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Muramidases constitute a superfamily of enzymes that hydrolyse peptidoglycan (PGN) from bacterial cell walls. Recently, a fungal muramidase derived from Acremonium alcalophilum has been shown to increase broiler performance when added as a feed additive. However, the underlying mechanisms of action are not yet identified. Here, we investigated the hypothesis that this muramidase can cleave PGN to muramyl dipeptide (MDP), activating nucleotide-binding oligomerisation domain-containing protein 2 (NOD2) receptors in eukaryotic cells, potentially inducing anti-inflammatory host responses. Using Micrococcus luteus as a test bacterium, it was shown that muramidase from A. alcalophilum did not display antimicrobial activity, while it could cleave fluorescently labelled PGN. It was shown that the muramidase could degrade PGN down to its minimal bioactive structure MDP by using UPLC-MS/MS. Using HEK-Blue™-hNOD2 reporter cells, it was shown that the muramidase-treated PGN degradation mixture could activate NOD2. Muramidase supplementation to broiler feed increased the duodenal goblet cell and intraepithelial lymphocyte abundance while reducing duodenal wall CD3+ T lymphocyte levels. Muramidase supplementation to broiler feed only had moderate effects on the duodenal, ileal and caecal microbiome. It was shown that the newly discovered muramidase hydrolysed PGN, resulting in MDP that activates NOD2, potentially steering the host response for improved intestinal health. [ABSTRACT FROM AUTHOR]

Abstract Background In the EU conventional cages for laying hens are forbidden beginning in January 2012, however concerns about a higher transmission rate of Salmonella in alternative cages systems have been raised. The extent to which cage systems may affect the intestinal microbiota of laying hens is not known, and different microbiota may demonstrate different resistance towards colonization with Salmonella. To investigate this, ileal and caecal samples from two experimental studies where laying hens were inoculated with Salmonella Enteritidis and housed in different systems (conventional cage, furnished cage or aviary), were compared using Terminal Restriction Fragment Length Polymorphism (T-RFLP). The distribution of genera in the microbiota in caecum was furthermore described by next generation sequencing of 16S rDNA libraries. Results Hens in the same cage type developed similar T-RFLP fingerprints of the ileal and caecal microbiota, and these could be separated from layers in the other cages types. No significant difference in the fingerprint profiles could be observed between Salmonella positive and negative samples from same cage. By deep sequencing of 16S rDNA libraries from caecum, 197 different Operational Taxonomic Units (OTU) were identified, and 195 and 196 OTU respectively, were found in hens in aviary and furnished cages, but only 178 OTU of these were recovered from conventional cages. The ratio between the dominating phyla or families and genera in the microbiota remained fairly constant throughout the study. Faecalibacterium and Butyricimonas were the most prevalent genera found in the caecal microbiota of layers irrespective of the cage type. Conclusions Hens confined in the same cage group tend to develop similar microbiota in their ileum and caecum possibly due to isolation, while differences in the microbiota between cages may be caused by environmental or individual bird factors. Although the cages type had influence on composition of the microbiota in the layers by promoting higher diversity in furnished and aviary systems, we did not observe differences in colonization and excretion pattern of Salmonella from these groups. We suggest, that differences in group size and exposure to a more faecally contaminated environment in the alternative systems may explain the observed differences in diversity of the caecal microbiota.

Abstract Campylobacter contaminated broiler chicken meat is an important source of foodborne gastroenteritis and poses a serious health burden in industrialized countries. Broiler chickens are commonly regarded as a natural host for this zoonotic pathogen and infected birds carry a very high C. jejuni load in their gastrointestinal tract, especially the ceca. This eventually results in contaminated carcasses during processing. Current intervention methods fail to reduce the colonization of broiler chicks by C. jejuni due to an incomplete understanding on the interaction between C. jejuni and its avian host. Clearly, C. jejuni developed several survival and colonization mechanisms which are responsible for its highly adapted nature to the chicken host. But how these mechanisms interact with one another, leading to persistent, high-level cecal colonization remains largely obscure. A plethora of mutagenesis studies in the past few years resulted in the identification of several of the genes and proteins of C. jejuni involved in different aspects of the cellular response of this bacterium in the chicken gut. In this review, a thorough, up-to-date overview will be given of the survival mechanisms and colonization factors of C. jejuni identified to date. These factors may contribute to our understanding on how C. jejuni survival and colonization in chicks is mediated, as well as provide potential targets for effective subunit vaccine development.

Abstract Background Egg-associated transmission to humans seems to be characteristic of the Salmonella serotype Enteritidis, explaining why this particular serotype has caused a worldwide pandemic since the mid '80s. Salmonella Enteritidis is much more capable to persistently colonize the laying hen reproductive tract and to survive in the hostile egg white, as compared to other serotypes. Presentation of the hypothesis It is hypothesized that stress-induced survival mechanisms enable the serotype Enteritidis to persistently colonize the oviduct without causing damage and excessive inflammation, and to cope with the antimicrobial compounds present in egg white. Testing the hypothesis To test the hypothesis first of all Salmonella Enteritidis genes that are essential for colonization of the oviduct and survival in eggs need to be identified. Comparative genomics tools should be used to identify genes or pathogenicity islands that are present in Salmonella Enteritidis and not in the multiple non egg-contaminating serotypes. High-throughput signature-tagged-mutagenesis approaches, coupled to micro-array detection of the genes that lead to an attenuated phenotype when mutated is proposed as an ideal tool to identify genes involved in oviduct colonization and egg white survival. Identifying the stressors and antibacterial molecules in the oviduct and in the egg white that limit colonization or survival of non-Enteritidis serotypes is a second important objective that can theoretically be achieved using screenings of expressed oviduct cDNA libraries for their antibacterial activity against strains from multiple serotypes. Finally, the effect of contact with these stressors in the oviduct or egg white on Salmonella gene expression will need to be analyzed, in order to clarify whether serotype Enteritidis-specific regulation of certain stress-survival pathways are either or not present. Implications of the hypothesis Knowledge on the pathogenesis of egg infections would furthermore give insights that might be extrapolated to other biological interactions, in which a highly specialized bacterial pathogen resists the host response in a specific biological niche. In addition, this info can be of value in developing early warning criteria to identify emerging egg-associated Salmonella strains and in developing safe live attenuated vaccine strains.

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 ZnSO4.7H2O 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. [ABSTRACT FROM AUTHOR]