Dietary inclusion of a Saccharomyces cerevisiae metabolite improved reproductive performance but did not affect intestinal permeability in two chicken meat breeder lines.

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.

Nontyphoidal Salmonella infections acquired from poultry.

PURPOSE OF REVIEW: Nontyphoidal Salmonella is a major food safety concern in developed and developing countries. Table eggs are often linked to cases of foodborne gastrointestinal disease. This review is focused on the latest findings on foodborne Salmonella infections acquired from poultry products and their implications on food safety. RECENT FINDINGS: Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) are the predominant Salmonella serovars associated with human Salmonellosis. In Australia, ST is the predominant serovar but SE has been recently detected in some commercial free-range egg flocks. The Salmonella shedding in poultry flocks can be highly variable across different flocks and farms; as a result, the level of product contamination is largely attributed to the flock management. The microevolution in the ST genome after in-vivo passaging may have clinical significance. On farm use of Salmonella vaccines and/or interventions during the processing of the product can influence the bacterial load. The refrigeration of the product also influences the safety of the poultry product. SUMMARY: Many interventions are in place for the control of Salmonella from farm to fork. However, given the biosecurity challenges because of the increase in public demand for free-range products, the emergence of Salmonella virulent types and expensive diagnostics, ongoing collaborative efforts from farmers, regulators and public health officials are required.

Peroxyacetic acid and acidified sodium chlorite reduce microbial contamination on whole chicken carcasses obtained from two processing points.

Chicken meat is frequently contaminated with zoonotic bacterial pathogens such as Campylobacter spp and Salmonella spp. These two bacterial genera are commonly linked with cases of human gastrointestinal disease, thus mitigating their presence in the poultry meat supply chain is paramount. Here, the efficacy of two sanitizers, peroxyacetic acid (PAA) and acidified sodium chlorite (ASC), was tested using whole chicken carcasses obtained either prior to the inside/outside wash or the post-immersion spin chill steps of processing. Two concentrations of PAA (100 and 200 ppm) and ASC (450 and 900 ppm) were tested, and both significantly reduced total viable bacteria and Campylobacter counts per carcass. Both sanitizers also reduced the prevalence of Salmonella on whole carcasses from both processing steps. Log reduction of both the total viable and Campylobacter counts were, however, temperature and processing stage dependent. The efficacy of both PAA and ASC were also compared with sodium hypochlorite. No significant difference between the three sanitizers was observed for the reduction of TVC, Campylobacter or Salmonella using carcasses obtained at either processing step. These results demonstrate that PAA or ASC could be implemented as a replacement or used in addition to sodium hypochlorite to effectively reduce bacteria on whole carcasses.

Layers, broiler chickens and their F1 cross develop distinctly different caecal microbial communities when hatched and reared together.

AIM: To compare the caecal microbiota of layer, broiler, and intermediate F1 layer × broiler cross birds with the hypothesis that significant differences in caecal microbial composition would persist between the three groups when host and environmental interactions were minimized. METHODS AND RESULTS: Caecal contents were characterized using 16S rRNA for males of broiler (n = 12), layer (n = 12) and F1 layer × broiler cross (n = 9) birds that were hatched and reared under the same conditions. The microbial community structure differed significantly between the three groups of birds at phylum, genus and OTU levels, with clear separation of the groups observed. Firmicutes was the phylum most represented across samples; however, the high abundance of Proteobacteria in the layer birds at d28 post-hatch was unexpected, and driven by a higher abundance of E. coli. CONCLUSIONS: The microbiota phylotype between broilers, layers and their F1 cross significantly differed in community structure, diversity and relative abundance in the absence of environmental confounding, which is generally difficult to avoid in microbial studies. SIGNIFICANCE AND IMPACT OF STUDY: The results provide a unique comparison and evidence that there is a strong genetic component driving microbial composition within poultry strains, despite the embryonic development occurring in ovo.

Excreta biomarkers in response to different gut barrier dysfunction models and probiotic supplementation in broiler chickens.

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<0.001) FITC-d passage to the blood on d 21 of age, indicating a greater IP. The excreta concentrations of A1AT, I-FABP and SOD1 were unaltered by the experimental treatments. DEX increased (P<0.05) FN concentration in excreta compared with control birds. Conversely, inclusion of rye in the diet reduced (P<0.05) FN but increased (P<0.001) OVT in excreta. Independently, DEX decreased IAP (P<0.05) in excreta compared with control and rye-fed birds. The excreta concentration of LCN2 tended (P = 0.086) to increase in birds injected by DEX. There was no demonstrable effect of probiotic addition on any of the studied parameters. Among the tested biomarkers, FN, IAP, and LCN2 revealed promise as biomarkers of intestinal barrier function quantified by ELISA kits.

Nanoparticles of selenium as high bioavailable and non-toxic supplement alternatives for broiler chickens.

Selenium is commonly used in the poultry industry as an additive in broiler feed to improve immunity and overall health. The selenium comes in different forms, inorganic and organic selenium, as sodium selenite and selenomethionine, respectively. This study proposes the use of nanoparticles of selenium (nanoSe) for improved delivery and absorption of the trace element while causing no toxicity. Previous studies have shown the success in utilizing nanoSe in broiler feed, with increased absorption and diffusion of material into organs and tissues, and increased antioxidant capacity. However, the mechanism of nanoSe conversion remains unknown, and the gut microbiota is believed to play a significant role in the process. The use of inorganic selenium in poultry feed demonstrated a lower bioavailability in breast (P ≤ 0.01) and duodenum tissue (P ≤ 0.05), and increased accumulation in organs involved in detoxification processes as compared to organic selenium and selenium nanoparticle supplementation. Histopathological analysis showed that nanoSe did not cause any damaging effects to the tissues analysed, revealing intact epithelial cells in the digestive system and neuronal bodies in brain tissue. The results indicate that nanoparticles of selenium operate a similar way to organic selenium and could potentially be used in poultry feed as a trace element additive.

Oregano: A potential prophylactic treatment for the intestinal microbiota.

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.

Characterisation of the intestinal microbiota of commercially farmed saltwater crocodiles, Crocodylus porosus.

The Australian saltwater crocodile (Crocodylus porosus) industry began commercially in the 1980s, producing skins for export and crocodile meat as a by-product. Industry research has thus far focused on strategies to improve production efficiency. In the current study, we utilised 16S rRNA sequencing to characterise the intestinal microbiome of Australian saltwater crocodiles. Samples were collected from 13 commercially farmed crocodiles from six sample sites along the length of the intestinal tract. The results indicate a similar microbiome composition to that found in the freshwater alligator, with the dominate phyla represented by Firmicutes, primarily Clostridia, and Fusobacteria, which appears to be distinct from mammalian, fish, and other reptile phyla which are generally dominated by Firmicutes and Bacteroidetes. The high abundance of 'pathogenic' bacteria, with no apparent consequence to the host's health, is of great interest and warrants further additional investigation. This will enable expansion of the current understanding of host immune function and how it is modified by host and intestinal microbiome interactions.

Feed supplementation with biochar may reduce poultry pathogens, including Campylobacter hepaticus, the causative agent of Spotty Liver Disease.

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.

Correlations between intestinal innate immune genes and cecal microbiota highlight potential for probiotic development for immune modulation in poultry.

Immune function is influenced by the diversity and stability of the intestinal microbiota. A likely trade-off of immune function for growth has been demonstrated in heavier breeds of poultry that have been genetically selected for growth and feed efficiency traits. We investigated the expression of selected innate immune genes and genes encoding products involved in intestinal barrier function to determine whether function changes could be consistently linked to the phenotypic expression of feed conversion ratio (FCR), a common measure of performance within poultry broiler flocks. In addition, we compared individual cecal microbial composition with innate immune gene expression. Samples were utilised from two replicate trials termed P1E1 and P1E2. High (n = 12) and low (n = 12) performing birds were selected based on their individual FCR data from each replicate and combined for microbiota phylogenetic composition and immune gene expression analysis. Toll-like receptor 1 (TLR1La) and zonula occludens 1 (ZO1) were differentially expressed between high- and low-performing broilers. Several taxa were correlated with FCR; of these, unclassified YS2 and ZO1 were also positively correlated with each other. Interactions between taxa and differentially expressed innate immune genes between P1E1 and P1E2 were much greater compared to relationships between high- and low-performing birds. At the level of phylum, reciprocal correlations between tight junction proteins and Toll-like receptors with Bacteroidetes and Firmicutes were evident, as were correlations at the genus level.

Transcriptional analysis of liver from chickens with fast (meat bird), moderate (F1 layer x meat bird cross) and low (layer bird) growth potential.

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 < 0.05, and a false discovery rate of < 0.05, with any fold change considered. In total, 6278 genes were found to be DE with 5832 DE between meat birds and layers (19%), 2935 DE between meat birds and the cross (9.6%) and 493 DE between the cross and layers (1.6%). Comparisons between the three groups identified 155 significant DE genes. Gene ontology (GO) enrichment and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis of the 155 DE genes showed the FoxO signalling pathway was most enriched (P = 0.001), including genes related to cell cycle regulation and insulin signalling. Significant GO terms included 'positive regulation of glucose import' and 'cellular response to oxidative stress', which is also consistent with FoxOs regulation of glucose metabolism. There were high correlations between FoxO pathway genes and bodyweight, as well as genes related to glycolysis and bodyweight. CONCLUSIONS: This study revealed large transcriptome differences between meat and layer birds. There was significant evidence implicating the FoxO signalling pathway (via cell cycle regulation and altered metabolism) as an active driver of growth variations in chicken. Functional analysis of the FoxO genes is required to understand how they regulate growth and egg production.

Evaluation of fatty acid metabolism and innate immunity interactions between commercial broiler, F1 layer × broiler cross and commercial layer strains selected for different growth potentials.

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 ß-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.