Parental betaine supplementation promotes gosling growth with epigenetic modulation of IGF gene family in the liver.

Betaine is widely used as a feed additive in the chicken industry to promote laying performance and growth performance, yet it is unknown whether betaine can be used in geese to improve the laying performance of goose breeders and the growth traits of offspring goslings. In this study, laying goose breeders at 39 wk of age were fed basal (Control, CON) or betaine-supplemented diets at low (2.5 g/kg, LBT) or high (5 g/kg, HBT) levels for 7 wk, and the breeder eggs laid in the last week were collected for incubation. Offspring goslings were examined at 35 and 63 d of age. The laying rate tended to be increased (P = 0.065), and the feed efficiency of the breeders was improved by betaine supplementation, while the average daily gain of the offspring goslings was significantly increased (P < 0.05). Concentrations of insulin-like growth factor 2 (IGF-2) in serum and liver were significantly increased in the HBT group (P < 0.05), with age-dependent alterations of serum T3 levels. Concurrently, hepatic mRNA expression of the IGF gene family was significantly increased in goslings derived from betaine-treated breeders (P < 0.05). A higher ratio of proliferating cell nuclear antigen (PCNA)-immunopositive nuclei was found in the liver sections of the HBT group, which was confirmed by significantly upregulated hepatic expression of PCNA mRNA and protein (P < 0.05). Moreover, hepatic expression of thyroxine deiodinase type 1 (Dio1) and thyroid hormone receptor ß (TRß) was also significantly upregulated in goslings of the HBT group (P < 0.05). These changes were associated with significantly higher levels of global DNA 5-mC methylation, together with increased expression of methyl transfer genes (P < 0.05), including betaine-homocysteine methyltransferase (BHMT), glycine N-methyltransferase (GNMT), and DNA (cytosine-5-)-methyltransferase 1 (DNMT1). The promoter regions of IGF-2 genes, as well as the predicted TRß binding site on the IGF-2 gene, were significantly hypomethylated (P < 0.05). These results indicate that gosling growth can be improved by dietary betaine supplementation in goose breeders via epigenetic modulation of the IGF gene family, especially IGF-2, in the liver.

The goose industry plays important roles in economics, cultures, and ecosystems, yet the low laying and growth rates of many indigenous breeds hinders the development of the goose farming. Betaine, an important methyl donor, is commonly used as a feed additive in livestock and poultry to enhance animal growth. Dietary supplementation of betaine in laying hens or gestational sows has been reported to promote the growth of their offspring. Here, we sought to investigate whether and how dietary betaine supplementation affects the growth and development of offspring goslings. In this study, goose breeders, both male and female, were fed a basal diet supplemented respectively with 0, 2.5, or 5 g/kg betaine for 7 wk. Goslings hatched from the breeder eggs of different groups were raised under the same standard condition for assessing the growth performance. Parental betaine increases the growth rate of offspring goslings with decreased DNA methylation on the IGF-2 gene promoter and increased expression of the IGF-2 gene in the liver. These results provide scientific evidence for the inter-generational effect of betaine on gosling growth.

Effects of dietary vitamin K3 supplementation on production performance, egg quality, vitamin K-dependent proteins, and antioxidant properties in breeding geese during the laying period.

The aim of this work was to investigate the effects of dietary vitamin K3 (VK3) supplementation on production performance, egg quality, vitamin K-dependent proteins, and antioxidant properties in breeding geese during the laying period. A total of one hundred twenty 82-wk-old Wulong geese with similar body weights were randomly divided into 6 groups with 4 replicates and 5 geese each (1 male and 4 female). The geese in the control group were fed a basal diet, and the geese in the treatment groups were fed diets supplemented with different levels of VK3 (2.5, 5.0, 7.5, 10.0, and 12.5 mg/kg) for 11 wk. Dietary VK3 supplementation linearly and quadratically increased feed intake, egg mass, egg weight, and egg production (P < 0.05). Increasing VK3 levels linearly and quadratically increased albumen height, shell thickness and Haugh unit of eggs (P < 0.05). VK3 reduced osteocalcin (OC) and uncarboxylated osteocalcin (ucOC) levels in the serum. Dietary VK3 addition linearly decreased serum malondialdehyde (MDA) levels (P < 0.01). There was linear and quadratic effect in the activity of serum total superoxide dismutase (T-SOD) (P < 0.01), and linear effect in serum total antioxidant capacity (T-AOC) (P < 0.01). In conclusion, dietary VK3 supplementation enhanced the production performance, egg quality, vitamin K-dependent proteins, and antioxidant properties in breeding geese during the laying period. The optimal dose of dietary VK3 supplementation was 10.0 mg/kg.

Effects of dietary arginine supplementation on production performance, serum biochemicals, antioxidant capacity, and immunity of laying Wulong geese.

This study investigated the effects of dietary arginine supplementation on the production performance, serum biochemicals, antioxidant capacity, and immunity of laying Wulong geese. A total of 150 Wulong geese (34-wk old) with similar body weights were randomly divided into 6 groups with 5 replicates and 5 geese each (1 male and 4 female). The geese in the control group were fed a corn-rapeseed meal basal diet, and the geese in the treatment groups were fed the basal diet supplemented with 0.1, 0.2, 0.3, 0.4, and 0.5% arginine. The experiment lasted for 17 wk. Our results showed that dietary arginine increased the egg production rate (LR) and average egg weight (AEW) of geese in a quadratic manner (P < 0.05). Dietary arginine had a quadratic effect on the contents of total protein (TP) and triglyceride (TG) (P < 0.05) in the serum. Dietary arginine quadratically decreased the content of malondialdehyde (MDA) and increased the activity of total superoxide dismutase (T-SOD) (P < 0.05). Dietary arginine supplementation linearly and quadratically increased the contents of immunoglobulin A (IgA) and immunoglobulin G (IgG), and linearly increased the content of nitric oxide (NO) (P < 0.05). In conclusion, dietary arginine supplementation can significantly improve the production performance, serum biochemicals, antioxidant capacity, and immunity of laying Wulong geese. Therefore, 0.3% arginine (actual content: 1.02%) is recommended in the diet.

Effects of cottonseed meal on performance, gossypol residue, liver function, lipid metabolism, and cecal microbiota in geese.

A total of 240 28-d-old male goslings were used to investigate the effects of cottonseed meal (CSM) on performance, gossypol residue, liver function, lipid metabolism, and cecal microbiota. All birds were randomly allotted into five groups (eight goslings/replicate, six replicates/group) and subjected to a 35-d experiment. Five isonitrogenous and isoenergetic diets were formulated to produce diets in which 0% (control), 25% (CSM25), 50% (CSM50), 75% (CSM75), and 100% (CSM100) of protein from soybean meal was replaced by protein from CSM. The free gossypol contents in the five diets were 0, 44, 92, 135, and 183 mg/kg, respectively. Dietary CSM did not affect the growth performance from 29 to 63 d and carcass traits at 63 d (P > 0.05). Liver gossypol residues were influenced (P < 0.05) by dietary CSM and increased linearly (P < 0.05) and quadratically (P < 0.05) as dietary CSM increased. The malondialdehyde content of the liver was lower in the CSM100 group than in the other groups (P < 0.05). Serum triglyceride and low-density lipoprotein cholesterol were influenced (P < 0.05) by dietary CSM and increased linearly (P < 0.05) with increasing dietary CSM. Dietary CSM altered (P < 0.05) the composition of some fatty acids in the liver and breast muscle. The concentration of linolenic acid and Σn-3 polyunsaturated fatty acid (PUFA) in the liver and breast muscle decreased linearly, but the Σn-6/Σn-3 PUFA ratio increased linearly with increasing dietary CSM (P < 0.05). Dietary CSM affected (P < 0.05) the hepatic gene expression of fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), and apolipoprotein B (ApoB). As the dietary CSM concentration increased, the hepatic gene expression of FAS increased linearly (P < 0.05) and quadratically (P < 0.05), but the hepatic gene expression of ACC and ApoB increased linearly (P < 0.05). The CSM diet decreased the relative abundance of the Bacteroidota and Bacteroides (P < 0.05), and the CSM50 diet increased the relative abundance of the Firmicutes and Colidextribacter (P < 0.05) compared to the control group. Overall, these results show that dietary CSM has no adverse effects on the performance of goslings from 29 to 63 d. However, CSM affected organismal lipid metabolism, reduced products' edible value, and adaptively altered cecum microbiota.

The shortage of feed resources and the rising price have become one of the significant challenges for animal husbandry worldwide. Considering the strong tolerance and adaptability to roughage of geese, less expensive crop byproducts are used in goose feed by animal nutritionists. Cottonseed meal (CSM) is a potential substitute for soybean meal, and the main concern for its use in poultry feed is free gossypol. This study aimed to investigate the effects of CSM on the performance, gossypol residue, liver function, lipid metabolism, and cecal microbiota in geese. Results showed that dietary CSM has no adverse effects on the performance and liver function of goslings. However, gossypol residue in goose liver increased with increasing dietary CSM. Besides, CSM affected organismal lipid metabolism, altered the tissue fatty acid composition, and adaptively changed cecum microbial microbiota. In summary, CSM is a good dietary protein source for geese, but further attention may be needed to its use for the edible value of goose products.

Different amino acid supplementation patterns in low-protein diets on growth performance and nitrogen metabolism of goslings from 1 to 28 days of age.

The investigation aimed to explore the suitable amino acid (AA) supplementation pattern for goslings under low-protein diets. A total of 364 1-day-old male goslings were randomly divided into 4 experimental groups, with 7 pens containing 13 goslings each. The 4 groups were control (CP, 18.55%), LPM (CP, 15.55% + major AA), LPA (CP, 15.55% + all AA), and LPR (CP, 15.55% + AA content reduced proportionally to the control's CP). The corn-soybean meal diets are formulated according to the ideal AA model of goose and its nutritional requirements. The results indicated that the ADG and BW were the lowest, and the F: G was the highest in LPR (P < 0.05); the other three groups were not significantly different (P > 0.05). The ADFI and mortality were not different among all the groups (P > 0.05). Among the AA content in serum and breast muscle, lysine in serum significantly decreased compared with the control (P < 0.05). The UREA content was approximately 2-fold higher in the LPR group than in the LPM and LPA groups (P < 0.05). No difference in IgA, IgG, IgM, and IgE levels was observed among the groups (P > 0.05). The nitrogen excretion was decreased in LPM and LPA compared to the control and LPR (P < 0.05). Nitrogen deposition did not differ among groups (P > 0.05). Nitrogen utilization was highest in the LPA and LPM groups, followed by the control group and LPR (P < 0.05). In conclusion, the patterns of supplementation of major AA and all AA in low-protein diets (CP, 15.55%) had no adverse effect on the growth performance compared with the control (CP, 18.55%) of the goslings. Besides, the two patterns could decrease nitrogen excretion and increase nitrogen utilization. Furthermore, from the perspective of dietary cost and environmental protection, the pattern of supplementing major AA in a corn-soybean meal low-protein diet is suggested.

Isolation and Identification of Antioxidative Peptide from Goose Liver Hydrolysate to Ameliorate Alcohol-Mediated Oxidative Stress Damage in HHL-5 Hepatocytes.

The aim of this study was to isolate and identify antioxidative peptide from goose liver hydrolysate (GLHP) for ameliorating oxidative stress damage by alcohol in HHL-5 hepatocytes. In this research, the target antioxidative peptides in GLHP were separated, purified, and identified via a tangential flow ultrafiltration system combined with size exclusion chromatography (SEC), ion exchange chromatography (IEC), reversed-phase liquid chromatography (RP-LC), and LC-MS/MS. The results suggested that the amino acid sequence of the target antioxidative peptide for ameliorating alcohol-mediated oxidative stress damage in HHL-5 hepatocytes was Leu-Pro-Leu-Pro-Phe-Pro (LPLPFP), which had a molecular weight of 683.41 Da, and was derived from NADH-ubiquinone oxidoreductase chain 1 in goose liver. In addition, LPLPFP was confirmed to have a satisfactory stability and maintained high hepatic protective activity in a simulated gastrointestinal digestion. Moreover, the mechanism of LPLPFP prevented against oxidative stress damage in HHL-5 hepatocytes was attributed to inhibiting the production of reactive oxide species (ROS) by upregulating genes expression in the Ahr-NQO1 signal pathway. In conclusion, these results indicated that dietary GLHP supplementation could ameliorate alcohol-mediated oxidative stress damage and provide an affordable dietary intervention strategy to prevent alcohol-mediated hepatocyte damage.

Effects of cottonseed meal on growth performance, liver redox status, and serum biochemical parameters in goslings at 1 to 28 days of age.

BACKGROUND: Cottonseed meal (CSM), a relatively rich source of protein and amino acids, is used as an inexpensive alternative to soybean meal (SBM) in poultry diets. However, the toxicity of free gossypol in CSM has been a primary concern. The present study was conducted to investigate the effects of CSM on growth performance, serum biochemical parameters, and liver redox status in goslings at 1 to 28 days of age. Three hundred 1-day-old male goslings were randomly divided into 5 groups (10 goslings/pen, 6 replicate pens/group) and subjected to a 28-day experiment. Five isonitrogenous and isoenergetic diets were formulated such that 0% (control), 25% (CSM25), 50% (CSM50), 75% (CSM75), and 100% (CSM100) of protein from SBM was replaced by protein from CSM. The free gossypol contents in the five diets were 0, 56, 109, 166, and 222 mg/kg, respectively. RESULTS: The results showed that dietary CSM was associated with linear decreases in body weight, average daily feed intake and average daily gain and linear increases in the feed-to-gain ratio from 1 to 28 days of age (P < 0.001). As the dietary CSM concentration increased, a numerical increase was found in the mortality of goslings. According to a single-slope broken-line model, the breakpoints for the average daily gain of dietary free gossypol concentration on days 1 to 14, 15 to 28, and 1 to 28 occurred at 23.63, 14.78, and 18.53 mg/kg, respectively. As the dietary CSM concentration increased, serum albumin (P < 0.001) concentrations decreased linearly and serum uric acid (P = 0.011) increased linearly. The hydroxyl radical scavenging ability (P = 0.002) and catalase (P < 0.001) and glutathione peroxidase (P = 0.001) activities of the liver decreased linearly with increasing dietary CSM. However, dietary CSM did not affect the concentrations of reactive oxygen metabolites, malondialdehyde, or protein carbonyl in the liver. CONCLUSIONS: The increasing dietary CSM increased the concentration of free gossypol and altered the composition of some amino acids in the diet. A high concentration of CSM reduced the growth performance of goslings aged 1 to 28 days by decreasing feed intake, liver metabolism, and antioxidant capacity. From the primary concern of free gossypol in CSM, the tolerance of goslings to free gossypol from CSM is low, and the toxicity of free gossypol has a cumulative effect over time.

Effects of betaine on growth performance, intestinal health, and immune response of goslings challenged with lipopolysaccharide.

The objective of this experiment was to investigate the effects of betaine on growth performance, serum parameters, intestinal health, and immune performance of goslings in response to lipopolysaccharide (LPS) challenge. A total of 168 healthy male 15-day-old Jiangnan White Goslings were randomly divided into 4 groups, with 6 replicates per treatment and seven goslings per replicate. A 2 × 2 factorial arrangement included 2 factors, that is, LPS challenge (injection of LPS or physiological saline) and betaine (added 0 or 0.06% betaine in diet). The results indicated that LPS challenge significantly reduced the average daily feed intake (ADFI), average daily gain (ADG), and body weight (BW) at 21 D of the goslings, while dietary betaine supplementation tended to increase the ADFI during the LPS stress period (P = 0.08) and BW at 21 D of the goslings (P = 0.09). The LPS-challenged goslings showed higher pro-inflammatory cytokines (interleukin-1 [IL-1ß], interleukin-6 [IL-6], tumor necrosis factor-α (TNF-α), and Interferon-gamma [IFN-γ]) and lower anti-inflammatory cytokine (Interleukin-10 [IL-10]) (P < 0.05) at 21 D of age. Dietary betaine supplementation alleviated LPS-induced increase in pro-inflammatory cytokines. The LPS challenge significantly decreased duodenal and jejunal villus height (VH) and villus height and crypt depth ratio (VCR), while the addition of betaine significantly increased duodenal VH and VCR (P < 0.05). On the other hand, addition of betaine significantly alleviated decline of enzyme activity on lipase, amylase, trypsin, and chymotrypsin in the intestinal of goslings. The LPS challenge significantly increased the content of serum D-lactic acid (D-LA) and the activity of diamine oxidase (DAO) at 21 D of the goslings. The LPS challenge and betaine addition significantly increased the mRNA expression of Occcludin (OCLN) in jejunal mucosa at 28 D of the goslings (P < 0.05). In conclusion, our research demonstrated that betaine can alleviate the decline of growth performance and immune performance in goslings caused by LPS. The results also indicate betaine possesses anti-inflammation properties and improves intestinal barrier functions. We recommend that 0.06% betaine be added into the diet to improve the intestinal health and immune performance of goslings.

Effects of zinc and Bacillus subtilis on the reproductive performance, egg quality, nutrient digestion, intestinal morphology, and serum antioxidant capacity of geese breeders.

The effects of zinc (Zn) and Bacillus subtilis (B. subtilis) on reproductive performance, egg quality, nutrient digestion, intestine morphology, and antioxidant capacity were explored in geese breeders. Geese breeders (n = 120, 46-wk of age) were randomly assigned into 6 groups with 4 replicates of 5 birds each (1 male and 4 female). Breeders were fed diets with 2 levels of B. subtilis (2.5 × 109 and 5 × 109 CFU/kg) crossed with three levels of Zn (25, 45, and 65 mg/kg) for duration of 10-wk. The results showed that the egg laying rate (P < 0.05), fertility rate (P < 0.01), hatchability rate (P < 0.05), yolk color (P < 0.05), and the retentions of crude protein (P < 0.05), ether extract (P < 0.05) and phosphorus of geese breeders were improved by dietary supplementation of 5 × 109 CFU/kg B. subtilis and 25 mg or 45 mg/kg Zn. The serum T-SOD (P < 0.05) was increased by 45 mg/kg Zn supplementation. The serum T-AOC (P < 0.05) and retention of Zn (P < 0.05) were increased by 5 × 109 CFU/kg B. subtilis supplementation. The birds fed with 5 × 109 CFU/kg B. subtilis and 25 mg or 45 mg/kg Zn showed improved villus length (P < 0.01) and villus length/ crypt depth (P < 0.01) in both the jejunum and ileum. In conclusion, the combination of B. subtilis and Zn may have synergistic effects on these parameters, and dietary inclusion of 5 × 109 CFU/kg B. subtilis and 45 mg/kg Zn is recommended for improving the reproductive performance of geese breeders.

Effects of dietary supplementation of chitosan on immune function in growing Huoyan geese.

This present experiment was performed to investigate the effects of dietary supplementation of chitosan (CS) on immune function in growing Huoyan geese. A total of 320 28-day-old healthy growing Huoyan geese (sex balance) with similar body weight were randomly allotted into control, CS100, CS200, and CS400 groups. Each group includes 4 replicates with 20 geese per replicate, and the feeding trial lasted for 4 wk. The 4 diets contained 0, 100, 200, and 400 mg CS per kg feed, respectively. The results showed that compared with the control group, the relative weight of thymus, serum concentrations of IGF-I, INS, GH, T3, T4, IgM, IgG, IgA, complement C3, and IL-2 in CS200 group were significantly higher at both 42 and 56 D of age, respectively (P < 0.05). In addition, relative weight of bursa of fabricius (BF), spleen, serum complement C4, and TNF-a concentrations in CS200 group were higher at 56 D of age (P < 0.05), no differences were observed at 42 D of age (P > 0.05). These results indicated that addition of 200 mg/kg CS enhanced immune organs weight, serum concentrations of immunoglobulins, complements, hormone, as well as cytokines, and improved immune function of growing Huoyan geese.

The effect of dietary vitamin A supplementation in maternal and its offspring on the early growth performance, liver vitamin A content, and antioxidant index of goslings.

This study investigated the effect of dietary VA supplementation on maternal and its offspring in terms of the early growth performance, antioxidant index, and tissue VA content of the goslings. Yangzhou geese aged 180 D were selected and randomly distributed into 5 experimental groups with 15 female geese and 3 male geese in each group. The geese were fed a basal diet supplemented with 0, 4,000, 8,000, 12,000, or 16,000 IU/kg VA. Eggs were collected from each group starting at 300 D. After hatching, 96 goslings were selected from each maternal group and randomly distributed into 2 experimental groups with factorial arrangement (6 replicates × 8 geese), including 2 levels of VA supplementations, 0 and 9,000 IU/kg. The results are as follows: (1) Different levels of maternal VA supplementation significantly affected the BW and weight gain of 7-day-old offspring (P < 0.05). The weight gain of offspring administered 9,000 IU/kg VA was significantly higher than that of offspring administered the basal diet (P < 0.05). (2) Maternal VA levels significantly affected the T3, T4, and insulin levels of the offspring (P < 0.05). (3) The GSH-PX, SOD, T-AOC, CAT, and tissue VA content of the offspring were significantly higher and MDA was significantly lower in the 9,000 IU/kg VA group than in the no VA group (P < 0.05). (4) Maternal VA levels had a significant effect on offspring GSH, GSH-PX, SOD, MDA, T-AOC, and CAT (P < 0.05). Maternal and offspring VA supplementation interact with the weight gain, tissue VA content, GSH, GSH-PX, SOD, MDA, and CAT of goslings (P < 0.05). Maternal supplementation with 12,000 IU/kg VA and offspring supplementation with 9,000 IU/kg VA was conducive to gosling growth.

Fermented feed regulates growth performance and the cecal microbiota community in geese.

This study was designed to investigate the effects of fermented feed diets on the growth performance and cecal microbial community in geese, and to examine associations between the gut microbiota and growth performance. A total of 720 healthy, 1-day-old male SanHua geese were used for the 55-D experiment. Geese were randomly divided into 4 groups, each with 6 replicates of 30 geese. Groups were fed a basal diet supplemented with 0.0, 2.5, 5.0, or 7.5% fermented feed. The results showed that 7.5% fermented feed had an increasing trend in the body weight and average daily gain of the geese; however, there was no significant response to increasing dietary fermented feed level with regards to ADFI and FCR. In addition, compared with the control group, there was a higher abundance of bacteria in the phylum Bacteroidetes in the cecal samples of geese in the 7.5% fermented feed group (53.18% vs. 41.77%, P < 0.05), whereas the abundance of Firmicutes was lower in the 7.5% fermented feed group (36.30% vs. 44.13%, P > 0.05). At the genus level, the abundance of Bacteroides was increased by adding fermented feed to geese diets, whereas the abundances of Desulfovibrio, Phascolarctobacterium, Lachnospiraceae_uncultured, Ruminiclostridium, and Oscillospira were decreased. These results indicate that fermented feeds have an important effect on the cecal microflora composition of geese, and may affect host growth, nutritional status, and intestinal health.

Dietary selenomethionine increases antioxidant capacity of geese by improving glutathione and thioredoxin systems.

A total of 200 healthy 28-day-old male Jiangnan White geese were used to investigate the free radical scavenging ability, reduced glutathione (GSH) and thioredoxin systems, and the concentrations of reactive oxygen metabolites (ROM), malondialdehyde (MDA), and protein carbonyl (PC) in geese fed diets with organic selenium (Se) (Selenomethionine, SeMet) and inorganic Se (sodium selenite, SS). All geese were randomly allotted into 4 groups with 5 replicates of 10 geese each, and received basal diet supplemented with 0.3 mg Se/kg SS, 0.2, 0.3 and 0.4 mg Se/kg SeMet until 70 D of age, respectively. Geese in the SS and SeMet groups exhibited similar growth performance. Diet with SeMet increased the scavenging abilities of 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt free radical (ABTS•+, P < 0.001) and superoxide radical (O2-•, P = 0.002) in the serum of geese, as well as the scavenging abilities of ABTS•+ (P = 0.023), hydroxyl radical (P = 0.009) and O2-• (P = 0.019) in the liver of geese. Compared to the SS group, SeMet increased hepatic GSH concentration (P = 0.002), the activities of glutathione peroxidase (P = 0.031), γ-glutamate cysteine ligase (P < 0.001), and thioredoxin reductase (P < 0.001), and decreased the concentrations of ROM, MDA, and PC in the serum and liver of geese (P < 0.05). In conclusion, dietary SeMet inclusion would be more effective than SS in increasing the antioxidant capacity of geese, possibly by improving GSH and thioredoxin systems, and 0.2 mg Se/kg SeMet in goose diet is recommended.

The role of active immunization against inhibin α-subunit on testicular development, testosterone concentration and relevant genes expressions in testis, hypothalamus and pituitary glands in Yangzhou goose ganders.

The present study was designed to investigate the potential role of immunization against inhibin on testicular development, plasma testosterone concentration and expression of relevant genes in hypothalamus, pituitary, Leydig and Sertoli cells in Yangzhou ganders. For this purpose, Yangzhou ganders, n = 30 were divided into groups A and B. Group B ganders were actively immunized against inhibin α-subunit, while group A ganders were immunized with bovine serum albumin (BSA), which served as control. Immunization against inhibin elevated testes weights. In addition, immunization against inhibin elevated GnRH, StAR, CYP11A1 and AMH mRNA transcription expressions as depicted by qRT-PCR. Furthermore, hypothalamic GnRH-I mRNA expressions were up regulated, while GnIH mRNA transcription expression showed reciprocal expression on day 227. LH-ß mRNA transcription expression remained unaffected. In conclusion, our findings suggest that active immunization against inhibin affect spermatogenesis and testicular development through regulations of hypothalamic, pituitary and testicular genes expressions.

Transcriptome analysis of hepatic gene expression and DNA methylation in methionine- and betaine-supplemented geese (Anser cygnoides domesticus).

Dietary methionine (Met) restriction produces a coordinated series of transcriptional responses in the liver that limits growth performance and amino acid metabolism. Methyl donor supplementation with betaine (Bet) may protect against this disturbance and affect the molecular basis of gene regulation. However, a lack of genetic information remains an obstacle to understand the mechanisms underlying the relationship between Met and Bet supplementation and its effects on genetic mechanisms. The goal of this study was to identify the effects of dietary supplementation of Met and Bet on growth performance, transcriptomic gene expression, and epigenetic mechanisms in geese on a Met-deficient diet. One hundred and fifty 21-day-old healthy male Yangzhou geese of similar body weight were randomly distributed into 3 groups with 5 replicates per treatment and 10 geese per replicate: Met-deficient diet (Control), Control+1.2 g/kg of Met (Met), and Control+0.6 g/kg of Bet (Bet). All geese had free access to the diet and water throughout rearing. Our results indicated that supplementation of 1.2 g/kg of Met in Met-deficient feed increased growth performance and plasma homocysteine (HCY) levels, indicating increased transsulfuration flux in the liver. Supplementation of 0.6 g/kg Bet had no apparent sparing effect on Met needs for growth performance in growing geese. The expression of many genes critical for Met metabolism is increased in Met supplementation group. In the Bet-supplemented group, genes involved in energy production and conversion were up-regulated. Dietary supplementation with Bet and Met also altered DNA methylation. We observed changes in the methylation of the LOC106032502 promoter and corresponding changes in mRNA expression. In conclusion, Met and Bet supplementation in geese affects the transcriptional regulatory network and alters the hepatic DNA methylation of LOC106032502.

Comprehensive analysis of Sichuan white geese (Anser cygnoides) transcriptome.

High-throughput RNA sequencing was performed for comprehensively analyzing the transcriptome of geese. A total of 28,803,759 bp of raw sequence data was generated by 454 GS Flx+. After removal of adaptor sequences, 28,730,361 bp remained and 117,279 reads were obtained, with an average length of 244 bases. Simultaneously, complementary DNA samples from two different reproductive stages of goose ovarian, hypothalamus and pituitary tissue were sequenced separately using Illumina MiSeq platform. A total of 12 688 673 148 bp of raw sequence data were generated by Illumina MiSeq. After removal of adaptor sequences, 8 198 126 562 bp remained and 60 382 786 clean reads were obtained, with an average length of 135 bases. Assembly of all the reads from both 454 Flx+ and Illumina platforms formed 56,839 contigs. The sequence size ranges from 38 to 28,206 bp in size, with an average size of 2584 bp and an N50 of 4624. The assembly produced a substantial number of large contigs: 35,545 (62.5%) were longer than 1 kb, of which 8850 (15.6%) were longer than 5 kb. The sequencing depth was 85 X on average. We performed comprehensive function annotations on unigenes including protein sequence similarity, gene ontology (GO) term classification, and Kyoto Encylcopedia of Genes and Genomes (KEGG) pathway enrichment. GO analysis showed that approximately 63% of the contigs had annotation information, among the 35,953 annotated isotigs in Nr database, 24,783 (68.9%) sequences were assigned with one or more GO terms. There were 14,634 (40.7%) isotigs for biological processes, 10,557(29.3%) isotigs for cellular component, 22,607 (62.9%) isotigs for molecular function. The result of KEGG pathway mapping 8926 sequences had the pathway annotation, and took part in 477 pathways. Additionally, 10,685 simple sequence repeat (SSR) markers were identified from the assembled sequences. The most frequent repeat motifs were trinucleotides, which accounted for 53.03% of all SSRs, followed by dinucleotides (39.9%), tetranucleotides (5.08%), pentanucleotides (1.68%) and hexanucleotides (0.32%). Transcriptome sequencing on mixture issue of the geese yielded substantial transcriptional sequences and potentially useful SSR markers which provide an important data source for geese research.

Goose fat, a promising nutrient for fish feeding, activates antioxidant enzymes in rainbow trout, Oncorhynchus mykiss.

The objective of this experiment was to test effects of different dietary lipids in rainbow trout feeding on the activity and expression of antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione S-transferase (GST). Four iso-nitrogenous and iso-lipidic casein-gelatin based experimental diets were formulated. The sources of dietary lipids were cod liver oil (CO, rich in polyunsaturated fatty acids), goose fat (GF, rich in saturated fatty acids and monounsaturated fatty acids), soybean oil (SO, rich in linoleic acid), and a blend of CO, GF and SO. Dietary treatments had no significant effect on growth performance and survival was not affected. SOD, GPx and GST enzymes had the maximum activity in GF diet. However qPCR data showed that SOD and GPx mRNA levels were minimum in GF group. Overall data showed that rainbow trout liver enzymes were activated upon GF diet probably activating the enzyme structure itself without stimulating gene expression.

Effects of palmitic acid on lipid metabolism homeostasis and apoptosis in goose primary hepatocytes.

Studies have shown that not only does palmitic acid promote triglyceride (TG) accumulation, but it also affects cell viability in in vitro steatosis models. However, to what degree these effects are mediated by steatosis in goose primary hepatocytes is unknown. In this study, the effects of palmitic acid on the lipid metabolism homeostasis pathway and on apoptosis were determined. The authors measured the mRNA levels of genes involved in TG synthesis, lipid deposition, fatty acid oxidation and the assembly and secretion of VLDL-TG in goose primary hepatocytes. The results indicated that palmitic acid can significantly reduce the activity of goose hepatocytes, and that palmitic acid had a significant effect on TG accumulation; however, with increasing palmitic acid concentrations, the extracellular TG and extracellular VLDL concentration gradually decreased. With increasing palmitic acid concentrations, the gene expression levels of DGAT1, DGAT2, PPARα, CPT-1, FoxO1 and MTTP (which regulate hepatic TG synthesis, fatty acid oxidation and the assembly and secretion of VLDL-TGs) first increased and then decreased; the change in PLIN gene expression was palmitic acid dose-dependent, similar to the regulatory mode of intracellular TG accumulation. In conclusion, this study clearly shows that palmitic acid can promote TG accumulation and induce apoptosis in goose primary hepatocytes, and this effect may be related to the lipid metabolism pathway.

The influence of caecectomy on amino acid availability of three feedstuffs for ganders.

1. Experiments were conducted to assess the influence of caecectomy on amino acid availability (AAA) of three feedstuffs for goose. 2. Nine caecectomised and 9 intact Yangzhou ganders, 24 weeks old, were used in these experiments. Fish meal, soybean meal and cottonseed meal were used as the sole source of protein. The endogenous amino acid (AA) losses were evaluated by a nitrogen (N)-free diet method. The influence of caecectomy on apparent amino acid availability (AAAA) in fish meal, soybean meal and cottonseed meal was assessed in experiment 1 and true amino acid availability (TAAA) of three protein diets was determined in experiment 2. 3. Results showed that, in the soybean meal and cottonseed meal, the AAAA and TAAA of most AA determined by the intact ganders were higher than in the caecectomised ganders; in the fish meal, the AAAA and TAAA of most AA determined by the intact ganders were lower than in the caecectomised ganders. 4. Results of the present study suggest that the effect of caecectomy on AAA in geese was dependent on the feedstuff assayed, and it was better to use caecectomised poultry for AAA assessment.

A comparison of methods to determine amino acid availability of feedstuffs in cecectomized ganders.

Experiments were conducted to compare endogenous amino acid losses and the true amino acid availability (TAAA) of 3 feedstuffs by using methods involving a short-term fasting and an N-free diet with cecectomized ganders. Diets were formulated to contain soybean meal, fish meal, and cottonseed meal as the sole source of protein. A precision-fed assay was used in which each feed sample was precise-fed (60 g) to geese and excreta were collected for 48 h. A N-free diet and fasting methods were used to evaluate the endogenous amino acid losses. Endogenous losses of 3 amino acids were significantly different (P < 0.01) with the N-free diet and fasting methods. The TAAA of soybean meal, fish meal, and cottonseed meal determined by N-free diet method ranged from 84.49 to 97.09%, 89.18 to 98.16%, and 77.09 to 98.32%, respectively. The TAAA of these 3 diets determined by the fasting method ranged from 83.50 to 97.77%, 88.08 to 99.60%, and 76.09 to 98.09%, respectively. However, there were only a few small differences (P > 0.05) between methods in each amino acid. In conclusion, there was no difference in determination of the amino acid availability of these feedstuffs using cecectomized ganders between the N-free diet and fasting methods.