day-old broiler chickens were randomly divided into 4 groups of 10 broilers and a coccidiosis model was used to induce IP. Three challenge groups received a mixture of different field strains and concentrations of Eimeria acervulina and Eimeria maxima at d 16, and 1 group operated as an uninfected control group. On d 20, 5 birds per group were orally administered the permeability marker iohexol at a dose of 64.7 mg/kg body weight and blood was taken 60 min after the oral gavage. On d 21 these 5 birds per group were euthanized. On d 21, 5 other birds per group were given iohexol where after blood was taken. These birds were euthanized on d 22. During necropsy, birds were scored for coccidiosis lesions and a duodenal segment was taken for histology. The Eimeria challenge had a significant impact on the villus length, crypt depth, villus-to-crypt ratio and CD3 + T-lymphocytes area percentage. Challenged birds had a significant higher concentration of serum iohexol on both sampling days, as compared to the uninfected controls. A significant correlation could be found between the serum iohexol concentration and the histologic parameters (villus length, crypt depth and villus-to-crypt ratio) on the first sampling day. This suggests that iohexol may be used as a gut permeability marker in broilers under Eimeria challenge., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The grains that form the basis of most commercial chicken diets are rich in cellulose, an unbranched β-1,4-linked D-glucopyranose polymer, used as a structural molecule in plants. Although it is a predominant polysaccharide in cereal hulls, it is considered an inert non-fermentable fiber. The aim of the current study was to analyze the effect of in-feed supplementation of cellulose on the gut microbiota composition of broilers. Administration of cellulose to chickens, on top of a wheat-based diet, changed the caecal microbiota composition, as determined using pyrosequencing of the 16S rRNA gene. At day 26, a significantly (P < 0.01) higher relative abundance of the Alistipes genus was observed in the caeca of broilers fed the cellulose-supplemented diet, compared to animals fed the control diet. An in vitro batch fermentation assay showed a significant (P < 0.01) growth stimulation of Alistipes finegoldii in the presence of cellulose. In conclusion, in-feed supplementation of cellulose alters the microbiota composition at the level of the phylum Bacteroidetes, specifically the Alistipes genus. This suggests that cellulose is not essentially inert but can alter the gut micro-environment., (© 2019 Poultry Science Association Inc.)

Salmonella Enteritidis has developed the potential to contaminate eggs by surviving in the antimicrobial environment of the hen's egg white. This has led to a worldwide pandemic of foodborne salmonellosis infections in humans due to the consumption of contaminated eggs and egg-derived products. The molecular mechanisms of Salmonella Enteritidis egg white survival are not fully clear. Using in vivo expression technology and promoter-reporter fusions we showed that the promoter of the tolC gene, encoding the TolC outer membrane channel that is used by multidrug efflux pumps to export harmful molecules and to secrete bacterial products, is activated by egg white at the chicken body temperature. Using a Salmonella Enteritidis tolC deletion mutant we showed that TolC has an important role in egg white survival. Chromatographic separation techniques and subsequent testing of antimicrobial activities of separated egg white fractions led to the identification of ovotransferrin as the egg white antimicrobial factor which is capable of inhibiting growth of a tolC deletion strain but not the wild type strain. We provide evidence that TolC protects Salmonella Enteritidis against ovotransferrin-mediated growth inhibition in egg white., (© 2019 Poultry Science Association Inc.)

Carbohydrate-degrading multi-enzyme preparations (MEP) are used to improve broiler performances. Their mode of action is complex and not fully understood. In this study, we compared the effect of water-soluble fractions isolated at the pilot scale from wheat grain incubated with (WE) and without (WC) MEP. The fractions were incorporated in a wheat-based diet (0.1% w/w) to feed Ross PM3 broilers and compared with a non-supplemented control group (NC). The body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) until d 14 were determined. At d 14, ileal and cecal contents and tissue samples were collected from euthanized animals. The intestinal contents were used to measure the short-chain fatty acids (SCFA) concentration using gas chromatography and to determine the abundance and composition of microbiota using 16S sequencing. Villi length of ileal samples was measured, while L-cell and T-cell densities were determined using immuno-histochemistry. The MEP treatment increased the amount of water-soluble arabinoxylans (AX) and reduced their molecular weight while retaining their polymer behavior. The WE fraction significantly (P < 0.05) increased FI by 13.8% and BWG by 14.7% during the first wk post hatch when compared to NC. No significant effect on FCR was recorded during the trial. The WE increased the abundance of Enterococcus durans and Candidatus arthromitus in the ileum and of bacteria within the Lachnospiraceae and Ruminococcaceae families, containing abundant butyrate-producing bacteria, in the ceca. It also increased the concentration of SCFA in the ceca, decreased the T-lymphocyte infiltration in the intestinal mucosa, and increased the glucagon-like-peptide-2 (GLP-2)-producing L-cell density in the ileal epithelium compared with WC and NC. No significant effects were observed on villi length. These results showed that AX present in the WE fraction altered the microbiota composition towards butyrate producers in the ceca. Butyrate may be responsible for the reduction of inflammation, as suggested by the decrease in T-lymphocyte infiltration, which may explain the higher feed intake leading to improved animal growth., (© 2017 Poultry Science Association Inc.)

The non-starch polysaccharides (NSPs) in cell walls can act as a barrier for digestion of intracellular nutrients. This effect is called "cage effect." Part of the success of fibrolytic enzymes in broiler feed is assumed to be attributed to cage effect reduction. Further, changes in viscosity and potential prebiotic action should also be considered. The aim of this study was to gain insight into the relative importance of the cage effect in xylanase efficacy in broilers. Using a 2×2 factorial design, 24 pens with 30 Ross 308 male chicks were fed corn-soy based diets consisting of normal and freeze-thawed (5 d at -18°C) corn, both with and without xylanase. The freeze-thaw method was used to eliminate the cage effect, whereas a corn-based diet was used to exclude viscosity effects. Body weights (BW), feed intake (FI), and feed conversion ratio (FCR) were determined at d 13, 26, and 39. A balance study was executed at the end of the growing phase. These birds were euthanized at d 34 (non-fasted) to determine the viscosity of digesta, blood metabolites, intestinal morphology, and microbiota composition. During the finisher period, there was a significant interaction between enzyme supplementation and freeze-thawing for FCR, in which FCR was improved by freeze-thawed corn and tended to be improved by normal corn+enzyme compared with the control group. The improvement in performance (finisher period) of freeze-thawed corn and xylanase coincided with increased gut absorption of glucose (based on postprandial plasma concentrations) and increased number of Clostridiumcluster IV in the caecum, and agreed with the higher gut villus height. In addition, xylanase inclusion significantly increased the postprandial plasma glycine and triglycerides concentration, and led to elevated bacterial gene copies of butyryl CoA:acetate CoA-transferase, suggesting a prebiotic effect of xylanase addition through more than just the cage effect reduction. The applied model managed to rule out viscosity by using corn, and it was possible to isolate the cage effect by freeze-thawing the dietary corn., (© 2016 Poultry Science Association Inc.)

Dysbiosis or dysbacteriosis is defined as a shift in the intestinal microbiota composition resulting in an imbalance between beneficial and harmful bacteria. Since the ban on the use of growth-promoting antibiotics in animal feed in the EU, dysbiosis has emerged as a major problem in intensive animal production. Prebiotics and probiotics are currently under investigation as possible alternatives to growth-promoting antibiotics, as their mode of action is thought to be based largely on a modulation of the composition and function of the intestinal microbiota. In this review, we analyse the currently available data from both animal and human nutrition that document the potential and limitations of prebiotics and probiotics for the control of dysbiosis. An impressive number of empirical feeding trials have been carried out in healthy animals, yielding sometimes contradictory results. More in-depth studies have revealed the complexity of the interactions taking place in the lower intestinal tract, thus illustrating that pre- and probiotics cannot be a simple replacement for growth-promoting antibiotics. Although there are indications that the strategic use of pre- and probiotics can provide major benefits, there is still a lack of basic knowledge on the delicate interactions between the microbiota, the host and the feed components, which hampers the widespread use of these valuable feed additives.

The objective of this in vivo experiment was to evaluate the influence of arabinoxylooligosaccharides (AXOS) on shedding and colonization of Salmonella Enteritidis in broilers. Arabinoxylooligosaccharides, which are oligosaccharides derived from arabinoxylans by partial hydrolysis, have a beneficial effect on feed conversion ratios when added to broiler diets. Additionally, AXOS have been shown to promote the growth of bifidobacteria in the cecocolonic compartment of the gastrointestinal tract. To investigate the impact of AXOS on colonization of broilers with Salmonella, 224 one-day-old chicks were divided into 4 groups and given either unsupplemented feed or feed supplemented with 0.2% AXOS-3-0.25, 0.2% AXOS-9-0.25, or 0.4% AXOS-9-0.25 throughout the experiment. The AXOS-3-0.25 and AXOS-9-0.25 both have an ara-binose-to-xylose ratio of 0.25 and have an average degree of polymerization of 3 and 9, respectively. At 14 d posthatch, each animal was orally inoculated with 2.5 x 10(9) cfu of Salmonella Enteritidis. Cloacal swabs, taken at regular times, showed a significant reduction of Salmonella presence in the group given 0.4% AXOS-9-0.25 compared with the control group. This reduction was observed in the 1- to 11-d postinfection period. Colonization of the ceca as well as the translocation of Salmonella to the spleen was significantly reduced at 3 and 7 d postinfection in the 0.4% AXOS-9-0.25 group. A similar, although more moderate, decrease in colonization was observed in the group given 0.2% AXOS-9-0.25. It was concluded that dietary addition of AXOS provides dose-dependent protection against oral infections with Salmonella Enteritidis in poultry.