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One of the commonly used methods to control on-farm Salmonella outbreaks in the poultry industry is vaccination. Vaccine diluents may affect vaccine efficacy but there are limited studies on how different diluents affect the colonization of live, attenuated vaccines. In this study, buffered peptone water, Marek's diluent and water were used as the diluents to reconstitute the commercially available Salmonella Typhimurium vaccine (Vaxsafe® ST). The diluent reconstituted vaccine was then given to day-old layer hens to test its effects on gut colonization. The vaccine reconstituted in Marek's diluent and buffered peptone water had a significantly higher colonization rate at 7 days post-vaccination in liver, spleen, oesophagus, crop, and proventriculus. At 14 days post-vaccination, there was a significantly higher vaccine load in the upper digestive tract of chickens that received vaccine reconstituted in Marek's diluent and buffered peptone water. The Marek's diluent and buffered peptone water reconstituted vaccine had a higher detection rate in the chicken's liver and spleen. Colonisation with water as a diluent was higher than the others especially in the lower parts of the intestine at 7 days post-vaccination. There was non-significant variation in the antibody titers at 7- and 14-days post-vaccination. In general, the results demonstrated that the use of nutrient-rich diluents in reconstituting live, attenuated Salmonella vaccine can enhance vaccine colonization. This study establishes a foundation for future work aimed at optimising strategies for vaccine reconstitution, crucial for eliciting a robust immune response in the host.

ABSTRACT: Gastrointestinal dysbiosis is a disturbance in mucosal homeostasis, producing low-grade chronic intestinal inflammation and impaired intestinal barrier function. It is induced by several factors, including nutrition and stress, which are both significant factors when considering current broiler breeder practices. A great grandparent (GGP) chicken meat line was identified displaying clinical signs characteristic of potential dysbiosis, including wet droppings and litter, in addition to reduced reproductive performance when compared to a consistently high performing line. This study aimed to determine whether the reduced reproductive performance observed in these hens was a result of dysbiosis and whether dietary supplementation with a Saccharomyces cerevisiae (SC) fermentation product would alleviate clinical signs. Dietary inclusion of SC did not influence intestinal permeability, WBC differentials, or corticosterone concentration in either the wet litter (WL) or high-performing (HP) breeder lines. Compared to hens from the HP line, WL line hens had a significant increase in intestinal permeability at 26 wk (onset of lay). WL hen heterophil counts were increased markedly at week 26 before declining. At weeks 26, 32, and 37 there were also significant increases in monocytes. Higher plasma corticosterone was also observed in WL hens at 37 wk. No significant differences in heterophil to lymphocyte (H:L) ratios or feather corticosterone were observed between lines. Dietary inclusion of SC supplementation to breeder diets had some benefit in regards to reducing hen mortality, improving egg production and hatchability but only in the WL line. Results from this study did not indicate that hens from the wet litter line were experiencing gut dysbiosis. Chronic intestinal inflammation may be a possible reason for the increase in intestinal permeability. These results do indicate that both breeder lines may be exhibiting physiological stress. Future investigation into the physiology and behavior around point of lay is required to find novel strategies to alleviate this stress and in turn, potentially improve welfare and production outcomes.

Abstract Background Divergent selection for meat and egg production in poultry has resulted in strains of birds differing widely in traits related to these products. Modern strains of meat birds can reach live weights of 2 kg in 35 d, while layer strains are now capable of producing more than 300 eggs per annum but grow slowly. In this study, RNA-Seq was used to investigate hepatic gene expression between three groups of birds with large differences in growth potential; meat bird, layer strain as well as an F1 layer x meat bird. The objective was to identify differentially expressed (DE) genes between all three strains to elucidate biological factors underpinning variations in growth performance. Results RNA-Seq analysis was carried out on total RNA extracted from the liver of meat bird (n = 6), F1 layer x meat bird cross (n = 6) and layer strain (n = 6), males. Differential expression of genes were considered significant at P

Abstract Background The broiler industry has undergone intense genetic selection over the past 50 yr. resulting in improvements for growth and feed efficiency, however, significant variation remains for performance and growth traits. Production improvements have been coupled with unfavourable metabolic consequences, including immunological trade-offs for growth, and excess fat deposition. To determine whether interactions between fatty acid (FA) metabolism and innate immunity may be associated with performance variations commonly seen within commercial broiler flocks, total carcass lipid %, carcass and blood FA composition, as well as genes involved with FA metabolism, immunity and cellular stress were investigated in male birds of a broiler strain, layer strain and F1 layer × broiler cross at d 14 post hatch. Heterophil: lymphocyte ratios, relative organ weights and bodyweight data were also compared. Results Broiler bodyweight (n = 12) was four times that of layers (n = 12) by d 14 and had significantly higher carcass fat percentage compared to the cross (n = 6; P = 0.002) and layers (P = 0.017) which were not significantly different from each other (P = 0.523). The carcass and whole blood FA analysis revealed differences in the FA composition between the three groups indicating altered FA metabolism, despite all being raised on the same diet. Genes associated with FA synthesis and β-oxidation were upregulated in the broilers compared to the layers indicating a net overall increase in FA metabolism, which may be driven by the larger relative liver size as a percentage of bodyweight in the broilers. Genes involved in innate immunity such as TLR2 and TLR4, as well as organelle stress indicators ERN1 and XBP1 were found to be non-significant, with the exception of high expression levels of XBP1 in layers compared to the cross and broilers. Additionally there was no difference in heterophil: lymphocytes between any of the birds. Conclusions The results provide evidence that genetic selection may be associated with altered metabolic processes between broilers, layers and their F1 cross. Whilst there is no evidence of interactions between FA metabolism, innate immunity or cellular stress, further investigations at later time points as growth and fat deposition increase would provide useful information as to the effects of divergent selection on key metabolic and immunological processes.

Increased intestinal permeability (IP) and inflammation are both linked with functionality of the intestinal barrier and in particular enterocytes. Currently, almost all assessment methods of the intestinal barrier function are invasive. The present study aimed to quantify selected proteins as novel biomarkers in excreta of broiler chickens to facilitate non-invasive assessment of gut barrier function using enzyme-linked immunosorbent assays (ELISA). It was further hypothesised that probiotics as feed additives may counteract gut barrier dysfunction. A 3 × 2 factorial arrangement of treatments was used with the main factors being gut barrier dysfunction models (control, rye-based diet, and dexamethasone-DEX) with and without probiotic supplementation (a three-strain Bacillus) using 72 male Ross 308 day-old chickens. Each of the 6 experimental treatments was replicated 12 times. On d 21 of age, fluorescein isothiocyanate dextran (FITC-d) uptake into serum was examined to test IP. Fresh excreta samples were collected on d 20. The biomarkers included alpha-1 antitrypsin (A1AT), intestinal fatty acid binding protein (I-FABP), lipocalin-2 (LCN2), fibronectin (FN), intestinal alkaline phosphatase (IAP), ovotransferrin (OVT) and superoxide dismutase [Cu-Zn] (SOD1). Only DEX increased (P

Prophylactic use of antibiotics in poultry diets has been identified as a problematic practice because of its potential to exacerbate the spread of antibiotic resistance to human pathogens. A range of countries have opted to completely ban the use of antibiotics in animal feed. The animal production industries are looking for alternative ways to effectively control pathogens while providing the performance benefits previously secured by antibiotics in feed. Here, we present evidence that oregano (Origanum vulgare) could be a potential alternative for pathogen control in the poultry industry. Broiler diets were supplemented with oregano powder (0%, 0.5%, 1%, and 2%) for six weeks. The capacity for pathogen control was estimated by microbiota profiling of the jejunum, ileum, and caecum content, and in the faeces, by 16S rRNA gene amplicon sequencing. The concentrations of short-chain fatty acids in the caecal content were also measured, as were villus/crypt parameters in the ileum. There were no differences among treatments in weight gain, feed intake, or the concentration of short-chain fatty acids. The height, width, and the surface area of villi in the ileum were not influenced by oregano addition. However, 1% and 2% of oregano produced a significant increase in the villus height to crypt depth ratio. There were no visible histopathological changes in the liver in control and treated groups. Although oregano had no significant effect on overall microbial diversity and gross composition, some specific genera, like Proteus, Klebsiella and Staphylococcus, which include known pathogens, were reduced in relative abundance by oregano treatment. Bifidobacterium, recognized as a beneficial and probiotic genus, was also suppressed by the oregano treatment.

This study evaluated the impact of vaccine diluents (peptone or water) on the protective effects of Salmonella Typhimurium (S. Typhimurium) vaccine. Vaccinated broilers were challenged with different doses of wild-type S. Typhimurium through dust. At the time of cull, vaccine load was highest in caeca and lowest in spleen. Wild-type S. Typhimurium was detectable after 24 hrs only in the vaccinated birds challenged with 108 CFU and positive control. S. Typhimurium load was lower in the organs of the groups challenged with 104 and 106 compared to the 108 CFU group. The caecal microbiota alpha diversity of the vaccinated or vaccinated and challenged chickens differed from the positive and negative control groups. Beta diversity of the positive control clustered separately from all other treatment groups, showing that vaccine caused minimal changes in gut microbiota structure. The vaccinated and/or wild-type challenged chickens showed significantly higher abundance of Anaerostignum, Lachnoclostridium, Intestinimonas, Colidextribacter, Monoglobus, Acetanaerobacterium and Subdoligranulum. Outcomes from this study demonstrate that the vaccine effectively protected broiler chickens from S. Typhimurium infection and helped maintain a more stable gut microbiota structure, reducing the impact of S. Typhimurium on gut health. Vaccine diluent did not affect gut microbiota composition.

Increased global regulation and restrictions on the non-therapeutic use of antibiotics in the poultry industry means that there is a need to identify alternatives that prevent infection while still conveying the growth and performance benefits afforded by their use. Biochars are produced by the incomplete pyrolysis of organic materials, with reports of use as a feed supplement and activity against pathogenic bacteria. In the current study the dose-dependent effects of biochar dietary inclusion in layer diets at 1%, 2% and 4% w/w were investigated to determine a) the efficacy of biochar as an anti-pathogenic additive on the intestinal microbiota and b) the optimal inclusion level. Biochar inclusion for anti-pathogenic effects was found to be most beneficial at 2% w/w. Poultry pathogens such as Gallibacterium anatis and campylobacters, including Campylobacter hepaticus, were found to be significantly lower in biochar fed birds. A shift in microbiota was also associated with the incorporation of 2% w/w biochar in the feed in two large scale trials on two commercial layer farms. Biochar inclusion for anti-pathogenic effects was found to be most beneficial at 2% w/w. Differential effects of the timing of biochar administration (supplementation beginning at hatch or at point of lay) were also evident, with greater impact on community microbial structure at 48 weeks of age when birds were fed from hatch rather than supplemented at point of lay.