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The mechanism of sex determination in chickens, especially the molecular mechanism of female ovarian development, has not yet been fully elucidated. Previous studies have shown that RSPO1, which is associated with ovarian development in mammals, might have a conserved role in chickens. In this study, we systematically investigated the spatiotemporal expression pattern of RSPO1 in various tissues, especially gonads, of male and female chicken embryos using qPCR and Western blotting, and we explored its correlation with the expression of key genes in the estrogen pathway using drug treatment or gene overexpression in vivo and in vitro. Our results reveal that RSPO1 was widely expressed in all examined tissues of chicken embryos, showing a female bias in gonadal tissues at both the mRNA and protein levels. Surprisingly, RSPO1 was not differentially expressed between male and female gonadal cells with fadrozole-induced estrogen pathway blockades, and furthermore, estradiol-induced estrogen stimulation altered the expression of RSPO1. In addition, overexpression of RSPO1 in gonadal cells induced the mRNA expression of its downstream target genes, Wnt family member 4 (WNT4) and Catenin beta 1 (CTNNB1), and that of estrogen receptor α (ERα), an estrogen pathway gene. In summary, this study provided new evidence for elucidating the role of RSPO1 in ovarian development in poultry.

The sex chromosomes of birds are designated Z and W. The male is homogamous (ZZ), and the female is heterogamous (ZW). The chicken W chromosome is a degenerate version of the Z chromosome and harbors only 28 protein-coding genes. We studied the expression pattern of the W chromosome gene MIER3 (showing differential expression during gonadogenesis) in chicken embryonic gonads and its potential role in gonadal development. The W copy of MIER3 (MIER3–W) shows a gonad-biased expression in chicken embryonic tissues which was different from its Z copy. The overall expression of MIER3–W and MIER3–Z mRNA and protein is correlated with the gonadal phenotype being higher in female gonads than in male gonads or female-to-male sex-reversed gonads. Chicken MIER3 protein is highly expressed in the nucleus, with relatively lower expression in the cytoplasm. Overexpression of MIER3–W in male gonad cells suggested its effect on the GnRH signaling pathway, cell proliferation, and cell apoptosis. MIER3 expression is associated with the gonadal phenotype. MIER3 may promote female gonadal development by regulating EGR1 and αGSU genes. These findings enrich our knowledge of chicken W chromosome genes and support a more systematic and in-depth understanding of gonadal development in chickens.

Nutrition and energy levels have an important impact on animal growth, production performance, disease occurrence and health recovery. Previous studies indicate that melanocortin 5 receptor (MC5R) is mainly involved in the regulations of exocrine gland function, lipid metabolism and immune response in animals. However, it is not clear how MC5R participates in the nutrition and energy metabolism of animals. To address this, the widely used animal models, including the overfeeding model and the fasting/refeeding model, could provide an effective tool. In this study, the expression of MC5R in goose liver was first determined in these models. Goose primary hepatocytes were then treated with nutrition/energy metabolism-related factors (glucose, oleic acid and thyroxine), which is followed by determination of MC5R gene expression. Moreover, MC5R was overexpressed in goose primary hepatocytes, followed by identification of differentially expressed genes (DEGs) and pathways subjected to MC5R regulation by transcriptome analysis. At last, some of the genes potentially regulated by MC5R were also identified in the in vivo and in vitro models, and were used to predict possible regulatory networks with PPI (protein–protein interaction networks) program. The data showed that both overfeeding and refeeding inhibited the expression of MC5R in goose liver, while fasting induced the expression of MC5R. Glucose and oleic acid could induce the expression of MC5R in goose primary hepatocytes, whereas thyroxine could inhibit it. The overexpression of MC5R significantly affected the expression of 1381 genes, and the pathways enriched with the DEGs mainly include oxidative phosphorylation, focal adhesion, ECM–receptor interaction, glutathione metabolism and MAPK signaling pathway. Interestingly, some pathways are related to glycolipid metabolism, including oxidative phosphorylation, pyruvate metabolism, citrate cycle, etc. Using the in vivo and in vitro models, it was demonstrated that the expression of some DEGs, including ACSL1, PSPH, HMGCS1, CPT1A, PACSIN2, IGFBP3, NMRK1, GYS2, ECI2, NDRG1, CDK9, FBXO25, SLC25A25, USP25 and AHCY, was associated with the expression of MC5R, suggesting these genes may mediate the biological role of MC5R in these models. In addition, PPI analysis suggests that the selected downstream genes, including GYS2, ECI2, PSPH, CPT1A, ACSL1, HMGCS1, USP25 and NDRG1, participate in the protein–protein interaction network regulated by MC5R. In conclusion, MC5R may mediate the biological effects caused by changes in nutrition and energy levels in goose hepatocytes through multiple pathways, including glycolipid-metabolism-related pathways.

The development of mammalian nonalcoholic fatty liver disease is associated with oxidative stress, reduced mitochondrial function, and increased apoptosis in hepatocytes; however, the expressions of mitochondria-related genes are elevated in goose fatty liver, suggesting that there may be a unique protective mechanism in goose fatty liver. The aim of the study was to investigate this protective mechanism in terms of anti-oxidant capacity. Our data showed no substantial differences in the mRNA expression levels of the apoptosis-related genes including B-cell lymphoma-2 (Bcl-2), BCL2-associated X (Bax), cysteinyl aspartate-specific proteinase-3 (Caspase-3), and cysteinyl aspartate-specific proteinase-9 (Caspase-9) in the livers of the control and overfeeding Lander geese groups. The protein expression levels of Caspase-3 and cleaved Caspase-9 were not markedly different between the groups. Compared with the control group, malondialdehyde content was significantly lower (P < 0.01), glutathione peroxidase (GSH-Px) activity, glutathione (GSH) content, and mitochondrial membrane potential levels were higher (P < 0.01) in the overfeeding group. The mRNA expression levels of the anti-oxidant genes superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPX1), and glutathione peroxidase 2 (GPX2) were increased in goose primary hepatocytes after 40 mM and 60 mM glucose treatment. Reactive oxygen species (ROS) levels were significantly reduced (P < 0.01), whereas the mitochondrial membrane potential was maintained at normal levels. The mRNA expression levels of the apoptosis-related genes Bcl-2, Bax, and Caspase-3 were not substantial. There were no significant differences in the expression levels of Caspase-3 and cleaved Caspase-9 proteins. In conclusion, glucose-induced enhanced anti-oxidant capacity may help protect the function of mitochondria and inhibit the occurrence of apoptosis in goose fatty liver.

Background The sex chromosomes of birds are designated Z and W. The male is homogamous (ZZ), and the female is heterogamous (ZW). The chicken W chromosome is a degenerate version of the Z chromosome and harbors only 28 protein-coding genes. We studied the expression pattern of the W chromosome gene MIER3 (showing differential expression during gonadogenesis) in chicken embryonic gonads and its potential role in gonadal development. Results The W copy of MIER3 (MIER3-W) showed a gonad-baised expression in chicken embryonic tissues which was different with its Z copy. The conbined expression of MIER3 mRNA and protein were correlated with the gonadal phenotype, being higher in female gonads than in male gonads or female-to-male sex reversed gonads. Chicken MIER3 protein was highly expressed in the nucleus, with relatively lower expression in the cytoplasm. Overexpression of MIER3-W in male gonad cells suggested its effcet on GnRH signaling pathway (potentially through EGR1 and αGSU), cell proliferation and cell apoptosis. However, at the in vivo level, MIER3 overexpression did not change the sex phenotype of chicken embryos. Conclusions MIER3 expression was associated with the gonadal phenotype. It affected multiple signaling pathways and inhibited cell apoptosis. MIER3 may promote female gonadal development by regulating EGR1 and αGSU genes; however, it does not determine the sex differentiation of gonads. These findings enrich our knowledge of chicken W chromosome genes and support a more systematic and in-depth understanding of sex determination and gonadal development in chickens.

It is known that nutrition and immunity are connected, but the mechanism is not very clear. Endogenous retroviruses (ERV) account for 8 to 10% of the human and mouse genomes and play an important role in some biological processes of animals. Recent studies indicate that the activation of ERV can affect the expression of the immunity- or inflammation-related genes, and the activities of ERV are subjected to regulation of many factors including nutritional factors. Therefore, we hypothesize that nutritional status can affect the expression of the immunity- or inflammation-related genes via ERV. To verify this hypothesis, the nutritional status of animals was altered by fasting or overfeeding, and the expression of intact ERV (ERVK18P, ERVK25P) and immunity- or inflammation-related genes (DDX41, IFIH1, IFNG, IRF7, STAT3) in the liver was determined by quantitative PCR, followed by overexpressing ERVK25P in goose primary hepatocytes and determining the expression of the immunity- or inflammation-related genes. The data showed that compared with the control group (no fasting), the expression of ERV and the immunity- or inflammation-related genes was increased in the liver of the fasted chickens but decreased in the liver of the fasted geese. Moreover, compared with the control group (routinely fed), the expression of ERV and the immunity- or inflammation-related genes was increased in the liver of the overfed geese. In addition, overexpression of ERVK25P in goose primary hepatocytes can induce the expression of the immunity- or inflammation-related genes. In conclusion, these findings suggest that ERV mediate the effects of fasting and overfeeding on the expression of the immunity- or inflammation-related genes, the mediation varied with poultry species, and ERV and the immunity- or inflammation-related genes may be involved in the development of goose fatty liver. This study provides a potential mechanism for the connection between nutrition and immunity.

Nutrition and energy are essential for poultry growth and production performance. Fasting and refeeding have been widely used to study the effects of nutrition, energy, and related mechanisms in chicken. Previous studies have shown that geese have a strong capacity for fat synthesis and storage; thus, changes in the goose liver transcriptome may be different from those in chicken assessed with a model of fasting and refeeding. However, the responses of the goose liver transcriptome to fasting and refeeding have not yet been addressed. In this study, 36 70-day-old Si Ji geese with similar body weight were randomly assigned to three groups: control (ad libitum feeding), fasting (fasted for 24 h), and refeeding (fast for 24 h followed by 2-h feeding) groups. After treatment, eight geese per group were sacrificed for sample collection. Liver samples from four geese in each group were subjected to transcriptome analysis, followed by validation of differentially expressed genes (DEGs) using quantitative polymerase chain reaction with the remaining samples. As a result, 155 DEGs (73 up-regulated) were identified between the control and fasting groups, and 651 DEGs (321 up-regulated) were identified between the fasting and refeeding groups. The enrichment analyses of Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways showed that fasting mainly influenced material metabolism in the liver, especially lipid metabolism; in contrast, refeeding affected not only lipid metabolism but also glucose and amino acid metabolism. In addition, the peroxisome proliferator-activated receptor (PPAR) signaling pathway may play an important role in lipid metabolism. In conclusion, fasting and refeeding have a strong effect on lipid metabolism in the goose liver; specifically, fasting promotes fatty acid oxidation and inhibits fatty acid synthesis, and refeeding has the opposite effect. The model of fasting and refeeding is suitable for goose nutrition studies.

Overfeeding causes severe steatosis but not inflammation in goose liver, suggesting existence of protective components. Previous studies have shown that some intestinal microbes and their metabolites damage intestinal structural integrity and function, thus causing inflammation in the development of human and mouse nonalcoholic fatty liver disease. Therefore, this study hypothesizes that intestinal structural integrity of goose is maintained during overfeeding, which may provide goose fatty liver a protective mechanism against inflammation. To test this hypothesis, 48 seventy-day-old healthy Landes male geese were overfed (as overfeeding group) or normally fed (as control group). Blood and intestine (jejunum, ileum, and cecum) samples were harvested on the 12th and 24th d of overfeeding. Data showed that goose fatty liver was successfully induced by 24 d of overfeeding. Hematoxylin-eosin staining analysis indicated that the arrangement of villi and crypts in the intestine was orderly, and the intestinal structure was intact with no pathological symptoms in the 2 groups. Enzyme-linked immunosorbent assay and quantitative PCR analysis indicated no significant differences in the expression of tight junction and inflammation-related genes as well as plasma lipopolysaccharide concentration between the groups. Ileal hypertrophy and cecal atrophy were observed in the overfed vs. control geese, probably because of change of sphingolipid metabolism. Activation of apoptotic pathway may help cecum avoid necrosis-induced inflammation. In conclusion, healthy and intact intestine provides a layer of protection for goose fatty liver against inflammation. Sphingolipid metabolism may be involved in the adaptation of ileum and cecum to overfeeding. The hypertrophy of ileum makes it an important contributor to the development of goose fatty liver. The atrophy and decline in the function of cecum may be caused by apoptosis induced by overfeeding.

Fatty liver hemorrhagic syndrome (FLHS), usually occurring in hens in the late laying period, can lower egg yield and even cause death. Similar to nonalcoholic fatty liver disease in humans, FLHS may begin with simple hepatic steatosis. It is known that gut microbiota is important to the development of nonalcoholic fatty liver disease, but the relationship between FLHS and gut bacteria remains unclear. In this study, bacteria compositions in the ileum and cecum were determined by 16S rDNA sequencing analysis in the groups of hens with vs. without hepatic steatosis (average liver fat contents were 0.34 mmol/g protein or 2.6% vs. 0.84 mmol/g protein or 6.0%). These hens were in the 2 tails of the liver fat content distribution with each tail accounting for 10% of the test population containing 90 66-wk-old healthy Rhode Island Red laying hens raised under routine feeding regimen. The results showed that liver weight but not the weights of the body, heart and abdominal fat were significantly different between the groups. Moreover, bacterial diversity was not significantly different between the groups, but bacterial diversity in the cecum was higher than that in the ileum. Proteobacteria was the most dominant phylum in the ileum, whereas Proteobacteria, Firmicutes, and Bacteroidetes were the most dominant phyla in the cecum. Some bacteria were common to the ileum and cecum, but some were unique. Furthermore, some bacteria were significantly different in abundance between the groups, that is the group without hepatic steatosis had more bacteria inhibiting host energy absorption or benefiting intestinal health, whereas the group with hepatic steatosis had more conditionally pathogenic or harmful bacteria. In conclusion, hepatic steatosis in laying hens is associated with intestinal bacterial composition (bacterial abundance) but not bacterial diversity. The identified common and unique bacteria are related to the functional characteristics of the ileum and cecum. The decrease of beneficial bacteria and the increase of harmful bacteria in the intestine of laying hens may increase FLHS incidence by promoting hepatic steatosis.

Glucose oversupply promotes formation of fatty liver, and fatty liver is usually accompanied with hyperglycemia. However, the mechanism by which glucose promotes formation of fatty liver is not very clear. In this study, fatty liver was successfully induced in Landes goose by 19 days of overfeeding with corn-based feed, the overfed geese had a significantly higher level of blood glucose than the normally fed geese (control group). In goose primary liver cells, high level of glucose promoted fat deposition and induced the expression of SREBF2(or SREBP2), a key regulator of lipid metabolism, and its intronic gene, miR-33. Moreover, overexpression of miRNA-33(miR-33) promotes lipid accumulation in goose primary liver cells. Consistently, miR-33 inhibitor suppressed glucose induced lipid accumulation in liver cells. Interestingly, the relative abundance of miR-33 in goose fatty liver was significantly higher than that in normal liver, while the relative mRNA and protein abundances of CROT, the target gene of miR-33, in goose fatty liver were significantly lower than those in goose normal liver. Taken together, these findings suggest that miR-33 mediates glucose promotion of lipid accumulation in goose primary liver cells, and that glucose participates in formation of goose fatty liver by regulating the expression of miR-33/CROT.

Simple Summary In this study, the effects of dietary supplementation of Clostridium butyricum and/or Bacillus subtilis were determined on growth performance, intestinal antioxidative capacity, intestinal morphology, cytokine production, and intestinal microbial composition in Yangzhou geese. Data showed that probiotics promoted feed intake and growth, improved antioxidative capacity and intestinal morphology, increased the relative abundances of Firmicutes and Lactobacillus in intestinal content, decreased the relative abundances of Proteobacteria and Ralstonia, and altered α-diversity and the predicted functions of intestinal microflora, but did not induce the expression of genes related to intestinal inflammation and tight junction. Abstract Probiotics are a substitute for antibiotics in the sense of intestinal health maintenance. Clostridium butyricum and Bacillus subtilis, as probiotic bacteria, have been widely used in animal production. The aim of this study was to investigate the effects of the two probiotic bacteria in geese. A total of 288 1-day old, healthy Yangzhou geese were randomly assigned into 4 groups (A, B, C and D) with 6 replicates of 12 birds each. Group A, as control, was fed a basal diet, and the treatment groups (B, C and D) were fed the basal diet supplemented with 250 mg/kg Clostridium butyricum (the viable count was 3.0 × 106 CFU/g), 250 mg/kg Bacillus subtilis (the viable count was 2.0 × 107 CFU/g), or a combination of the two probiotic bacteria for 70 days, respectively. The results indicated that: compared with the control group, dietary probiotics (1) promoted the growth and feed intake of the geese, (2) increased the absolute weight of duodenum, (3) increased the antioxidative capacity (total antioxidative capacity (T-AOC), total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-PX)) of intestinal mucosa, (4) improved intestinal morphology (the ratio of villus height to crypt depth), (5) but did not induce inflammation and changes of tight junction in the intestine, which was indicated by no induction of pro/inflammatory cytokines (IL-1β, IL-6, IL-10, TNFAIP3) and tight junction related genes (TJP1 and OCLN). Moreover, dietary probiotics increased the relative abundances of Firmicutes phylum and Lactobacillus genus and decreased the relative abundances of Proteobacteria phylum or Ralstonia genus in the intestinal content. In addition, the alpha diversity (observed species, Chao1, and estimate the number of OTUs in the community(ACE)) was reduced and the predicted functions of intestinal microflora, including peptidases, carbon fixation and metabolic function of starch and sugar, were enhanced by dietary probiotics. In conclusion, dietary probiotics promote the growth of geese by their positive effects on intestinal structure and function, the composition and functions of gut microflora, and intestinal antioxidative capacity.

Selenized glucose can be easily prepared via the selenization reaction of glucose using in situ generated NaHSe as the selenization reagent. The technique has been industrialized to produce the chemical in kilogram scale, making it an easily available material in laboratory presently. The selenized glucose may be widely used as the starting material for the preparation of selenium-containing catalysts, as the organoselenium additive for feeds, and as the efficient selenium-enriched foliar fertilizers. In this work, we found that treating Fusarium graminearum, a fungal pathogen inciting wheat scab disease, with selenium glucose could significantly inhibit the generation of the deoxynivalenol (DON) toxin, which might be a breakthrough for reducing the detriment of the wheat scab disease.

Production of pimpled or sandpaper-shelled eggs (SE) is a major problem in aged hens. Probiotics can improve eggshell quality; however, the relationship between SE production and gut bacteria remains unclear. Here, 1200 450-d-old Hy-line hens were assigned to four groups (300 hens each), with the control group fed basal diet and treatment groups fed basal diet plus 500, 1000, and 1500 mg/kg of

OBJECTIVE This study was conducted to investigate the effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the energy metabolism in thigh muscle of embryos and neonatal broilers. METHODS A total of 960 eggs were randomly assigned to three treatments: i) non-injected control group, ii) saline group injected with 0.6 mL of physiological saline (0.75%), and iii) CrPyr group injected with 0.6 mL of physiologi-cal saline (0.75%) containing 12 mg CrPyr/egg on 17.5 d of incubation. After hatching, 120 male chicks (close to the average body weight of the pooled group) in each group were randomly assigned to eight replications. The feeding experiment lasted 7 days. RESULTS The results showed that IOF of CrPyr increased glucose concentrations in the thigh muscle of broilers on 2 d after injection (p

Background Previous studies indicate that microRNA33 (miR-33) and its target gene, CROT, are implicated in hepatic lipid metabolism, but it is unclear whether miR-33 participates in the development of goose fatty liver via CROT. Methods The expression of miR-33 in goose fatty liver, muscle and fat tissues, as well as the mRNA and protein expression of CROT in goose fatty liver was determined by q-PCR or Western-blot. The targeting regulatory relationship between miR-33 and CROT in goose liver cells was validated by miR-33 overexpression and interference assays. The effects of miR-33 mimic and CROT overexpression on lipid deposition and the expression of downstream genes were determined in goose primary hepatocytes. The treatment of high concentrations of glucose and insulin was performed to determine their regulation on the expression of miR-33 and CROT in goose primary hepatocytes. Results Here, data showed that miR-33 expression was significantly increased in the liver, muscle and fat tissues of overfed geese. Consistently, miR-33 mimic promoted lipid deposition in goose primary hepatocytes. Moreover, the regulatory targeting relationship between miR-33 and CROT was validated in goose primary hepatocytes. Consistently, the mRNA and protein expression of CROT were significantly reduced in goose fatty liver. Interestingly, CROT overexpression could induce the expression of fatty acid oxidation associated genes including CRAT, PEX5, EHHADH, CAT and ACOT8 in goose primary hepatocytes, but only the expression of PEX5 was significantly inhibited in goose fatty liver. However, it seemed conflicting that CROT overexpression increased lipid deposition and reduced lipid peroxidation in goose primary hepatocytes. Additionally, high glucose inhibited miR-33 expression and induced CROT expression in goose primary hepatocytes. Conclusions These findings suggest that miR-33 potentially participates in the development of goose fatty liver via CROT, and that miR-33/CROT may partially mediate the effect of glucose in goose liver cells.

Non-alcoholic fatty liver disease (NAFLD) occurs in humans, domestic animals and poultry. Different from upregulation of complement C3 in human NAFLD, C3 expression is inhibited in goose fatty liver (GFL), implying a specific role of C3 in GFL. This study was mainly focused on uncovering the uniqueness of goose liver cells in the regulation of C3 expression and identifying the downstream genes of C3 to improve understanding on the specific role of C3 in GFL. The results showed that C3 expression was inhibited in the liver, muscle and fat tissues of the overfed versus control (normally fed) geese. Oleate and insulin could inhibit C3 expression in goose primary hepatocytes but induce it in mouse primary hepatocytes. A total of 1,123 differentially expressed genes (DEGs) were affected by C3 overexpression and were mainly enriched in immune response/inflammation and catabolism-related KEGG pathways. Additionally, the representative downstream genes (FASN and ETNK1) of C3 could mediate the role of C3 in the development of GFL. In conclusion, the suppression of C3 in GFL is at least partially attributed to hyperinsulinemia, hyperlipidemia and uniqueness of goose liver cells. Complement C3 does not only affect hepatic steatosis but also affect inflammation/immune response in GFL.

Selenized glucose (SeGlu) is a new type of organic selenium (Se) that is synthesized through the selenide reaction of glucose with sodium hydrogen selenide. This study aimed to clarify the influence of dietary SeGlu on the Se level and antioxidant capacity of the liver, oviduct, and spleen in laying hens. A total of 360, 60-week-old, Hy-Line Brown laying hens were randomly assigned to three treatment groups: a basal diet alone (control group, without adding exogenous Se) or the basal diet supplemented with 0.3 mg/kg of Se from sodium selenite (SS) or 5 mg/kg of Se from SeGlu. Diets with SeGlu increased Se levels in the liver, oviduct, and spleen of laying hens (P < 0.001). Compared with the control and SS groups, diet supplemented with SeGlu enhanced glutathione peroxidase (GSH-Px) activity and total antioxidant capacity (T-AOC) in the spleen and oviduct as well as the scavenging ability of 2, 2-diphenyl-1-picrylhydrazyl free radical (DPPH•) in the oviduct (P < 0.05). Compared with the control group, SeGlu treatment resulted in an increase (P < 0.05) in GSH-Px activity, T-AOC, and scavenging abilities of hydroxyl radical and DPPH• in the liver of hens. In addition, dietary SeGlu and SS decreased the hydrogen peroxide level in the oviduct in comparison to the control group (P < 0.05). Therefore, dietary SeGlu increased Se concentration and antioxidant ability in the liver, oviduct, and spleen of laying hens. Moreover, SeGlu may be used as a potential source of Se additive in laying hen production.

No overt pathological symptoms are observed in the goose liver with severe steatosis, suggesting that geese may host unique protective mechanisms. Gas chromatography time-of-flight mass spectrometry-based metabolomics analyses of liver and intestinal contents in overfed vs. normally fed geese (26 geese in each treatment) were investigated. We found that overfeeding significantly changed the metabolic profiles of liver and intestinal contents. The differential metabolites mainly belong to fatty acids, amino acids, organic acids, and amines. The differential metabolites were involved in glycolysis/gluconeogenesis, glycerolipid metabolism, the pentose phosphate pathway, fatty acid degradation, the sphingolipid signaling pathway, and the biosynthesis of unsaturated fatty acids. Moreover, we determined the biological effects of arachidonic acid (ARA) and tetrahydrocorticosterone (TD) in goose primary hepatocytes and intestinal cells. Data showed that the mRNA expression of arachidonate 5-lipoxygenase (ALOX5) in goose primary intestinal cells was significantly induced by 0.50 mM ARA treatment. Cytochrome P-450 27A1 (CYP27A1) mRNA expression was significantly inhibited in goose primary hepatocytes by 1 &micro, M TD treatment. In conclusion, the formation of goose fatty liver is accompanied by significant changes in the metabolic profiles of liver and intestinal contents, and the changes are closely related to the metabolisms of glucose and fatty acids, oxidative stress, and inflammatory reactions.

Communication between tissues and organs plays an important role in the maintenance of normal physiological functions as well as the occurrence and development of diseases. Communication molecules act as a bridge for interactions between tissues and organs, playing not only a local role in the tissues and organs where they are secreted but also in exerting systemic effects on the whole body via circulation. In this study, blood microRNA-omics analysis of overfed vs. normally fed (control) Landes geese revealed that the content of each of the 21 microRNAs (miRNAs) in the blood of overfed geese was significantly higher than that in the blood of control geese. These miRNAs may have systematic effects in the development of goose fatty liver as well as being candidate markers for the diagnosis of goose fatty liver. We determined the expression of miR-143, miR-455-5p, miR-222a-5p, miR-184, miR-1662, and miR-129-5p using quantitative PCR in goose fatty liver vs. that in normal liver. The expression of these miRNAs, except miR-129-5p, in goose fatty liver was also significantly higher than that in normal liver (P

Goose fatty liver may have a unique protective mechanism as it does not show a pathological injury even in the case of severe steatosis. Although neural precursor cell-expressed developmentally downregulated gene 4 (NEDD4) participates in repair and regeneration of injured liver through its target proteins, its role in nonalcoholic fatty liver disease remains unknown. Using quantitative polymerase chain reaction (PCR) and immunoblot analyses, here, we found that the messenger RNA (mRNA) and protein expressions of NEDD4 were induced in goose fatty liver compared with normal liver. The mRNA expression of the gene of phosphate and tension homology deleted on chromosome ten (PTEN) and insulin-like growth factor 1 receptor (IGF1R) was also induced in goose fatty liver; however, their protein expression was or tended to be suppressed. Moreover, the co-immunoprecipitation analysis indicated that there was a physical association between NEDD4 and PTEN in goose liver, which was consistent with the ubiquitination of PTEN in goose fatty liver. Furthermore, NEDD4 overexpression in goose primary hepatocytes suppressed the PTEN and IGF1R protein levels without a significant effect on their mRNA expression. In conclusion, the increased expression of NEDD4 leads to the degradation of PTEN and IGF1R proteins through ubiquitination in goose fatty liver, suggesting that NEDD4 may protect goose fatty liver from severe steatosis-associated injury via its target proteins during the development of goose fatty liver.

This study aims to investigate in ovo feeding (IOF) of creatine pyruvate (CrPyr) on glucose metabolism, hormone concentration, and the 5'-AMP-activated protein kinase (AMPK) pathway in breast muscle of embryos and neonatal broilers. The three treatments were noninjected control, 0.75% NaCl treatment, and 12 mg CrPyr/egg treatment. The solution was injected on the 17.5 day of incubation. At hatch, 120 male broilers from each treatment were chosen for a 7 day feeding trial. Compared with other treatments, CrPyr treated broilers enhanced insulin and thyroxine levels in plasma, adenosine triphosphate (ATP) concentration, hexokinase and pyruvate kinase activities, glucose transporter protein mRNA expressions, as well as protein abundances of phosphor-liver kinase B1 and phosphor-AMPK in breast muscle at hatch. In conclusion, IOF of CrPyr improved the energy status, increased the gene expression of glucose transporter proteins, and facilitated glycolysis in breast muscle, which may be associated with the activated AMPK pathway.

Probiotics are a substitute for antibiotics in the sense of intestinal health maintenance. Clostridium butyricum and Bacillus subtilis, as probiotic bacteria, have been widely used in animal production. The aim of this study was to investigate the effects of the two probiotic bacteria in geese. A total of 288 1-day old, healthy Yangzhou geese were randomly assigned into 4 groups (A, B, C and D) with 6 replicates of 12 birds each. Group A, as control, was fed a basal diet, and the treatment groups (B, C and D) were fed the basal diet supplemented with 250 mg/kg Clostridium butyricum (the viable count was 3.0 × 106 CFU/g), 250 mg/kg Bacillus subtilis (the viable count was 2.0 × 107 CFU/g), or a combination of the two probiotic bacteria for 70 days, respectively. The results indicated that: compared with the control group, dietary probiotics (1) promoted the growth and feed intake of the geese, (2) increased the absolute weight of duodenum, (3) increased the antioxidative capacity (total antioxidative capacity (T-AOC), total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-PX)) of intestinal mucosa, (4) improved intestinal morphology (the ratio of villus height to crypt depth), (5) but did not induce inflammation and changes of tight junction in the intestine, which was indicated by no induction of pro/inflammatory cytokines (IL-1β, IL-6,&nbsp, IL-10, TNFAIP3) and tight junction related genes (TJP1 and OCLN). Moreover, dietary probiotics increased the relative abundances of Firmicutes phylum and Lactobacillus genus and decreased the relative abundances of Proteobacteria phylum or Ralstonia genus in the intestinal content. In addition, the alpha diversity (observed species, Chao1, and estimate the number of OTUs in the community(ACE)) was reduced and the predicted functions of intestinal microflora, including peptidases, carbon fixation and metabolic function of starch and sugar, were enhanced by dietary probiotics. In conclusion, dietary probiotics promote the growth of geese by their positive effects on intestinal structure and function, the composition and functions of gut microflora, and intestinal antioxidative capacity.

The avian eggshell is formed in the uterus, and eggshell quality usually decreases markedly in the late phase of hen laying cycles. Production of sandpaper-shelled eggs (SE), a category of eggs with relatively less eggshell quality, causes a great economic loss. Underlying mechanisms of SE formation, however, remain unclear. For the present study, it was hypothesized that alterations in uterine structure and function contribute to SE formation. To test this hypothesis, uterine samples were collected from 450-day-old hens that produced normal eggs (NE) and SE (based on 2-week-long assessments, n = 10) for histomorphological and transcriptome analyses. Compared with the NE group, uteri of the SE group were apparently atrophied. Furthermore, a total of 211 differentially expressed genes (DEGs) were identified in the uteri of hens of the two groups. These DEGs were clustered into 145 gene ontology terms (FDR < 0.05) and enriched in 12 KEGG pathways (P < 0.10), which are primarily related to organ morphogenesis and development, cell growth, differentiation and death, ion transport, endocrine and cell communication, immune response, and corticotropin-releasing hormones. In particular, corticotropin may be an important factor in SE formation because of effects on ion transport. Furthermore, as indicated by lesser abundances of relevant mRNA transcripts, the lesser expression of genes related to ion transport and matrix proteins also contribute to SE production because of effects on eggshell formation. In conclusion, results from this study revealed there were structural and functional differences in the hen uterus in NE and SE groups.

During the last phase of incubation, dramatic physiological and metabolic changes occur in chick embryos, and supplies of nutrients and energy are always insufficient. This study investigated the effects of in ovo feeding (IOF) of l-arginine (Arg) on the hatchability, growth performance, intestinal development and functions of posthatch broilers.; Results: The IOF of Arg increased (P

In this study, we evaluated the meat flavor compounds of Yangzhou geese, including one group of pedigree strain (AA group) and 4 groups of 2-strain crossbreds (KA, KB, CA, and SA). Each group consisted of 100 geese comprised of 5 replicates of 10 males and 10 females each. Inosine 5'-monophosphate (IMP), amino acid (AA), and fatty acid (FA) levels in breast and thigh muscle were determined. Results showed that AA group had the highest levels of total amino acid (TAA) and dissolved free amino acids (DFAA) in breast muscle and of polyunsaturated fatty acids (PUFA) in thigh muscle (P0.05). In SA group, the levels of C17:1, C22:0, C22:1, C20:4, and C24:1 in breast muscle were significantly higher in SA than in other groups (P0.05). KB group had the lowest glycine levels in breast muscle (P0.05) while MUFA levels were significantly higher in KB than in other groups (P0.05). In KA, the levels of C18:3 in breast muscle in were higher than in CA and KB (P0.05). CA had relevant higher IMPc levels in breast muscle than SA (P0.05) and other groups (P0.05); however, no significant differences were obtained in thigh muscle (P0.05). In conclusion, Yangzhou AA goose has high levels of meat flavor compounds than its crossbreeds. Future efforts should focus on assessing meat flavor through measurement of sensory characteristics of Yangzhou geese.

During the last phase of incubation, dramatic physiological and metabolic changes occur in chick embryos, and supplies of nutrients and energy are always insufficient. This study investigated the effects of in ovo feeding (IOF) of l-arginine (Arg) on the hatchability, growth performance, intestinal development and functions of posthatch broilers.The IOF of Arg increased (P 0.05) the feed intake and body weight gain during 1-21 days and 1-42 days, and the intestinal weight of 7- and 21-day-old broilers, compared with non-injected control and diluent-injected groups. The IOF of Arg increased (P 0.05) villus height (VH), ratio of VH to crypt depth (CD) and density of goblet cells, and decreased (P 0.05) the CD in jejunum of 1-, 7- and 21-day-old broilers. The IOF of Arg also increased (P 0.05) the percentage of proliferating cell nuclear antigen positive cells of villus, and the mRNA expressions of mucin-2, claudin-1, zonula occludens-1 and -2 in jejunal mucosa of 21-day-old broilers. Meanwhile, IOF of Arg increased (P 0.05) the protein abundance of phosphorylated mechanistic target of rapamycin (mTOR), ribosomal protein S6 kinase 1 and eukaryotic initiation factor 4E binding protein 1 in jejunal mucosa.The IOF of Arg improved the development and barrier functions of small intestine, which might be associated with activating the mTOR pathway. In addition, the improved intestinal development might explain the improvement in feed intake and consequently the growth performance of broilers. Therefore, IOF of Arg solution could be an effective technology for regulating early nutrition supply and subsequent growth development in the poultry industry. © 2017 Society of Chemical Industry.

Summary The effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the growth performance, energy reserves and mRNA expression levels of gluconeogenesis and glycogenesis enzymes in liver of late-term embryos and neonatal broilers were investigated. After candling on 16 day of incubation, a total of 960 eggs were randomly assigned to three treatments: (i) non-injected control, (ii) saline group injected with 0.6 ml of 0.75% physiological saline and (iii) Creatine pyruvate group injected with 0.6 ml of physiological saline containing 12 mg CrPyr/egg. After hatching, 120 male chicks with average body weight (BW) were randomly allocated into each treatment group for a 7-day feeding trial. The results showed that broilers subjected to CrPyr treatment had higher BW than those of the control and saline groups on 1, 3 and 7 day post-hatch, as well as the yolk sac weight on 19 day of incubation (19 E), the day of hatch and 3 day post-hatch (p

Summary This study was to investigate the effects of in ovo feeding (IOF) L-arginine (Arg) solution on the development of digestive organs, the duodenal mucosa of broiler embryos and hatchlings, and the growth performance of chicks during the first week post-hatch. A total of 720 fertilized eggs with similar weight were randomly allocated to three groups, consisting of eight replicates of 30 eggs each. Three treatments were arranged as non-injected control, diluent-injected (0.75% NaCl solution) group and Arg solution-injected group containing 1% Arg, dissolved in diluent. At 17.5 days of incubation, 0.6 ml of IOF solution was injected into amniotic fluid of each egg of injected groups. Results showed IOF of Arg solution increased (p