The fecal biomarker ovotransferrin associates with broiler performance under field conditions.

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.

Guar gum as galactomannan source induces dysbiosis and reduces performance in broiler chickens and dietary ß-mannanase restores the gut homeostasis.

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. ß-Mannanases are enzymes that can hydrolyze ß-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 ß-mannanase supplementation, on the microbiota composition, in an attempt to describe the potential role of the intestinal microbiota in ß-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 + ß-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 ß-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 ß-mannanase supplementation restored the main microbial shifts induced by guar gum to levels of the control group. In addition, the ß-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. ß-Mannanase supplementation restored the gut microbiota composition and zootechnical performance to control levels.

Broiler performance correlates with gut morphology and intestinal inflammation under field conditions.

Maintaining optimal gut health is a key driver for a well-performing broiler flock. Histology of intestinal sections and quantification of villus structure can be used to evaluate gut health. While these measurements have been used in experimental models to evaluate gut health, less is known about the associations of these parameters with performance in commercial broiler farms. The objective of the present study was to evaluate possible associations of intestinal villus structure and the inflammatory condition of the gut with Ross 308 broiler performance in 50 commercial farms. On day 28 of the production round, 20 randomly selected broilers per farm were weighed, euthanized, and a duodenal section was collected to determine villus length, crypt depth and the CD3+ T-lymphocytes area percentage (CD3+ %). We found a relatively low coefficient of variance (CV) for the villus length (between farms; 9.67%, within farms; 15.97%), while the CD3+ (%) had a high CV (between farms; 29.78%, within farms; 25.55%). At flock level, the CD3+ (%) was significantly correlated with the villus length (r = -0.334), crypt depth (r = 0.523) and the villus-to-crypt ratio (r = -0.480). The crypt depth was significantly correlated with the European production index (EPI) (r = -0.450) and feed conversion ratio (FCR) (r = 0.389). At broiler level, a significant association was found between the individual body weight (day 28), CD3+ (%) and villus-to-crypt ratio. These data thus show that gut villus structure is significantly associated with bird performance under commercial conditions. RESEARCH HIGHLIGHTSGut histology parameters vary between and within farms.Broiler performance is associated with gut morphology.

Chestnut tannins in broiler diets: performance, nutrient digestibility, and meat quality.

In 2 broiler trials, the effects of chestnut tannins on performance and meat quality (trial 1), and digestion (trial 2) were evaluated. In both trials, Ross 308 broilers received one of 2 basal diets: one basal diet contained corn and soy as main feed ingredients, while the challenge basal diet contained wheat, palm oil, and rapeseed meal. The composition of the basal diets was chosen to assess the interaction between chestnut tannins and diet composition. To both basal diets, chestnut tannins were added at 3 doses: 0 mg/kg (T-), 500 mg/kg (T+), or 2,000 mg/kg (T++), resulting in a total of 6 treatments. In trial 1, both basal diets containing 2,000 mg/kg chestnut tannins lowered broiler performance in grower and finisher phases. A tannin dose of 500 mg/kg had no effect on performance in either basal diet. Corn-based diets resulted in lower meat pH compared to wheat diets. Further, addition of chestnut tannins resulted in increased meat pH, and caused proportionally a lower meat drip loss and shear force for both basal diets. During the digestibility study (trial 2), blood was also collected. None of the treatments affected digestibility or blood parameters (glucose, non-esterified fatty acids, and triacylglycerols). Malondialdehyde (MDA) was measured in plasma to assess antioxidative properties of chestnut tannins. In wheat diets, chestnut tannins significantly lowered plasma MDA demonstrating its antioxidative nature. Regarding gut health, crypt depth decreased proportionally with the dosage of chestnut tannins in both basal diets with significantly shallower crypts for the wheat diets compared to the corn diets. Relative intestinal growth was stimulated in the wheat diets proportionally to the tannin dose based on the larger relative gut length. In conclusion, chestnut tannins did not influence digestive metabolism, yet they lowered performance at higher doses regardless of feed ingredients used in the diet. Tannins positively affected meat quality and when added to wheat diets, intestinal growth was stimulated and the antioxidative status of the broilers improved.

Effect of vitamin E level and dietary zinc source on performance and intestinal health parameters in male broilers exposed to a temperature challenge in the finisher period.

The objective of this study was to evaluate the interaction of zinc source (ZnSO4 vs. zinc amino acid complex) and vitamin E level (50 IU vs. 100 IU) on performance and intestinal health of broilers exposed to a temperature challenge in the finisher period. A total of 1224 day old male Ross 308 broilers were randomly distributed among 4 dietary treatments (9 replicates per treatment). Dietary treatments were organized in a 2 × 2 factorial arrangement: two sources of zinc, 60 mg/kg of Zn as ZnSO4 .7H2 O or 60 mg/kg of Zn as zinc amino acid complexes (ZnAA) combined with two levels of vitamin E (50 or 100 IU/kg). Zinc and vitamin E were added to a wheat/rye-based diet that was designed to create a mild nutritional challenge. From day 28 until day 36 (finisher period), all birds were subjected to chronic cyclic high temperatures (32°C ± 2°C and RH 55-65% for 6 h daily). The combination of ZnAA and 50 IU/kg of vitamin E improved weight gain in the starter (day 0-10), finisher (day 28-36) and overall period (day 0-36) and feed conversion ratio in the starter (day 0-10) and finisher phase (day 28-36). Providing Zn as ZnAA significantly improved villus length and villus/crypt ratio in the starter, grower and finisher period and decreased infiltration of T-lymphocytes and ovotransferrin leakage in the finisher period. In conclusion, providing broilers with a diet supplemented with ZnAA and a vitamin E level of 50 IU/kg, resulted in better growth performance as compared to all other dietary treatments. Interestingly, under the conditions of this study, positive effects of ZnAA on performance did not occur when vitamin E was supplemented at 100 IU/kg in feed. Moreover, providing zinc as zinc amino acid complex improved intestinal health.

Calcination Improves the In Vivo Efficacy of a Montmorillonite Clay to Bind Aflatoxin G1 in Broiler Chickens: A Toxicokinetic Approach.

The goal of this study was to investigate the toxicokinetic characteristics of aflatoxin G1 (AFG1) in broiler chickens and the effect of calcination of a Tunisian montmorillonite clay on the in vivo absorption of AFG1. In this study, broiler chickens were randomly distributed into four groups of 10 animals. Group 1 was administered AFG1 (2 mg/kg body weight (BW)) by single intravenous injection (IV), group 2 received an intra-crop bolus (PO) of AFG1 without any clay, group 3 was dosed AFG1 PO together with an oral bolus of purified clay (CP), and group 4 received AFG1 PO with an oral bolus of calcined clay. A significant difference in the area under the curve (AUC0-t) was observed for group 4 (6.78 ± 4.24 h*ng/mL) in comparison with group 2 (12.83 ± 4.19 h*ng/mL). A significant reduction of the oral bioavailability of AFG1 was observed for group 4 (7.61 ± 4.76%) compared with group 2 (14.40 ± 4.70%), while no significant effect was observed of CP. In this experiment, no phase I nor phase II metabolites of AFG1 were observed. These findings confirm that calcination of the purified montmorillonite clay enhances the adsorption of AFG1 in the gastrointestinal tract after oral administration, thereby reducing its bioavailability, thus reducing its toxic effects.

Dietary zinc source impacts intestinal morphology and oxidative stress in young broilers.

Zinc is an essential nutritional trace element for all forms of life as it plays an important role in numerous biological processes. In poultry, zinc is provided by in-feed supplementation, mainly as zinc oxide or zinc sulfate. Alternatively zinc can be supplemented as organic sources, which are characterized by using an organic ligand that may be an amino acid, peptide, or protein to bind zinc and have a higher bioavailability than inorganic zinc sources. There are limited number of studies directly comparing the effects of inorganic vs. organic zinc sources on performance and intestinal health in broilers. Therefore, a digestibility and a performance study were conducted to evaluate and compare the effect of an amino acid-complexed zinc source vs. an inorganic zinc source on intestinal health. The experiment consisted of 2 treatments: either a zinc amino acid complex or zinc sulfate was added to a wheat-rye based diet at 60 ppm Zn, with 10 replicates (34 broilers per pen) per treatment. Effects on performance, intestinal morphology, microbiota composition, and oxidative stress were measured. Supplementing zinc amino acid complexes improved the zinc digestibility coefficient as compared to supplementation with zinc sulfate. Broilers supplemented with zinc amino acid complexes had a significantly lower feed conversion ratio in the starter phase compared to birds supplemented with zinc sulfate. A significantly higher villus length was observed in broilers supplemented with zinc amino acid complexes at days 10 and 28. Supplementation with zinc amino acid complexes resulted in a decreased abundance of several genera belonging to the phylum of Proteobacteria. Plasma malondialdehyde levels and glutathione peroxidase activity showed an improved oxidative status in broilers supplemented with zinc amino acid complexes. In conclusion, zinc supplied in feed as amino acid complex is more readily absorbed, potentially conferring a protective effect on villus epithelial cells in the starter phase.

Effect of in feed administration of different butyrate formulations on Salmonella Enteritidis colonization and cecal microbiota in broilers.

Butyrate has been used extensively as a feed additive to improve gut health and to decrease Salmonella colonization in poultry. Salmonella mainly colonizes the ceca so butyrate concentrations should be increased in this gut segment. Discrepancies on the effects of butyrate on Salmonella colonization, described in the scientific literature, could thus be due to butyrate release location effects. In this study, newly developed butyrate formulations were evaluated for their effect on cecal butyrate concentrations and on colonization by Salmonella Enteritidis. In a first trial, broilers were randomly allocated to 7 dietary treatment groups with formulations based on different approaches to modify the butyrate release profile: release from wax matrices based on diffusion/erosion; micropellets supposedly release butyrate around pH 7 in the colon; tributyrin is based on the hydrolysis of esters in the small intestine. Fat-protected butyrate was included as a reference, because of its known effect on reduction of Salmonella colonization. Four days after infection, the number of cfu Salmonella per g cecal content and spleen were determined. Butyrate formulations in a wax matrix significantly reduced the Salmonella colonization in cecal content. In a second trial, wax and fat-protected butyrate treatments were replicated and results from the first trial were confirmed. Compared to the control group a higher proportion of butyrate concentration was observed in ceca for those groups with reduced Salmonella colonization. This was associated with a beneficial shift in the cecal microbiota. In conclusion, formulations that increase cecal butyrate concentrations are superior in protecting against Salmonella Enteritidis colonization.

Amorphous cellulose feed supplement alters the broiler caecal microbiome.

The grains that form the basis of most commercial chicken diets are rich in cellulose, an unbranched ß-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.

In-feed resin acids reduce matrix metalloproteinase activity in the ileal mucosa of healthy broilers without inducing major effects on the gut microbiota.

The chicken gut is constantly exposed to harmful molecules and microorganisms which endanger the integrity of the intestinal wall. Strengthening intestinal mucosal integrity is a key target for feed additives that aim to promote intestinal health in broilers. Recently, dietary inclusion of resin-based products has been shown to increase broiler performance. However, the mode of action is still largely unexplored. Coniferous resin acids are known for their anti-microbial, anti-inflammatory and wound-healing properties, all properties that might support broiler intestinal health. In the current study, the effect of pure resin acids on broiler intestinal health was explored. Ross 308 broilers were fed a diet supplemented with coniferous resin acids for 22 days, after which the effect on both the intestinal microbiota as well as on the intestinal tissue morphology and activity of host collagenases was assessed. Dietary inclusion of resin acids did not alter the morphology of the healthy intestine and only minor effects on the intestinal microbiota were observed. However, resin acids-supplementation reduced both duodenal inflammatory T cell infiltration and small intestinal matrix metalloproteinase (MMP) activity towards collagen type I and type IV. Reduced breakdown of collagen type I and IV might indicate a protective effect of resin acids on intestinal barrier integrity by preservation of the basal membrane and the extracellular matrix. Further studies are needed to explore the protective effects of resin acids on broiler intestinal health under sub-optimal conditions and to elaborate our knowledge on the mechanisms behind the observed effects.

In-feed bambermycin medication induces anti-inflammatory effects and prevents parietal cell loss without influencing Helicobacter suis colonization in the stomach of mice.

The minimum inhibitory concentration of bambermycin on three porcine Helicobacter suis strains was shown to be 8 µg/mL. The effect of in-feed medication with this antibiotic on the course of a gastric infection with one of these strains, the host response and the gastric microbiota was determined in mice, as all of these parameters may be involved in gastric pathology. In H. suis infected mice which were not treated with bambermycin, an increased number of infiltrating B-cells, T-cells and macrophages in combination with a Th2 response was demonstrated, as well as a decreased parietal cell mass. Compared to this non-treated, infected group, in H. suis infected mice medicated with bambermycin, gastric H. suis colonization was not altered, but a decreased number of infiltrating T-cells, B-cells and macrophages as well as downregulated expressions of IL-1ß, IL-8M, IL-10 and IFN-γ were demonstrated and the parietal cell mass was not affected. In bambermycin treated mice that were not infected with H. suis, the number of infiltrating T-cells and expression of IL-1ß were lower than in non-infected mice that did not receive bambermycin. Gastric microbiota analysis indicated that the relative abundance of bacteria that might exert unfavorable effects on the host was decreased during bambermycin supplementation. In conclusion, bambermycin did not affect H. suis colonization, but decreased gastric inflammation and inhibited the effects of a H. suis infection on parietal cell loss. Not only direct interaction of H. suis with parietal cells, but also inflammation may play a role in death of these gastric acid producing cells.

Valeric acid glyceride esters in feed promote broiler performance and reduce the incidence of necrotic enteritis.

Valeric acid is a C5 fatty acid, naturally produced in low concentrations by specific members of the microbiota of the lower intestinal tract. Effects of valeric acid on intestinal health have been poorly investigated. Valeric acid derivatives can be produced as glyceride esters and added to broiler feed. In the current study, experiments were carried out to evaluate the effect of valeric acid glycerides (GVA) on growth performance, on the morphology of the small intestinal mucosa and on protection against necrotic enteritis. In a first feeding trial, Ross-308 chicks were randomly divided into 2 dietary treatment groups and fed either a non-supplemented diet or a diet supplemented with GVA (1.5 g/kg). In the GVA supplemented group, the feed conversion ratio was significantly decreased during the entire trial period (D1-37). In a second trial, gut wall morphology was evaluated. In broilers fed a GVA-containing diet at 5 g/kg, the villus height/crypt depth ratio in the jejunum was significantly increased (P ≤ 0.05), and the crypt depth was significantly decreased at 28 d. In a third trial, immunohistochemistry showed that the density of glucagon-like peptide-2 immunoreactive cells in jejunal and ileal villi from broilers supplemented with GVA (5 g/kg) was significantly increased (P ≤ 0.05) on d 10. In a necrotic enteritis challenge model, a significant reduction of the number of birds with necrotic lesions was found at d 21, using in-feed supplementation of low and high regimen of GVA. These data show that GVA supplementation to broiler feed can decrease the feed conversion, positively affect the morphology of the small intestinal mucosa, increase the density of glucagon-like peptide-2 producing enteroendocrine cells, and reduce the incidence of necrotic enteritis, making GVA a valuable candidate feed additive for broilers.

Oral glutathione supplementation drastically reduces Helicobacter-induced gastric pathologies.

Helicobacter (H.) suis causes gastric pathologies in both pigs and humans. Very little is known on the metabolism of this bacterium and its impact on the host. In this study, we have revealed the importance of the glutamate-generating metabolism, as shown by a complete depletion of glutamine (Gln) in the medium during H. suis culture. Besides Gln, H. suis can also convert glutathione (GSH) to glutamate, and both reactions are catalyzed by the H. suis γ-glutamyltranspeptidase (GGT). Both for H. pylori and H. suis, it has been hypothesized that the degradation of Gln and GSH may lead to a deficiency for the host, possibly initiating or promoting several pathologies. Therefore the in vivo effect of oral supplementation with Gln and GSH was assessed. Oral supplementation with Gln was shown to temper H. suis induced gastritis and epithelial (hyper)proliferation in Mongolian gerbils. Astonishingly, supplementation of the feed with GSH, another GGT substrate, resulted in inflammation and epithelial proliferation levels returning to baseline levels of uninfected controls. This indicates that Gln and GSH supplementation may help reducing tissue damage caused by Helicobacter infection in both humans and pigs, highlighting their potential as a supportive therapy during and after Helicobacter eradication therapy.

Effects of Helicobacter suis γ-glutamyl transpeptidase on lymphocytes: modulation by glutamine and glutathione supplementation and outer membrane vesicles as a putative delivery route of the enzyme.

Helicobacter (H.) suis colonizes the stomach of the majority of pigs as well as a minority of humans worldwide. Infection causes chronic inflammation in the stomach of the host, however without an effective clearance of the bacteria. Currently, no information is available about possible mechanisms H. suis utilizes to interfere with the host immune response. This study describes the effect on various lymphocytes of the γ-glutamyl transpeptidase (GGT) from H. suis. Compared to whole cell lysate from wild-type H. suis, lysate from a H. suis ggt mutant strain showed a decrease of the capacity to inhibit Jurkat T cell proliferation. Incubation of Jurkat T cells with recombinantly expressed H. suis GGT resulted in an impaired proliferation, and cell death was shown to be involved. A similar but more pronounced inhibitory effect was also seen on primary murine CD4(+) T cells, CD8(+) T cells, and CD19(+) B cells. Supplementation with known GGT substrates was able to modulate the observed effects. Glutamine restored normal proliferation of the cells, whereas supplementation with reduced glutathione strengthened the H. suis GGT-mediated inhibition of proliferation. H. suis GGT treatment abolished secretion of IL-4 and IL-17 by CD4(+) T cells, without affecting secretion of IFN-γ. Finally, H. suis outer membrane vesicles (OMV) were identified as a possible delivery route of H. suis GGT to lymphocytes residing in the deeper mucosal layers. Thus far, this study is the first to report that the effects on lymphocytes of this enzyme, not only important for H. suis metabolism but also for that of other Helicobacter species, depend on the degradation of two specific substrates: glutamine and reduced glutatione. This will provide new insights into the pathogenic mechanisms of H. suis infection in particular and infection with gastric helicobacters in general.

Antibacterial therapeutics for the treatment of chytrid infection in amphibians: Columbus's egg?

BACKGROUND: The establishment of safe and effective protocols to treat chytridiomycosis in amphibians is urgently required. In this study, the usefulness of antibacterial agents to clear chytridiomycosis from infected amphibians was evaluated. RESULTS: Florfenicol, sulfamethoxazole, sulfadiazine and the combination of trimethoprim and sulfonamides were active in vitro against cultures of five Batrachochytrium dendrobatidis strains containing sporangia and zoospores, with minimum inhibitory concentrations (MIC) of 0.5-1.0 µg/ml for florfenicol and 8.0 µg/ml for the sulfonamides. Trimethoprim was not capable of inhibiting growth but, combined with sulfonamides, reduced the time to visible growth inhibition by the sulfonamides. Growth inhibition of B. dendrobatidis was not observed after exposure to clindamycin, doxycycline, enrofloxacin, paromomycin, polymyxin E and tylosin. Cultures of sporangia and zoospores of B. dendrobatidis strains JEL423 and IA042 were killed completely after 14 days of exposure to 100 µg/ml florfenicol or 16 µg/ml trimethoprim combined with 80 µg/ml sulfadiazine. These concentrations were, however, not capable of efficiently killing zoospores within 4 days after exposure as assessed using flow cytometry. Florfenicol concentrations remained stable in a bathing solution during a ten day period. Exposure of Discoglossus scovazzi tadpoles for ten days to 100 µg/ml but not to 10 µg florfenicol /ml water resulted in toxicity. In an in vivo trial, post metamorphic Alytes muletensis, experimentally inoculated with B. dendrobatidis, were treated topically with a solution containing 10 µg/ml of florfenicol during 14 days. Although a significant reduction of the B. dendrobatidis load was obtained, none of the treated animals cleared the infection. CONCLUSIONS: We thus conclude that, despite marked anti B. dendrobatidis activity in vitro, the florfenicol treatment used is not capable of eliminating B. dendrobatidis infections from amphibians.

An update on alternatives to antimicrobial growth promoters for broilers.

Livestock performance and feed efficiency are closely interrelated with the qualitative and quantitative microbial load of the animal gut, the morphological structure of the intestinal wall and the activity of the immune system. Antimicrobial growth promoters have made a tremendous contribution to profitability in intensive husbandry, but as a consequence of the increasing concern about the potential for antibiotic resistant strains of bacteria, the European Commission decided to ban all commonly used feed antibiotics. There are a number of non-therapeutic alternatives, including enzymes, (in)organic acids, probiotics, prebiotics, etheric oils and immunostimulants. Their efficacy and mode of action are briefly described in this review.

The cereal type in feed influences gut wall morphology and intestinal immune cell infiltration in broiler chickens.

In broiler chickens, a diet where the major cereal types are wheat, rye and/or barley has a lower digestibility compared with a diet in which maize is the major cereal type. In the present study, the effects of two different dietary cereal types, maize v. wheat/rye, on host factors (inflammation and gut integrity) and gut microbiota composition were studied. In addition, the effects of low-dose Zn-bacitracin supplementation were examined. Broilers given a wheat/rye-based diet showed more villus fusion, a thinner tunica muscularis, more T-lymphocyte infiltration, higher amount of immune cell aggregates in the mucosa, more and larger goblet cells and more apoptosis of epithelial cells in the mucosa than those given a maize-based diet. Adding Zn-bacitracin generally reversed these alterations. The microbiota composition was analysed by the use of terminal-restriction fragment length polymorphism, showing changes in the microbiota composition depending on the cereal type used in the diets. The effect of the change of cereal type on the gut microbiota composition was larger than that of Zn-bacitracin supplementation. In conclusion, a wheat/rye-based diet evoked mucosal damage, an alteration in the composition of the microbiota and an inflammatory bowel type of condition.

Short-chain fatty acids and L-lactate as feed additives to control Campylobacter jejuni infections in broilers.

The usefulness of butyrate, acetate, propionate and l-lactate for the control of Campylobacter jejuni infections in broilers was assessed. For this purpose, the effect of these acids on the growth of C. jejuni in broth and intestinal mucous was determined, as well as their influence on the invasiveness of C. jejuni in intestinal epithelial cells. From these in vitro obtained results, one acid was retained for use as a feed additive in an in vivo trial. Butyrate was the most successful of the short-chain fatty acids, with 12.5 mM being bactericidal for C. jejuni at pH 6.0. Propionate and acetate had a bacteriostatic effect at 50 mM. None of the short-chain fatty acids had a bactericidal effect at pH 7.5 at a maximum concentration of 50 mM. Mucous increased the minimum bactericidal concentration of butyrate, but not the bacteriostatic concentrations of propionate or acetate. When C. jejuni was incubated in growth subinhibitory concentrations of butyrate, acetate or propionate or 25 mM L-lactate, no alteration in the invasive capabilities of C. jejuni in Caco-2 cells was observed. The addition of butyrate-coated micro-beads to the feed was unsuccessful to reduce C. jejuni caecal colonization in a seeder model using 2-week-old broilers. In conclusion, despite the marked bactericidal effect of butyrate towards C. jejuni in vitro, butyrate-coated micro-beads do not protect broilers from caecal colonization with C. jejuni in the applied test conditions. This might be partially ascribed to the protective effect of mucous and the rapid absorption of butyrate by the enterocytes.

Can autologous thrombin with a rest fraction of ethanol be used safely for activation of concentrated autologous platelets applied on nerves?

The use of concentrated platelet-rich plasma (cPRP) as a source of growth factors is reported to be beneficial for an enhanced osteogenesis in spine surgery. Today both bovine and autologous thrombin is used for activating the platelets and thus releasing the growth factors. In order to prevent transmission of organisms and the development of antibodies, autologous thrombin seems to be the logical choice. However, the preparation of autologous thrombin is cumbersome and consumes a part of the cPRP. In order to overcome this problem, a commercial autologous thrombin kit (activAT, Dideco, a Sorin Group company, Italy) has been developed which produces autologous thrombin out of platelet-poor plasma. A possible disadvantage of this kit could be the rest fraction of 1.18% of ethanol in the platelet gel. In a pig model, the influence of different ethanol concentrations on the ischiadic nerve was studied. The study consisted out of four groups; a control group (n=6), a group with platelet gel 0% ethanol (n=6), a group with platelet gel 1.18% ethanol (n=6) and a group with platelet gel 3.8% ethanol (n=7). In all the groups, the ischiadic nerve was dissected and the myelin sheet opened after which the wound was closed (control group) or one of the three therapies was applied. After 12 h, the animal was sacrificed and the ischiadic nerve was submitted for histological examination. Myelin sheaths appeared normal in all cases. No axonal swelling was observed. No statistically significant differences were observed for neutrophilic and eosinophilic infiltration nor for collagen necrosis between groups. Platelet gel prepared by the use of a commercial autologous thrombin kit and containing 1.18% of ethanol can be safely used on nerves.