Agouti signalling protein (ASIP) gene: molecular cloning,sequence characterisation and tissue distribution in domestic goose

1. Agouti signalling protein (ASIP) is an endogenous antagonist of melanocortin-1 receptor (MC1R) and is involved in the regulation of pigmentation in mammals. The objective of this study was to identify and characterise the ASIP gene in domestic goose.

2. The goose ASIP cDNA consisted of a 44-nucleotide 5?-terminal untranslated region (UTR), a 390-nucleotide open-reading frame (ORF) and a 45-nucleotide 3?-UTR. The length of goose ASIP genomic DNA was 6176 bp, including three coding exons and two introns.

3. Bioinformatic analysis indicated that the ORF encodes a protein of 130 amino-acid residues with a molecular weight of 14.88 kDa and an isoelectric point of 9.73.

4. Multiple sequence alignments and phylogenetic analysis showed that the amino-acid sequence of ASIP was conserved in vertebrates, especially in the avian species.

5. RT-qPCR showed that the goose ASIP mRNA was differentially expressed in the pigment deposition tissues, including eye, foot, feather follicle, skin of the back, as well as in skin of the abdomen. The expression level of the ASIP gene in skin of the abdomen was higher than that in skin of the back.

6. Those findings will contribute to further understanding the functions of the ASIP gene in geese plumage colouring.

     

Growth performance,carcass traits and digestive function of broiler chickens fed diets with graded levels of corn resistant starch

ABSTRACT

1. This study was conducted to assess the effects of graded levels of dietary corn resistant starch (RS) on growth performance, carcass traits, nutrient retention, digestive organ index, intestinal morphology, digestive enzyme activities, and mRNA expression of certain nutrient transporters in broiler chickens.

2. A total of 320, 1-d-old Arbor Acres broiler chickens were randomly assigned to five dietary treatments, with eight replicates of eight birds in each. These treatments included one corn-soybean control diet, a corn-soybean based diet containing 20% corn starch, and three diets supplemented with 4%, 8% and 12% RS by replacing corn starch with 6.67%, 13.33% and 20% of Hi-Maize 260® (identified as control, RS1, RS2, RS3 and RS4, respectively). The feeding period lasted 42 days.

3. Performance parameters including feed consumption, feed conversion, body weight gain and percentage of abdominal fat at d 42 of age, nutrient retention (including dry matter, fat, total starch and nitrogen free extract), and apparent metabolisable energy was measured from d 18 to 20 and d 39 to 41 and showed negative linear responses to increasing dietary RS level (P < 0.05). Birds fed the RS3 and RS4 diets showed higher relative weight of duodenum, jejunum and ileum, as well as lower villus height and villus height/crypt depth compared to the control (P < 0.05). The activity of pancreatic trypsin of birds at d 21 and 42 of age decreased linearly in response to the increase of dietary RS level (P < 0.01). There were linear changes in up-regulated mRNA expression of SGLT-1 and down-regulated mRNA expression of GLUT-2 with increasing proportion of RS at d 21 and 42 of age (P < 0.05), respectively.

4. It was concluded that feeding broilers with diets containing higher concentrations of RS impaired the development of small intestine, which resulted in lower apparent total tract retention of nutrients and poorer body weight gain, feed efficiency and carcass traits of broiler chickens.     

Paired box 7 (Pax7) gene: molecular characterisation,polymorphism and its association with growth performance in goose (Anser cygnoides)

1. Paired box (Pax7) gene is a member of the paired box family and plays a critical role in animal growth and muscle development. However, the molecular characterisation of the goose Pax7 gene is unknown.

2. The open-reading frame of goose Pax7 is composed of 1509 bp, which encodes a protein of 503 amino acids and shares high homology with Pax7 of other birds.

3. Ten single-nucleotide polymorphisms were identified in the genomic DNA sequence, 8 located in the intron region and two located in the exon region.

4. Association analysis showed the C122T locus was significantly associated with the body weight of Zhedong-White geese in week 4, 6, 8, 10 and 12.

5. It was concluded that the goose Pax7 gene may be an important candidate gene for goose growth traits and marker-assisted selection.     

Molecular cloning and expression analysis of pig CD7

CD7 is an integral membrane protein which mediates an important signal to mediate the differentiation, activation, and regulation of some T cells and NK cells. However, only human and mouse CD7 have been identified and studied among mammalian species. In this study, we cloned pig CD7 cDNA and determined its complete cDNA sequence. Pig CD7 cDNA contained an open reading frame (627 bp) encoding 208 amino acids with well conserved motifs involved in signal transduction within cytoplasmic tail among mammalian species. Pig CD7 mRNA was detected by RT-PCR in mainly lymphoid tissues, indicating the conserved functions of CD7 in pigs. Moreover, we generated soluble pig CD7 fusion immunoglobulin (pig CD7Ig) containing extracellular domain of pig CD7 to test whether pig CD7 binds to pig galectin-3. Flow cytometry and immunohistochemistry analyses indicated that soluble pig CD7Ig can bind to galectin-3 expressed in macrophages and epithelial cells of small intestine. These results help to analyze the structural relationship between CD7 and its ligand transferring signal transduction among mammalian species.     

Effects of full-fat rice bran inclusion in diets on growth performance and meat quality of Sichuan goose

1. This trial was conducted to study the effect of full-fat rice bran inclusion in diets on growth performance, carcass and meat quality and fatty acid composition in Sichuan goose.

2. A total of 204 Sichuan white male geese (28-d-old, 984 ± 15 g) were used in the 42-d assay. Full-fat rice bran inclusion in diets was 0%, 6%, 12% and 18%, respectively.

3. On d 70, two geese from each pen were randomly selected and killed for measuring the carcass and meat quality and the meat fatty acid composition.

4. The results showed that full-fat rice bran inclusion had no effect on average daily gain during 28–56 d, but rice bran inclusion at 18% increased average daily gain during 57–70 d. In addition, the full-fat rice bran supplementation decreased the subcutaneous fat yield, and the inclusion of full-fat rice bran in amounts of 12% and 18% decreased the half-eviscerated carcass yield, eviscerated carcass yield and crude fat content in goose meat. Moreover, full-fat rice bran supplementation had no effect on the content of total saturated fatty acid (SFA), but decreased the content of total monounsaturated fatty acid (MUFA). The inclusion of full-fat rice bran in amounts of 12–18% increased the content of total polyunsaturated fatty acid (PUFA) and total n-6 in goose meat and in the amount of 18% increased n-3 fatty acids content in goose meat.

5. The results indicated that the rice bran inclusion had a positive effect in geese by stimulating growth performance and improving meat quality and fatty acid composition of goose meat.

     

鸭脂肪酸结合蛋白基因的克隆和序列多态性分析

根据鸡脂肪型脂肪酸结合蛋白（A-FABP）基因序列设计1对引物对鸭基因组进行PCR扩增，将扩增出的475bp片段进行克隆和测序，序列分析表明该基因片段包含部分外显子2、3和完整的内含子2序列，与鸡、鹅、猪的A-FABP基因分别有84％、94％和77％的同源性。在此基础上又设计3对引物利用PCR—SSCP方法对获得的序列进行多态性研究。结果引物P3在樱桃谷鸭、山麻鸭等品种中发现4处单碱基突变，分别为：241处“A～T”突变；243处的“T—C”突变；258处的“T—C”突变；299处的“C—T”突变。这些点突变共产生了3种基因型，且基因型分布在各鸭品种间有显著差异。     

Molecular cloning,alternative splicing and mRNA expression analysis of MAGI1 and its correlation with laying performance in geese

1. In a previous study, a laying-related single nucleotide polymorphism (SNP) Record-106975 (A–G) was found in the intron of Membrane-associated Guanylate Kinase with Inverted Domain Structure-1 (MAGI1) gene by restriction site-associated DNA sequencing. This study was performed to further identify the function of MAGI1 on laying performance in Yangzhou geese.

2. The complete MAGI1 cDNA sequence was cloned from the ovary of the Yangzhou goose. A total of 4 alternative splice variants were identified, and the characterisation of their nucleic acid and amino acid (AA) sequences were analysed. The AA sequence of goose MAGI1 was highly conserved with other avian species.

3. MAGI1 was ubiquitously expressed in 11 tested tissues, with greater amounts of mRNA in the granulosa cell layer and ovary. GG (n = 14) and GA (n = 6) genotypes of SNP Record-106975 showed significantly lower MAGI1 mRNA levels than those of the AA genotype.

4. Over-expression and knockdown experiments revealed that MAGI1 mRNA expression was associated with caspase-3 mRNA levels in granulosa cells, and may play a role in regulating apoptosis of granulosa cells.     

MyoG和A—FABP基因多态性与鹅肉品质性状的相关性研究

为了检测MyoG基因和A-FABP基因单核苷酸多态性（SNP）与鹅肉品质性状的相关性,以太湖鹅为研究对象,测定肌内脂肪含量和相关组织学指标特性,利用PCR—SSCP和测序技术研究MyoG基因外显子1和A—FABP基因内含子2SNPs,并进行相关分析。研究结果表明：（1）MyoG基因外显子1扩增序列大小为420bp,共有1个突变,突变位点在64bp处,为C—T突变,不同基因型之间肌纤维密度、直径差异显著（P〈0．05）;（2）A—FABP基因扩增序列大小为482bp,共发现3个突变位点,在第187bp处发生C—T突变,在第235bp处发生C—A突变,在第372bp处发生A—C突变;（3）A—FABP基因内含子2扩增产物经SSCP分析后发现两种基因型,不同基因型之间脂肪含量有一定差异。比对发现其与家鸭同源性达91％,与家鸡的同源性为85％。本研究结果为进一步分析MyoG和A—FABP基因的遗传变异及其与肉质性状的相关性以及将该分子标记应用于育种计划奠定一定的理论基础。     

Glucose participates in the formation of goose fatty liver by regulating the expression of miRNA-33/CROT

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.     

The effect of heat stress on GHR,IGF-I,ANT, UCP and COXIII mRNA expression in the liver and muscle of high and low feed efficiency female quail

1. A study was conducted to test the hypothesis that feed efficiency (FE) correlated with the expression of genes from the somatotropic axis and mitochondrial genes involved in energy production, and that the environment to which the birds are exposed influenced the expression of such genes.

2. Quails were divided into High-FE and low-FE groups and maintained in a comfortable or heat stress (HS) (38°C for 24 h) environment to evaluate changes in insulin-like growth factor I (IGF-I), growth hormone receptor (GHR), adenine nucleotide translocator (ANT), uncoupling protein (UCP) and cytochrome oxidase subunit III (COX III) messenger ribonucleic acid (mRNA) expression in liver and muscle tissues.

3. High-FE quails (0.28 g/g) presented a higher final body weight, greater weight gain and a better feed conversion ratio than low-FE birds (0.18 g/g). High-FE birds showed greater IGF-I mRNA expression in the liver and muscle and greater GHR mRNA expression in the muscle.

4. Environmental effects only affected GHR expression in the liver, with quails under comfortable conditions exhibiting greater GHR expression than quails subjected to HS.

5. There was a significant interaction between FE and environmental temperature on ANT mRNA expression in the liver. The greatest ANT mRNA expression was observed for high FE-birds that remained under comfortable conditions.

6. In the liver, UCP mRNA expression did not differ among the quails and was not affected by environment or efficiency. However, comparisons of the low- and high-FE birds revealed higher levels of UCP mRNA in the muscle of low-FE birds.

7. COX III mRNA expression in the liver was dependent on environmental temperature and FE. Higher COX III mRNA expression was observed in animals that remained under comfortable conditions, and high-FE birds exhibited higher expression levels compared to low-FE birds.

8. These results suggest a correlation between IGF-I, GHR, ANT, UCP and COX III gene expression and FE and that environmental temperature could affect the expression of some of these genes.     

Molecular cloning,expression and regulation analysis of the interleukin-6 (IL-6) gene in goose adipocytes

1. Interleukin-6 (IL-6) is a multifunctional cytokine involved in lipid metabolism in adipose tissue. The objective of the study was to identify and characterize the IL-6 gene in the goose.

2. A full-length coding sequence (CDS) of the goose (Anser anser) IL-6 gene was cloned that encoded a 234-amino acid peptide containing a 38-amino acid signal peptide, an IL-6/G-CSF/MGF family consensus pattern and four conserved α-helices. The mature goose IL-6 showed 74% and 39% identities to that of chicken and human, respectively.

3. Quantitative real-time PCR analysis showed that the goose IL-6 was predominantly expressed in liver and was up-regulated in adipocytes by lipopolysaccharide (LPS) and oleic acid.     

Effect of dietary fibre and grit on performance,gastrointestinal tract development,and grit pattern of goose

ABSTRACT

1. This study aimed to investigate effects of dietary fibre and grit on growth performance, gastrointestinal tract development, and gizzard grit retention of geese.

2. The trial had a 3 × 2 factorial design consisting of three levels of dietary crude fibre (CF, 4%, 7% and 10%, adjusted by grass powder), with or without grit addition (1–4 mm river sand).

3. In total, 648, 22-d-old male goslings were randomly allotted to six treatments (six pens/treatment). At 49 d and 70 d of age, one goose per pen was euthanised to collect samples.

4. The birds fed 10% CF had decreased feed conversion ratio (FCR) during 22–49 d, but this effect was less in older geese. Increasing dietary CF levels increased relative weights of gizzards for geese aged 49 d and 70 d. Grit addition decreased the relative weights of gizzard and duodenum of geese aged 49 d. The gizzard of geese could selectively retain grit from feed even with no grit added. With adequate supply, most grit in gizzard was 0.45–3 mm in size.

5. In conclusion, supplement of CF and grit mainly affected gastrointestinal tract, and the amount of CF affected FCR. Geese aged 22–49 d could utilise dietary CF levels of 4%-7% and older birds could feed on diets with up to 10% CF. The gizzard of goose selectively retained grit of a particle size of 0.45–3 mm.     

Effects of pea extrusion and enzyme and probiotic supplementation on performance,microbiota activity and biofilm formation in the broiler gastrointestinal tract

ABSTRACT

1. The effects of supplementation of broiler chicken diets with pea meal, carbohydrase enzymes and a probiotic were investigated for potential performance improvement.

2. Raw or extruded pea meal (cv Model, grown in Poland) was included in a wheat-soybean meal-based diet at 250 g/kg. The diets were unsupplemented (control) or supplemented with either carbohydrase enzymes (200 U/kg xylanase and 10 U/kg β-glucanase in feed) or a probiotic (Bacillus subtilis), or both. The diets were fed to Ross 308 broilers aged 9–28 days.

3. After two additional days, chick gastrointestinal tracts were excised and analysed for the presence of Bacillus subtilis biofilm; and the ileal and caecal digesta were analysed for bacterial enzyme activities and to determine the concentration of short-chain fatty acids (SCFAs).

4. Feeding the pea-based diet supplemented with the probiotic compromised feed utilisation, due to higher feed intake. The addition of enzymes to the raw, but not the extruded, pea containing diet partially ameliorated this effect (pea form × additives; P < 0.002).

5. In the ileal digesta, interactions between the dietary treatments were observed for the activities of all bacterial glycolytic enzymes and for SCFA concentrations. β-glucosidase, α-galactosidase and β-glucuronidase were highest in birds fed the diet containing extruded pea supplemented with the probiotic and enzymes (pea form x additives; P = 0.018 to P < 0.006). In the caecal digesta, interactions were observed for bacterial enzyme activities, but not for total SCFA concentration. Biofilm formation in the caecum indicated that the probiotic strain was metabolically active in the broiler gut.

6. In conclusion, supplementation of diets containing raw or extruded pea meal with enzymes and a Bacillus subtilis spore-based probiotic modulated microbiota activity but had no clear effects on broiler performance. Probiotic administration did not cause excessive fermentation in the ileum and caecum but enhanced Bacillus subtilis spp. biofilm formation in the caecum, which may be indicative of a beneficial effect on gut health.     

溆浦鹅肉营养成分与品质分析

参照国家食品相关标准分析的测定方法对溆浦鹅胸肌和腿肌肉营养成分和肉品质进行了比较分析和营养品质评价。结果表明:溆浦鹅屠宰率、半净膛率、全净膛率分别为90.11%、80.45%、68.62%,脂肪率为4.93%,肝重100.75 g;屠宰24 h后pH在5.71~5.94之间,胸肌肌肉失水率27.54%,腿肌失水率20.58%,粗蛋白、粗脂肪、粗灰分、水分分别为22.64%、2.72%、2.30%、70.17%;肉中铁、铜、锌、镁、钙、硒平均含量分别为6.97 mg/100g、0.41 mg/100g、2.57 mg/100g、39.94 mg/100g、13.84 mg/100g、18.71μg/100g;肌纤维直径39.52μm,肌纤维密度630.1根/mm2;17种氨基酸总量(干样)为568.53 mg/g,必需氨基酸、甜味氨基酸、鲜味氨基酸平均比例分别为46.74%、26.12%、34.22%,根据氨基酸评分,溆浦鹅的第一限制性氨基酸为蛋氨酸+胱氨酸,第二限制性氨基酸为缬氨酸;肌苷酸1.435 mg/g;饱和脂肪酸32.45%,单不饱和脂肪酸46.6%,多不饱和脂肪酸21.24%,胆固醇55.94 mg/100g。与常规禽肉对比,溆浦鹅肉蛋白高、脂肪低、产肉性能好,含较丰富的硒、钙、镁、铁等矿物元素,且胆固醇含量低、肌苷酸含量高,有较好的食用价值与保健作用。     

Haematological parameters,serum lipid profile,liver function and fatty acid profile of broiler chickens fed on diets supplemented with pomegranate seed oil and linseed oil

1. The objective of the present study was to determine effect of pomegranate seed oil (PSO) and linseed oil (LO) on haematological parameters, serum lipid profile and liver enzymes as well as fatty acids profile of adipose tissue in broilers.

2. Broilers (n = 400) were fed on diets containing graded PSO levels (0.0%, 0.5%, 1.0%, 1.5%) with or without 2% LO. After 6 weeks of feeding, 6 male broilers from each group were slaughtered and abdominal fat, liver and blood samples were collected.

3. Mixtures of pomegranate seed oil (0.5%, 1%) with linseed oil increased white blood cell level in broilers. Total cholesterol was elevated after LO supplementation whereas administration of PSO (1.5%) significantly decreased this parameter. PSO administration caused c9,t11 conjugated linoleic acid (CLA) concentration-dependent deposition in adipose tissue. By LO addition α-linolenic acid (ALA) content was enhanced, decreasing the n-6/n-3 ratio. PSO and ALA also affected oleic acid proportion in adipose tissue. Neither pomegranate seed oil nor linseed oil had any effect on liver parameters.

4. Pomegranate seed oil had no negative effects on broiler health status and can be considered as a functional poultry meat component.

     

Dietary conjugated linoleic acid affects blood parameters,liver morphology and expression of selected hepatic genes in laying hens

1. The objective of this research were to investigate the effect of a conjugated linoleic acid (CLA)-enriched diet on Isa Brown laying hen health status and to provide a comprehensive analysis of changes in blood parameters, liver morphology and selected hepatic gene expression.

2. Hens were allocated to the control and experimental group (diet enriched with 0.75% CLA) for a total period of 4 m. At the end of the experiment half of the hens from each group were slaughtered for analyses. The remaining hens were transferred to an organic farm for the next 5 m and fed on the diet without CLA supplementation.

3. The CLA-enriched diet resulted in significant changes in blood and serum parameters; specifically, haematocrit (HCT), mean corpuscular volume (MCV) and white blood cells (WBC) count were decreased compared to the control. The total cholesterol (TC) was not significantly affected while the triacylglycerol’s (TG) concentration was elevated. The activity of alanine aminotransferase (ALT) was significantly increased in the CLA-supplemented group, while aspartate aminotransferase (AST) showed an increasing tendency. Liver biopsies showed pathological changes classified as non-alcoholic fatty liver disease (NAFLD). Additionally, the expression of hepatic genes involved in fatty acids synthesis (ME1, ACLY, ACC, FASN, SCD1), oxidation (CPT1α, PPARA), detoxification processes (Cytochrome P450, CYP, Flavin-containing monooxygenase, FMO3), oxidative stress (NOX4, XbP1) and inflammation (IL6, TNFα) were elevated. Cessation of CLA supplementation for 5 m of organic farming resulted in normalisation of blood and hepatic parameters to the levels observed in control hens.

4. The results of this study indicate that dietary CLA triggers an integrated stress response in laying hens and activates mechanisms involved in liver detoxification.