Chun-qi Gao

Evaluation of the compositional and nutritional values of phytase transgenic corn to conventional corn in roosters

Three experiments were conducted to evaluate the compositional and nutritional values of corn grains [phytase transgenic corn (PTC) and isogenic conventional corn (CC)] and compare the efficacy of corn-based phytase and extraneous microbial phytase for enhancing the utilization of phytate phosphorus (P) in single corn or corn-soybean mixed meals (corn:soybean = 2.5:1, wt:wt) fed to roosters. Following a 48-h fasting period, 16 roosters were given 50 g of each sample via crop intubation and excreta were collected for 48 h. Nitrogen-free and phosphorus-free diets were used to evaluate endogenous amino acid and endogenous P losses, respectively. Chemical composition was not different between PTC and CC, whereas the phytase content for PTC was greater than CC (8,047 vs. 37 FTU/kg of corn, DM basis; P < 0.001). No difference was observed in the TME and true amino acid availability values between the PTC and CC in roosters. The true P utilization for PTC was greater than CC (37.92 vs. 55.85%; P < 0.001), and CC and PTC contained 0.13 and 0.19% available P (AP, DM basis; P < 0.001), respectively. There was no difference in P utilization (72.76 vs. 70.23%; P > 0.05) between roosters fed PTC and extraneous microbial phytase in equivalent FTU/kg of diets. The results of this study indicated that the chemical composition, TME, and true amino acid availability in PTC are essentially equivalent to that in CC, and the true P utilization for roosters is higher in PTC than in CC. Corn expressing phytase is as efficacious as equivalent microbial phytase when supplemented in corn-soybean diets for chickens.

Detection of Transgenic and Endogenous Plant DNA Fragments and Proteins in the Digesta, Blood, Tissues, and Eggs of Laying Hens Fed with Phytase Transgenic Corn

The trials were conducted to assess the effects of long-term feeding with phytase transgenic corn (PTC) to hens on laying performance and egg quality, and investigate the fate of transgenic DNA and protein in digesta, blood, tissues, and eggs. Fifty-week old laying hens (n = 144) were fed with a diet containing 62.4% PTC or non-transgenic isogenic control corn (CC) for 16 weeks. We observed that feeding PTC to laying hens had no adverse effect on laying performance or egg quality (P>0.05) except on yolk color (P<0.05). Transgenic phyA2 gene and protein were rapidly degraded in the digestive tract and were not detected in blood, heart, liver, spleen, kidney, breast muscle, and eggs of laying hens fed with diet containing PTC. It was concluded that performance of hens fed diets containing PTC, as measured by egg production and egg quality, was similar to that of hens fed diets formulated with CC. There was no evidence of phyA2 gene or protein translocation to the blood, tissues, and eggs of laying hens.

Growth of embryo and gene expression of nutrient transporters in the small intestine of the domestic pigeon (Columba livia)

The objective of this study was to investigate the relationship between gene expression of nutrient (amino acid, peptide, sodium and proton) transporters in the small intestine and embryonic growth in domestic pigeons (Columba livia). One hundred and twenty-five fertilized eggs were randomly assigned into five groups and were incubated under optimal conditions (temperature of 38.1 °C and relative humidity of 55%). Twenty embryos/birds from each group were sacrificed by cervical dislocation on embryonic day (E) 9, 11, 13, 15 and day of hatch (DOH). The eggs, embryos (without yolk sac), and organs (head, brain, heart, liver, lungs, kidney, gizzard, small intestine, legs, and thorax) were dissected, cleaned, and weighed. Small intestine samples were collected for RNA isolation. The mRNA abundance of intestinal nutrient transporters was evaluated by real-time reverse transcription-polymerase chain reaction (RT-PCR). We classified these ten organs into four types according to the changes in relative weight during embryonic development. In addition, the gene expression of nutrient transporters was differentially regulated by embryonic day. The mRNA abundances of b0,+AT, EAAT3, y+LAT2, PepT1, LAT4, NHE2, and NHE3 increased linearly with age, whereas mRNA abundances of CAT1, CAT2, LAT1, EAAT2, SNAT1, and SNAT2 were increased to higher levels on E9 or E11 and then decreased to lower levels until DOH. The results of correlation analysis showed that the gene expressions of b0,+AT, EAAT3, PepT1, LAT4, NHE2, NHE3, and y+LAT2 had positive correlations with body weight (0.71<correlation coefficient (CC)<0.82, P<0.0001), while CAT1, CAT2, EAAT2, SNAT1, and SNAT2 had negative correlations with body weight (−0.86<CC<−0.64, P<0.0001). The gene expressions of b0,+AT, EAAT3, LAT4, PepT1, NHE2, NHE3, and y+LAT2 showed positive correlations with intestinal weight (0.80<CC<0.91, P<0.0001),while CAT1, CAT2, and EAAT2 showed negative correlations with intestinal weight (−0.84<CC<−0.67, P<0.0001). It was concluded that the differences between growth trajectories of organs and gene expression of nutrient transporters in small intestine were due to their functional and physiological properties, which provided a comprehensive study of amino acid and peptide transporter mRNA in the small intestine during embryonic growth of pigeons.概要目的探索家鸽胚胎期器官生长发育及其小肠营养素转运载体基因表达规律, 为家鸽胚胎期的营养调控提供理论基础。创新点第一次较全面地研究了家鸽胚胎期小肠氨基酸转运载体基因表达规律。方法利用实时荧光定量逆转录聚合酶链反应 (real-timeRT-PCR) 技术, 同时结合了多内标校正方法对小肠营养素转运载体基因表达模式进行分析。结论在家鸽胚胎期发育的过程中, 不同的器官生长模式不完全一致。营养素转运载体b0,+AT、EAAT3、LAT4、PepT1、NHE2、NHE3 和y+LAT2 基因的表达与小肠重量呈正相关关系, CAT1、CAT2 和EAAT2 基因的表达与小肠重量呈负相关关系。

Degradation of zearalenone in swine feed and feed ingredients by Bacillus subtilis ANSB01G

Zearalenone (ZEA) and its derivatives are mycotoxins that can cause oestrogenic effects and impair the reproductive physiology of animals, especially in female swine. Strategies to reduce or eliminate ZEA contamination in foods and feeds are very much needed. Among 36 bacterial isolates obtained from a variety of animal intestinal chyme, mouldy foods and feeds, soils, etc., five isolates demonstrated the ability to reduce more than 50% of ZEA in a liquid medium; ANSB01G isolate taken from normal broiler intestinal chyme reduced ZEA the most, by 88.65%. Using physiological, biochemical, and 16S rRNA gene sequence analysis methods, the ANSB01G isolate was identified as Bacillus subtilis. Under simulated intestinal tract conditions, the ANSB01G B. subtilis isolate degraded 84.58, 66.34 and 83.04% of ZEA in naturally contaminated maize, dried distillers’ grains with solubles, and swine complete feed, respectively. The highest degradation of ZEA occurred when the mycotoxin was co-incubated with the whole bacte...

Effects of dietary α-lipoic acid, acetyl-l-carnitine, and sex on antioxidative ability, energy, and lipid metabolism in broilers

An experiment was conducted to evaluate the effects of dietary α-lipoic acid (LA), acetyl-l-carnitine (ALC), and sex on antioxidative ability, energy, and lipid metabolism in broilers. A total of 972 one-day-old broilers with equal sex were randomly assigned in a 3 × 3 × 2 factorial design using 3 LA, 3 ALC levels, and 2 sexes (6 replications, 9 birds/replication). The LA and ALC levels were 0, 50, and 100 mg/kg, respectively. Results showed that increased LA or ALC resulted in increased total antioxidant capacity and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and decreased levels of malondialdehyde in serum and liver of birds (P < 0.05). In addition, with increasing addition of LA or ALC, an increased (P < 0.01) level of insulin (Ins), as well as decreased (P < 0.05) levels of glucose and glucagon (Glu), were observed in serum of broilers. Total cholesterol and triglyceride (TG) levels decreased (P < 0.05) and nonesterified fatty acid, lipoprotein lipase, and lipase levels increased (P < 0.05) in serum with increased administration of LA or ALC. Moreover, a significant (P < 0.05) interaction of LA × ALC was observed for serum and liver SOD, serum GSH-Px, glucose, and TG levels. Birds fed diets containing 50 mg/kg of LA and 50 mg/kg of ALC had higher serum and liver SOD activities and lower serum glucose and TG levels than those fed diets containing 100 mg/kg of LA or ALC alone. The main effect of sex and all interactions among main effects (except LA × ALC) were not significant (P > 0.05) for all of the above parameters. Overall, the present data indicate that LA or ALC supplementation, or both, at low levels (50 or 100 mg/kg) improved antioxidative ability, energy metabolism, and lipid metabolism in broilers, and synergistic effects by the combined supplementation of LA and ALC were indicated by serum and liver SOD activities and serum glucose and TG levels.

Heat stress inhibits proliferation, promotes growth, and induces apoptosis in cultured Lantang swine skeletal muscle satellite cells

Proliferation suppression and apoptosis are the prominent characteristics induced by heat stress (HS) in cells, whereas the effects of HS on cell growth (mass accumulation) are unknown. In this study, Lantang swine (an indigenous breed of China) skeletal muscle satellite cells (SCs) were pre-cultured at 37 °C for 24 h. The HS group was subjected to HS at 41 °C, while the control group was maintained at 37 °C. Heat shock protein 70 (HSP70) expression and SC size are significantly increased (P<0.05) by HS, but cell proliferation is suppressed (P<0.05) and apoptosis is induced (P<0.05). HS led to a lower percentage of SCs in the G0/G1 phase (P<0.05) together with a higher percentage of SCs in the S phase (P<0.05). However, the percentage of SCs in the G2/M phase was decreased (P<0.05) at 48 h but then increased (P<0.05) at 72 h with HS. In addition, the phosphorylation ratios of protein kinase b (Akt), ribosomal protein S6 kinase (S6K), and ribosomal protein S6 were increased (P<0.05) by HS. Nevertheless, the phosphorylation ratios of the 4E binding protein 1 and the eukaryotic initiation factor-4E were indistinguishable (P>0.05) from those of the control group. The phosphorylation ratio of the mammalian target of rapamycin (mTOR) (Ser2448) increased(P<0.05) within 48 h, and apparent differences were abrogated at 72 h (P>0.05). Moreover, cleaved caspase-3 expression was increased at 72 h (P<0.05). These findings indicate that HS induces apoptosis and disrupts cell cycle distribution to decrease the number of cells. Additionally, HS can promote SC growth via an activated Akt/mTOR/S6K signaling pathway.概要目的探究热应激对蓝塘猪骨骼肌卫星细胞增殖与生长的影响, 分析其作用机制。创新点通过体外细胞模型研究了热应激对猪骨骼肌卫星细胞增殖与生长的影响,发现热应激通过 Akt/mTOR/S6K 途径调控细胞的生长。方法本试验选用1 日龄蓝塘猪背最长肌的骨骼肌卫星细胞, 对照组为37 °C 正常培养, 热应激组培养温度为41 °C, 其它培养条件相同。采用细胞计数、MTT 法、流式细胞术等手段分析比较热应激对蓝塘猪骨骼肌卫星细胞增殖与生长的影响; 利用实时逆转录聚合酶链式反应 (RT-PCR) 和蛋白质印迹法 (Western blot) 等方法, 检测了热休克蛋白HSP70、半胱氨酸蛋白酶-3 (caspase-3) 及哺乳动物雷帕霉素靶蛋白 (mTOR) 信号通路关键基因的表达。结论(1) 41 °C 热处理可引起蓝塘猪骨骼肌卫星细胞产生热应激反应; (2) 热应激72 h 会通过改变细胞周期、抑制细胞增殖并诱导细胞凋亡等共同作用减少细胞数目; (3) 热应激通过Akt/mTOR/S6K途径调控细胞的生长。

Flaxseed reduces the pro-carcinogenic micro-environment in the ovaries of normal hens by altering the PG and oestrogen pathways in a dose-dependent manner.

The objective of the present study was to find the optimum dose of flaxseed that would decrease PG and alter oestrogen pathway endpoints implicated in ovarian cancer. In the study, four groups of fifty 1.5-year-old chickens were fed different amounts of flaxseed (0, 5, 10 or 15% of their total diet) for 4 months and were then killed to collect blood and tissues. Levels of flaxseed lignan metabolites, Enterolactone (EL) and Enterodiol (ED) were measured in the serum, liver and ovaries by liquid chromatography-MS/MS, and n-3 and n-6 fatty acid (FA) levels were measured by GC. The effects of the varied flaxseed doses were assessed by measuring levels of PGE2 and oestrogen metabolites (16-hydroxyestrone (16-OHE1) and 2-hydroxyestrone (2-OHE1)) as well as by analysing the expression of the oestradiol metabolising enzymes CYP3A4 (cytochrome p450, family 3, subfamily A, polypeptide 4), CYP1B1 (cytochrome p450, family 1, subfamily B, polypeptide 1) and CYP1A1 (cytochrome p450, family 1, subfamily A, polypeptide 1) and that of oestrogen receptor α (ERα) in the ovaries. The ratio of n-3:n-FA increased with an increase in flaxseed supplementation and corresponded to a dose-dependent decrease in cyclo-oxygenase-2 protein and PGE2 levels. EL and ED increased in the serum, liver and ovaries with increased concentrations of flaxseed. Flaxseed decreased the expression of ERα in the ovaries. The ratio of 2-OHE1:16-OHE1 in the serum increased significantly in the 15% flaxseed diet, and there was a corresponding increase in CYP1A1 in the liver and decrease in CYP3A4 in the ovaries. CYP1B1 mRNA also decreased with flaxseed diet in the ovaries. The 15% flaxseed-supplemented diet significantly decreased inflammatory PGE2, ERα, CYP3A4, CYP1B1 and 16-OHE1, but it increased CYP1A1 and 2-OHE1, which thus reduced the inflammatory and pro-carcinogenic micro-environment of the ovaries.

Growth curves and age-related changes in carcass characteristics, organs, serum parameters, and intestinal transporter gene expression in domestic pigeon (Columba livia)

Two experiments were conducted to fit growth curves, and determine age-related changes in carcass characteristics, organs, serum biochemical parameters, and gene expression of intestinal nutrient transporters in domestic pigeon (Columba livia). In experiment 1, body weight (BW) of 30 pigeons was respectively determined at 1, 3, 7, 14, 21, 28, and 35 days old to fit growth curves and to describe the growth of pigeons. In experiment 2, eighty-four 1-day-old squabs were grouped by weight into 7 groups. On d 1, 3, 7, 14, 21, 28, and 35, twelve birds from each group were randomly selected for slaughter and post-slaughter analysis. The results showed that BW of pigeons increased rapidly from d 1 to d 28 (a 25.7-fold increase), and then had little change until d 35. The Logistic, Gompertz, and Von Bertalanffy functions can all be well fitted with the growth curve of domestic pigeons (R2>0.90) and the Gompertz model showed the highest R2value among the models (R2=0.9997). The equation of Gompertz model was Y=507.72×e-(3.76exp(-0.17t))(Y=BW of pigeon (g); t=time (day)). In addition, breast meat yield (%) increased with age throughout the experiment, whereas the leg meat yield (%) reached to the peak on d 14. Serum total protein, albumin, globulin, and glucose concentration were increased with age, whereas serum uric acid concentration was decreased (P<0.05). Furthermore, the gene expressions of nutrient transporters (y+LAT2, LAT1, B0AT1, PepT1, and NHE2) in jejunum of pigeon were increased with age. The results of correlation analysis showed the gene expressions of B0AT1, PepT1, and NHE2 had positive correlations with BW (0.73<correlation coefficient<0.80,P<0.0001). It can be concluded that, 28-day-old should be the latest market age for meat type pigeon. And the various physiological and functional properties of organs, serum profiles, and gene expression of nutrient transporters in small intestine might cause the differences in their development patterns.

Phytase transgenic corn in nutrition of laying hens: Residual phytase activity and phytate phosphorus content in the gastrointestinal tract

The residual activities of transgenic corn-derived and 2 commercial microbial phytases (PA and PB) along the gastrointestinal tract (GIT) of laying hens were compared to evaluate their relative resistance to hydrolysis in the GIT when added to P-deficient diets. The treatments consisted of a negative control (NC) diet containing 0.10 nonphytate P and an NC diet supplemented with transgenic corn-derived phytase (TCDP), PA, and PB at 500 to 5,000 FTU/kg of diet, respectively. Seven diets were fed to Hy-Line Brown laying hens (n = 504; 8 replicates of 9 hens per treatment) for 21 d. At the end of the experiment, the hens were killed and digesta samples from the crop, proventriculus and gizzard, jejunum, and ileum were collected and analyzed for residual phytase activities and phytate P content. Phytase activity in the transgenic corn was determined to be 8,980 FTU/kg of DM. The residual phytase activities along the GIT had increased (P < 0.01) with the addition of TCDP, PA, and PB to the NC diets. The TCDP had higher residual activity (P < 0.05) in the crop, proventriculus and gizzard, jejunum, and ileum as compared with the PA and PB activity. There was a decrease (P < 0.01) in the phytate P content of the digesta from all sources of phytase supplementation in the NC diets. Residual phytate P content decreased caudally along the GIT of hens. The results of this research indicate that phytase expressed in corn is as efficacious as the commercial microbial phytases (PA and PB) in P-deficient diets for the improvement of phytate P digestibility, which would eliminate the need for supplemental phytase and corn separately in laying hen diets.

Flaxseed and its components differentially affect estrogen targets in pre-neoplastic hen ovaries

Flaxseed has been studied for decades for its health benefits that include anti-cancer, cardio-protective, anti-diabetic, anti-inflammatory properties. The biologically active components that mediate these effects are the omega-3 fatty acids and the lignan, secoisolariciresinol diglucoside. We have previously shown that whole flaxseed supplemented diet decreases the severity and incidence of ovarian cancer while a 15% dose of flaxseed is most protective against inflammation and estrogen-induced chemical and genotoxicity. The objective of this study was to dissect the independent effects of the two flaxseed components on estrogen signaling and metabolism. Two and half year old hens were fed either a control diet, 15% whole flaxseed diet, defatted flax meal diet or 5% flax oil diet for 3 months after which the animals were sacrificed and blood and tissues were harvested. Whole flaxseed diet caused a decrease in expression of ERα. ERα target gene expression was assessed using RT(2) profiler PCR array. Some targets involved in the IGF/insulin signaling pathway (IRS1, IGFBP4, IGFBP5) were downregulated by flaxseed and its components. Flaxseed diet also downregulated AKT expression. A number of targets related to NF-kB signaling were altered by flaxseed diet including a series of targets implicated in cancer. Whole flaxseed diet also affected E2 metabolism by increasing CYP1A1 expression with a corresponding increase in the onco-protective E2 metabolite, 2-methoxyestradiol. The weak anti-estrogens, enterolactone, enterodiol and 2-methoxyestradiol, might be working synergistically to generate a protective effect on the ovaries from hens on whole flaxseed diet by altering the estrogen signaling and metabolism.