Nutritional Supplement of Hatchery Eggshell Membrane Improves Poultry Performance and Provides Resistance against Endotoxin Stress.
ABSTRACT: Eggshells are significant part of hatchery waste which consist of calcium carbonate crust, membranes, and proteins and peptides of embryonic origins along with other entrapped contaminants including microbes. We hypothesized that using this product as a nutritional additive in poultry diet may confer better immunity to the chickens in the paradigm of mammalian milk that enhances immunity. Therefore, we investigated the effect of hatchery eggshell membranes (HESM) as a short term feed supplement on growth performance and immunity of chickens under bacterial lipopolysaccharide (LPS) challenged condition. Three studies were conducted to find the effect of HESM supplement on post hatch chickens. In the first study, the chickens were fed either a control diet or diets containing 0.5% whey protein or HESM as supplement and evaluated at 5 weeks of age using growth, hematology, clinical chemistry, plasma immunoglobulins, and corticosterone as variables. The second and third studies were done to compare the effects of LPS on control and HESM fed birds at 5 weeks of age following at 4 and 24 h of treatment where the HESM was also sterilized with ethanol to deplete bacterial factors. HESM supplement caused weight gain in 2 experiments and decreased blood corticosterone concentrations. While LPS caused a significant loss in body weight at 24 h following its administration, the HESM supplemented birds showed significantly less body weight loss compared with the control fed birds. The WBC, heterophil/lymphocyte ratio, and the levels of IgG were low in chickens fed diets with HESM supplement compared with control diet group. LPS challenge increased the expression of pro-inflammatory cytokine gene IL-6 but the HESM fed birds showed its effect curtailed, also, which also, favored the up-regulation of anti-inflammatory genes compared with control diet fed chickens. Post hatch supplementation of HESM appears to improve performance, modulate immunity, and increase resistance of chickens to endotoxin.
Project description:A total of 360 male Ross 308 broiler chickens were used in a feeding study to assess the influence of macronutrients and energy density on feed intakes from 10 to 31 days post-hatch. The study comprised ten dietary treatments from five dietary combinations and two feeding approaches: sequential and choice feeding. The study included eight experimental diets and each dietary combination was made from three experimental diets. Choice fed birds selected between three diets in separate feed trays at the same time; whereas the three diets were offered to sequentially fed birds on an alternate basis during the experimental period. There were no differences between starch and protein intakes between choice and sequentially fed birds (P > 0.05) when broiler chickens selected between diets with different starch, protein and lipid concentrations. When broiler chickens selected between diets with different starch and protein but similar lipid concentrations, both sequentially and choice fed birds selected similar ratios of starch and protein intake (P > 0.05). However, when broiler chickens selected from diets with different protein and lipid but similar starch concentrations, choice fed birds had higher lipid intake (129 versus 118 g/bird, P = 0.027) and selected diets with lower protein concentrations (258 versus 281 g/kg, P = 0.042) than birds offered sequential diet options. Choice fed birds had greater intakes of the high energy diet (1471 g/bird, P < 0.0001) than low energy (197 g/bird) or medium energy diets (663 g/bird) whilst broiler chickens were offered diets with different energy densities but high crude protein (300 g/kg) or digestible lysine (17.5 g/kg) concentrations. Choice fed birds had lower FCR (1.217 versus 1.327 g/g, P < 0.0001) and higher carcass yield (88.1 versus 87.3%, P = 0.012) than sequentially fed birds. This suggests that the dietary balance between protein and energy is essential for optimal feed conversion efficiency. The intake path of macronutrients from 10-31 days in choice and sequential feeding groups were plotted and compared with the null path if broiler chickens selected equal amounts of the three diets in the combination. Regardless of feeding regimen, the intake paths of starch and protein are very close to the null path; however, lipid and protein intake paths in choice fed birds are father from the null path than sequentially fed birds.
Project description:BACKGROUND:Fatness is an unwanted side effect of genetic selection in broiler chickens. In this study, we introduce mountain celery powder as a feed supplement to suppress lipogenesis and improve antioxidant status in broiler chickens. Male broiler chicks (Ross 308) were fed a control diet or a diet that includes mountain celery (MC) at 7.5 gkg-1over 42 days. RESULTS:Body weight gain and feed conversion ratio significantly (P < 0.05) improved in chicks fed MC. A highly significant down-regulation of genes involved in hepatic lipogenesis including acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), malic enzyme (ME), and lipoprotein lipase (LPL) was observed in the liver of chickens fed MC. These birds, however, had greater compensatory upregulation in antioxidative genes SOD1 and catalase in the liver compared to the birds that received the control diet. Birds received MC had significantly lower level of lipid peroxidation (1.59 μmol/L serum malondialdehyde) compared to birds from the control group (3.57 μmol/L; P = 0.0024). Birds fed MC had significantly (P < 0.05) lower circulatory concentrations of triacylglycerols, cholesterol, and LDL but higher concentrations of HDL. Relative liver weight and abdominal fat deposition were significantly reduced by feeding MC. CONCLUSIONS:It can be concluded that feeding birds MC significantly suppresses hepatic lipogenesis by down-regulating key hepatic lipogenic enzyme genes and boosts antioxidant capacity by up-regulating hepatic antioxidantive genes.
Project description:BACKGROUND:The present study was conducted to investigate the effects of dietary plant-derived phytonutrients, carvacrol, cinnamaldehyde and Capsicum oleoresin, on the translational regulation of genes associated with immunology, physiology and metabolism using high-throughput microarray analysis and in vivo disease challenge model of avian coccidiosis. METHODS:In this study, we used nutrigenomics technology to investigate the molecular and genetic mechanisms of dietary modulation of host innate immunity and metabolism by three phytonutrients. To validate their immunomodulatory effects in a disease model, young broiler chickens fed a standard diet supplemented with three phytochemicals (carvacrol, cinnamaldehyde, and Capsicum oleoresin) from one day post-hatch were orally challenged with E. acervulina. The body weight gain and fecal oocyst production were used to evaluate coccidiosis disease parameters. RESULTS:Analysis of global gene expression profiles of intestinal tissues from phytonutrient-fed birds indicated that Capsicum oleoresin induced the most gene changes compared to the control group where many of these genes were associated with those of metabolism and immunity. The most reliable network induced by dietary cinnamaldehyde treatment was related with the functions of antigen presentation, humoral immune response, and inflammatory disease. Furthermore, dietary supplementation with these phytonutrients significantly protected broiler chickens against live coccidiosis challenge infection based on body weight and parasite fecundity. CONCLUSIONS:The results of this study provide clear evidence to support the idea that plant-derived phytochemicals possess immune-enhancing properties in chickens and these new findings create a new possibility to develop effective drug-free alternative strategies for disease control for poultry infectious diseases.
Project description:The gut not only plays a key role in digestion and absorption of nutrients but also forms a physical barrier and first line of defense between the host and the luminal environment. A functional gut barrier (mucus and epithelial cells with tight junctions [TJ]) is essential for optimal health and efficient production in poultry. In current broiler system, chicks are deprived of food and water up to 72 h due to uneven hatching, hatchery procedures, and transportation. Post-hatch feed delay results in lower BW, higher FCR and mortality, and delayed post-hatch gut development. Little is known about the effects of early neonatal development and delayed feeding immediately post-hatch on gut barrier function in chickens. Therefore, the aim of the present study was to characterize the expression pattern of gut barrier-related and TJ-related genes in the small intestine of broiler chickens during early development and delay in access to feed. Newly hatched chicks received feed and water immediately after hatch or were subjected to 48 h delayed access to feed to mimic commercial hatchery setting and operations. Birds were sampled (n = 6) at -48, 0, 4, 24, 48, 72, 96, 144, 192, 240, 288, and 336 h post-hatch. Jejunum and ileum were collected, cleaned of digesta, and snap-frozen in liquid nitrogen or fixed in paraformaldehyde. The relative mRNA levels of gut barrier- and TJ-related protein genes were measured by quantitative PCR and analyzed by 2-way ANOVA. In both tissues, changes (P < 0.05) in gene expression pattern of gut barrier-related and TJ-related genes were detected due to delayed access to feed post-hatch and/or development. In general, expression of TJ-related genes was downregulated while mRNA levels of gut barrier-related genes were upregulated during development. Histological differences and changes in mucin staining due to age and treatment were observed. These results suggest that delayed access to feed post-hatch may affect TJ structure and/or function and therefore gut barrier function and overall health of the chicken small intestine.
Project description:Subtherapeutic levels of dietary antibiotics increase growth performance in domestic animals, but the mechanisms are poorly understood. Here, 1-week-old broiler chickens were challenged with LPS (experiment 1), or co-infected with Eimeria maxima and Clostridium perfringens as an experimental model of necrotic enteritis (experiment 2), and fed a standard basal diet or a basal diet supplemented with virginiamycin or bacitracin methylene disalicylate. In experiment 1, LPS-challenged chickens fed the unsupplemented diet had decreased body weight gains, compared with unsupplemented controls given the PBS control. In contrast, antibiotic supplementation increased body weight gains in both the LPS-challenged and PBS groups, compared with the antibiotic-free diet. LPS-challenged chickens fed the unsupplemented diet had increased expression levels of intestinal tight junction proteins (ZO1, JAM2), MUC2 gel-forming mucin, and inflammatory cytokines (IL-1?, IL-2, IL-6, IL-8, IL-17A) at 24 h post-challenge, compared with unsupplemented chickens given the PBS control. However, LPS-challenged chickens fed the antibiotic-supplemented diets had decreased levels of intestinal inflammatory cytokine transcripts, compared with LPS-challenged chickens given the unsupplemented basal diet. In experiment 2, E. maxima/C. perfringens-co-infected chickens fed the antibiotic-supplemented diets had increased body weight gains, decreased intestinal pathology, and greater intestinal crypt depth, compared with co-infected chickens given the unsupplemented diet. Further, similar to LPS challenge, E. maxima/C. perfringens-co-infection of chickens fed the antibiotic-supplemented diets decreased expression levels of intestinal inflammatory cytokines, compared with co-infected chickens given the unsupplemented diet. These results support the hypothesis that dietary antibiotic growth promoters might increase poultry growth, in part, through down-regulation of pathogen-induced inflammatory responses.
Project description:Salmonella causes inflammation in infected hosts. Inflammation is a well-characterized defensive mechanism of innate immunity. The recognition and engagement of lipopolysaccharide (LPS) endotoxins in the outer membranes of Salmonella to Toll-like receptor 4 of immune cells (macrophages and dendritic cells) trigger inflammatory responses characterized by secretion of pro-inflammatory cytokines, including TNF-beta, IL-1 and IL-6. These cytokines cause fever, anorexia, bodyweight losses, and catabolism of skeletal muscles and adipose tissues. However, molecular events underlying innate immune responses and metabolic activities during the later stage of inflammation are poorly understood. Additionally, the effects of prebiotics and antibiotics on innate immunity and nutrient metabolism are not yet reported. The objective of this study is to investigate the effects of a mannanoligosaccharide (MOS) prebiotic and virginiamycin (VIRG) sub-therapeutic antibiotic on innate immunity and glucose metabolism during late inflammation. We induced Salmonella LPS-systemic inflammation in a chicken model. Differentially regulated gene expressions were measured using 2 colour focussed oligonucleotide chicken-specific microarrays. Microarray analysis was performed on liver, intestinal and skeletal muscle tissues. We found that late inflammation was principally modulated by interleukin 3 (IL 3) and that glucose was mobilized from gluconeogenesis occurring in the intestines only. MOS and VIRG modulated innate immunity and metabolic genes differently. In contrast to VIRG, MOS terminated inflammatory responses earlier. Our results indicate IL 3 gene up-regulation in VIRG-fed chickens. To meet the higher energy requirements of VIRG chickens, genes for intestinal gluconeogenesis and liver glycolysis were respectively induced. Our study reveals the potential mechanisms by which prebiotic and antibiotic modulated innate immunity and glucose metabolism during late inflammation. 14-day old chickens were injected i.p. with saline or LPS. For each tissue and experimental conditions (saline or LPS challenge), a total of 12 microarrays (6 MOS birds + 6 VIRG birds) were used in a 2 x 2 factorial design and complete interwoven loop arrangement. We compared gene expression from prebiotic-fed birds with antibiotic-fed birds without including reference RNA. LPS challenge, antibiotic or prebiotic, innate immunity, glucose metabolism
Project description:Diminishing the cost of broiler chicken diet is a critical issue in the poultry industry. Numerous studies were performed to achieve this pivotal objective by diet supplementation with alternative feed additives. In the current study, low-energy broiler rations were supplemented with different commercial multienzyme formulations to minimize the cost, and increase the digestibility and absorption of the digested macronutrients. Cobb Avian 48 broiler chicks (mixed sex, 1-d-old, n = 3120) were randomly allocated into six groups, and each group was subdivided into four replicates (130 birds per replicate). The birds were randomly allocated into a control group fed basal diet (CB); control group fed low-energy diet (CL); and birds fed low-energy diets supplemented with different enzyme formulations. The enzyme formulations used were Xylam 500® (CLX group), Hemicell® (CLH group), Avizyme® (CLA group), and Megazyme® (CLM group,) following the doses recommended by the manufacturers. The growth performance of CLA and CLH group birds was significantly improved when compared with CL. In comparison with CB, Avizyme® significantly (p < 0.001) increased the intestinal PEPT1, GLUT2, ACC, and IL-2 expression; PEPT1 facilitates the absorption of micronutrients. In conclusion, exogenous multienzyme complexes may be included in the low-energy diet to enhance the performance of broiler chickens (Avizyme® ? Hemicell® ? Megazyme®), and reduce the diet cost by up-regulating the expression of intestinal nutrient transporter genes, and improving the immunity and serum biochemical parameters of broiler chickens.
Project description:The identities of genes that underlie population variation in adipose tissue development in farm animals are poorly understood. Previous studies in our laboratory have suggested that increased fat tissue involves the expression modulation of an array of genes in broiler chickens. Of special interest are eight genes, FGFR3, EPHB2, IGFBP2, GREM1, TNC, COL3A1, ACBD7, and SCD. To understand their expression regulation and response to dietary manipulation, we investigated their mRNA levels after dietary manipulation during early development. Chickens were fed either a recommended standard or a high caloric diet from hatch to eight weeks of age (WOA). The high caloric diet markedly affected bodyweight of the broiler birds. mRNA levels of the eight genes in the abdominal adipose tissue were assayed at 2, 4, 6, and 8 WOA using RT-qPCR. Results indicate that (1) FGFR3 mRNA level was affected significantly by diet, age, and diet:age interaction; (2) COL3A mRNA level was repressed by high caloric diet; (3) mRNA levels of EPHB2, ACBD7, and SCD were affected by age; (4) mRNA level of TNC was modulated by age:diet interaction; (5) changes in GREM1 and IGFBP2 mRNA levels were not statistically different.
Project description:This study was conducted to investigate the effect of carotenoid, oligosaccharide and anthocyanin supplementation in broiler diets under Escherichia coli lipopolysaccharide (LPS) challenge. Ross 308 chickens were fed 5 diets: basal diet (control diet), diet supplemented with ?-glucan in 0.05% (positive control) and diets with 0.5% carotenoid-, oligosaccharide- or anthocyanin contents. On the 26th days of age, chickens were challenged intraperitoneally 2 mg LPS per kg of body weight. 12 h after injection, birds were euthanized, then spleen and ileum samples were collected. LPS induced increased relative mRNA expression of splenic (p = 0.0445) and ileal (p = 0.0435) interleukin-1? (IL-1?), which was lower in the spleen in carotenoid (p = 0.0114), oligosaccharide (p = 0.0497) and anthocyanin (p = 0.0303)-treated chickens compared to LPS-injected control birds. Dietary supplementation of carotenoids also decreased relative gene expression of splenic interleukin-6 (IL-6) (p = 0.0325). In the ileum, ?-glucan supplementation showed lower relative mRNA expression of toll-like receptor 5 (TLR-5) (p = 0.0387) compared to anthocyanin treatment. Gene expression of both splenic and ileal interferon-? (IFN-?), interferon-? (IFN-?), toll-like receptor 4 (TLR-4) and toll-like receptor 5 (TLR-5) were not influenced by dietary supplements. In conclusion, carotenoids, oligosaccharides and anthocyanins could partially mitigate the immune stress caused by LPS challenge. All of the compounds impacted longer villus height (p < 0.0001), villus height:crypt depth ratios were higher after ?-glucan (p < 0.0001) and anthocyanin (p = 0.0063) supplementations and thickened mucosa was observed in ?-glucan (p < 0.0001), oligosaccharide (p < 0.0001) and anthocyanin (p = 0.048) treatments. All of these findings could represent a more effective absorption of nutrients.
Project description:The mammalian hippocampus is particularly vulnerable to chronic stress. Adult neurogenesis in the dentate gyrus is suppressed by chronic stress and by administration of glucocorticoid hormones. Post-natal and adult neurogenesis are present in the avian hippocampal formation as well, but much less is known about its sensitivity to chronic stressors. In this study, we investigate this question in a commercial bird model: the broiler breeder chicken. Commercial broiler breeders are food restricted during development to manipulate their growth curve and to avoid negative health outcomes, including obesity and poor reproductive performance. Beyond knowing that these chickens are healthier than fully-fed birds and that they have a high motivation to eat, little is known about how food restriction impacts the animals' physiology. Chickens were kept on a commercial food-restricted diet during the first 12 weeks of life, or released from this restriction by feeding them ad libitum from weeks 7-12 of life. To test the hypothesis that chronic food restriction decreases the production of new neurons (neurogenesis) in the hippocampal formation, the cell proliferation marker bromodeoxyuridine was injected one week prior to tissue collection. Corticosterone levels in blood plasma were elevated during food restriction, even though molecular markers of hypothalamic-pituitary-adrenal axis activation did not differ between the treatments. The density of new hippocampal neurons was significantly reduced in the food-restricted condition, as compared to chickens fed ad libitum, similar to findings in rats at a similar developmental stage. Food restriction did not affect hippocampal volume or the total number of neurons. These findings indicate that in birds, like in mammals, reduction in hippocampal neurogenesis is associated with chronically elevated corticosterone levels, and therefore potentially with chronic stress in general. This finding is consistent with the hypothesis that the response to stressors in the avian hippocampal formation is homologous to that of the mammalian hippocampus.