Rapid Identification and Characterization of Antioxidants from Ligularia fischeri

Xiang-Lan Piao,Xiao-Yuan Mi,Yan-Ze Tian,Qian Wu,Hui-Shan Piao,Zhikai Zeng,Xiangshu Piao,Ding Wang 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.12

The objectives of this study were to investigate the radical-scavenging activity of Ligularia fischeri on oxidative damage by the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and to rapidly identify the active components using the bioassay-linked fractionation method. The MeOH extract and fractions of CH2Cl2, BuOH, and H2O from L. fischeri showed DPPH radical-scavenging effects in a dose-dependent manner (p < 0.01). In particular, the BuOH fraction had the most effective (p < 0.05) antioxidative capacity. The active constituents from the BuOH fraction of L. fischeri were rapidly isolated by bioassay-linked HPLC method and identified as hyperoside and 2''-acetylhyperoside with potent antioxidant effects against the DPPH radical, with IC50 values of 1.31 and 7.09 μg/mL, respectively. They have not been reported from L. fischeri yet.

Effect of Bacillus amyloliquefaciens-based Direct-fed Microbial on Performance, Nutrient Utilization, Intestinal Morphology and Cecal Microflora in Broiler Chickens

Lei, Xinjian,Piao, Xiangshu,Ru, Yingjun,Zhang, Hongyu,Peron, Alexandre,Zhang, Huifang Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.2

The present study was conducted to evaluate the effect of the dietary supplementation of Bacillus amyloliquefaciens-based direct-fed microbial (DFM) on growth performance, nutrient utilization, intestinal morphology and cecal microflora in broiler chickens. A total of two hundred and eighty eight 1-d-old Arbor Acres male broilers were randomly allocated to one of four experimental treatments in a completely randomized design. Each treatment was fed to eight replicate cages, with nine birds per cage. Dietary treatments were composed of an antibiotic-free basal diet (control), and the basal diet supplemented with either 15 mg/kg of virginiamycin as antibiotic growth promoter (AGP), 30 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 30) or 60 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 60). Experimental diets were fed in two phases: starter (d 1 to 21) and finisher (d 22 to 42). Growth performance, nutrient utilization, morphological parameters of the small intestine and cecal microbial populations were measured at the end of the starter (d 21) and finisher (d 42) phases. During the starter phase, DFM and virginiamycin supplementation improved the feed conversion ratio (FCR; p<0.01) compared with the control group. For the finisher phase and the overall experiment (d 1 to 42) broilers fed diets with the DFM had better body weight gain (BWG) and FCR than that of control (p<0.05). Supplementation of virginiamycin and DFM significantly increased the total tract apparent digestibility of crude protein (CP), dry matter (DM) and gross energy during both starter and finisher phases (p<0.05) compared with the control group. On d 21, villus height, crypt depth and villus height to crypt depth ratio of duodenum, jejunum, and ileum were significantly increased for the birds fed with the DFM diets as compared with the control group (p<0.05). The DFM 30, DFM 60, and AGP groups decreased the Escherichia coli population in cecum at d 21 and d 42 compared with control group (p<0.01). In addition, the population of Lactobacillus was increased in DFM 30 and DFM 60 groups as compared with control and AGP groups (p<0.01). It can be concluded that Bacillus amyloliquefaciens-based DFM could be an alternative to the use of AGPs in broilers diets based on plant protein.

Effects of Adding Essential Oil to the Diet of Weaned Pigs on Performance, Nutrient Utilization, Immune Response and Intestinal Health

Li, Pengfei,Piao, Xiangshu,Ru, Yingjun,Han, Xu,Xue, Lingfeng,Zhang, Hongyu Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.11

The objective of this study was to evaluate the effects of adding essential oils to the diet of weaned pigs on performance, nutrient utilization, immune response and intestinal health. A total of 96 weaning pigs ($8.37{\pm}1.58$ kg) were allotted to one of three dietary treatments. The treatments consisted of an unsupplemented basal diet (negative control, NC) or similar diets supplemented with 0.01% of an essential oil product which contained 18% thymol and cinnamaldehyde (EOD) as well as a diet supplemented with 0.19% of an antibiotic mixture which provided 150 ppm chlortetracycline, 80 ppm colistin sulfate and 50 ppm kitasamycin (positive control, PC). Each treatment was provided to eight pens of pigs with four pigs per pen. Over the entire 35 d experiment, ADG and fecal score were improved (p<0.05) for pigs fed the PC and EOD compared with the NC. Dry matter and crude protein digestibility as well as lymphocyte proliferation for pigs fed the PC and EOD diets were increased significantly compared with NC (p<0.05). IGF-I levels in plasma were significantly increased (p<0.05) in pigs fed the PC diet compared with pigs fed the NC diet. Interleukin-6 concentration was lower (p<0.05) and the tumor necrosis factor-${\alpha}$ level was higher (p<0.05) in the plasma of pigs fed the EOD diet than the NC diet. Plasma total antioxidant capacity level increased (p<0.05) in pigs fed the EOD diet compared with pigs fed the NC. Villus height to crypt depth ratio in the jejunum was greater (p<0.05) in pigs fed the PC and EOD diets than the NC. The numbers of E. coli in the cecum, colon and rectum were reduced (p<0.05) in pigs fed the PC and EOD diets compared with the control. In the colon, the ratio of Lactobacilli to E. coli was increased (p<0.05) in pigs fed the EOD diet compared with NC diet. Total aerobe numbers in the rectum were decreased (p<0.05) in pigs fed the PC and EOD diets compared with the control. Collectively, these results indicate that blends of essential oils could be a candidate for use as an alternative to traditional antibiotics in weaning pig diets.

The Optimum Methionine to Methionine Plus Cystine Ratio for Growing Pigs Determined Using Plasma Urea Nitrogen and Nitrogen Balance

Qiao, Shiyan,Piao, Xiangshu,Feng, Zhanyu,Ding, Yuhua,Yue, Longyao,Thacker, P.A. Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.3

The objective of this study was to determine the optimum ratio of methionine to methionine plus cystine for growing pigs. A nitrogen balance trial was conducted using a total of 21 barrows (Large WhiteLandrace) over two replicates. The initial body weight was $20.36{\pm}1.22kg$ (mean${\pm}$SD) in the first replicate and $23.54{\pm}1.02kg$ (mean${\pm}$SD) in the second. For each replicate, the 21 pigs were randomly assigned to one of seven dietary treatments with three observations per treatment. The diets included a methionine and cystine-deficient basal diet with all other essential nutrients meeting nutrient requirements and six diets formulated with graded levels of DL-methionine (0.00, 0.03, 0.06, 0.10, 0.13, 0.16%) and $L-Cystine{\cdot}HCl{\cdot}H_2O$ (0.19, 0.15, 0.11, 0.07, 0.04, 0.00%). This resulted in ratios of methionine to methionine plus cystine of 41.3, 29.6, 35.3, 41.2, 46.0, 51.6 and 57.5%. Each experimental period lasted 12 days consisting of a seven-day adaptation period followed by a five-day total collection of urine and feces. During the collection period, pigs were fed 900 g/day for the first replicate and 1,200 g/day for the second replicate. The feed was provided in three equal portions at 0800, 1500, and 2200 h daily. Pigs had ad libitum access to water after feeding. There was a linear (p<0.01) and quadratic (p<0.01) effect on daily gain and feed conversion as the ratio of methionine to methionine plus cystine increased. Pigs receiving the diets providing a methionine to methionine plus cystine ratio of 51.6% had the best daily gain and feed conversion. Plasma urea nitrogen was also lowest for this treatment. Nitrogen retention increased (p<0.01) as the relative proportion of methionine increased up to 51.6% and then a downward trend occurred at 57.5%. The quadratic regression model, as well as one- and two- slope regression line models, were used to determine the optimum ratio of methionine to methionine plus cystine. Eliminating the 35.3% methionine to methionine plus cystine treatment resulted in $R^2$ values in excess of 0.92. The optimal ratio of methionine to methionine plus cystine was estimated to be 54.15% for nitrogen retention and 56.72% for plasma urea nitrogen.

Effect of Bacillus amyloliquefaciens-based Direct-fed Microbial on Performance, Nutrient Utilization, Intestinal Morphology and Cecal Microflora in Broiler Chickens

Xinjian Lei,Xiangshu Piao,Yingjun Ru,Hongyu Zhang,Alexandre Peron,Huifang Zhang 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.2

The present study was conducted to evaluate the effect of the dietary supplementation of Bacillus amyloliquefaciens-based direct-fed microbial (DFM) on growth performance, nutrient utilization, intestinal morphology and cecal microflora in broiler chickens. A total of two hundred and eighty eight 1-d-old Arbor Acres male broilers were randomly allocated to one of four experimental treatments in a completely randomized design. Each treatment was fed to eight replicate cages, with nine birds per cage. Dietary treatments were composed of an antibiotic-free basal diet (control), and the basal diet supplemented with either 15 mg/kg of virginiamycin as antibiotic growth promoter (AGP), 30 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 30) or 60 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 60). Experimental diets were fed in two phases: starter (d 1 to 21) and finisher (d 22 to 42). Growth performance, nutrient utilization, morphological parameters of the small intestine and cecal microbial populations were measured at the end of the starter (d 21) and finisher (d 42) phases. During the starter phase, DFM and virginiamycin supplementation improved the feed conversion ratio (FCR; p<0.01) compared with the control group. For the finisher phase and the overall experiment (d 1 to 42) broilers fed diets with the DFM had better body weight gain (BWG) and FCR than that of control (p<0.05). Supplementation of virginiamycin and DFM significantly increased the total tract apparent digestibility of crude protein (CP), dry matter (DM) and gross energy during both starter and finisher phases (p<0.05) compared with the control group. On d 21, villus height, crypt depth and villus height to crypt depth ratio of duodenum, jejunum, and ileum were significantly increased for the birds fed with the DFM diets as compared with the control group (p<0.05). The DFM 30, DFM 60, and AGP groups decreased the Escherichia coli population in cecum at d 21 and d 42 compared with control group (p<0.01). In addition, the population of Lactobacillus was increased in DFM 30 and DFM 60 groups as compared with control and AGP groups (p<0.01). It can be concluded that Bacillus amyloliquefaciens-based DFM could be an alternative to the use of AGPs in broilers diets based on plant protein.

The Apparent Digestibility of Corn By-products for Growing-finishing Pigs In vivo and In vitro

Guo, Liang,Piao, Xiangshu,Li, Defa,Li, Songyu Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.3

Two trials in vivo and in vitro were conducted, in vivo to determine the apparent digestibility of gross energy, crude protein, dry matter, acid detergent fiber, neutral detergent fiber and apparent digestible energy in 10 corn by-products. In vivo the diets included one basal corn diet, four corn gluten meal diets, four corn distillers dried grains with solubles diets and two corn distillers dried grains diets using the different methods, 12 crossbred barrows weigh $40{\pm}$1.6 kg were allocated into individual metabolic crate, according to a $6{\times}6$ Latin square design. In vitro using flask technique, filter bag technique and dialysis tubing technique, the digestibilities of gross energy, crude protein and dry matter in corn gluten meal and corn distillers dried grains with solubles were investigated. Pepsin, pancreatin, intestinal fluid, rumen fluid and cellulase were used in incubation. The results showed that correlation coefficient was 0.73 in corn distillers dried grains with solubles between the digestibility of crude protein and acid detergent fiber in vivo (p<0.01); and correlation coefficient was 0.68 in corn distillers dried grains with solubles between the digestibility of gross energy and neutral detergent fiber in vivo (p<0.01). Apparent digestible energy (DE) of corn by-products in pig total tract was predicted by the percentage of crude protein (CP) and the content of gross energy (GE) in feedstuff. The equation: DE=5,601.09+26.69$\times$CP %-0.5904$\times$GE, ($R^2=0.72$). In vitro, filter bag technique was more convenient; furthermore, the digestibility for the treatments (pepsin+pancreatin+rumen fluid and pepsin+pancreatin+cellulase) was better.

Effects of Lactobacilli on the Performance, Diarrhea Incidence, VFA Concentration and Gastrointestinal Microbial Flora of Weaning Pigs

Huang, Canghai,Qiao, Shiyan,Li, Defa,Piao, Xiangshu,Ren, Jiping Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.3

Two experiments were conducted to evaluate the effects of a complex Lactobacilli preparation on performance, resistance to E. coli infection and gut microbial flora of weaning pigs. In exp. 1, twelve pigs (7.65$\pm$1.10 kg BW), weaned at 28 d, were randomly allotted into 2 groups and placed in individual metabolic cages. During the first 7 d, one group of pigs was provided ad libitum access to water containing $10^5$ colony forming units (CFU) Lactobacilli per ml and the control group was provided tap water. The Lactobacilli preparation included Lactobacillus gasseri, L. reuteri, L. acidophilus and L. fermentum, which were isolated from the gastrointestinal (GI) tract mucosa of weaning pigs. On d 8, 20 ml of $10^8$ CFU/ml E. coli solution (serovars K99, K88 and 987P at the ratio of 1:1:1) was orally administered to each pig. Diarrhea scores and diarrhea incidence were recorded from d 7 to 14. On d 14, pigs were euthanized and digesta and mucosa from the stomach, duodenum, jejunum, ileum, cecum and colon were sampled using aseptic technique to determine microflora by culturing bacteria in selective medium. The results showed that Lactobacilli treatment significantly decreased E. coli and aerobe counts (p<0.01) but increased Lactobacilli and anaerobe counts (p<0.01) in digesta and mucosa of most sections of the GI tract. A 66 and 69.1% decrease in diarrhea index and diarrhea incidence, respectively, was observed in the Lactobacilli treated group. In exp. 2, Thirty-six crossbred Duroc$\times$Landrace$\times$Yorkshire piglets, weaned at 28$\pm$2 days, were selected and randomly allocated into 2 groups. There were 18 piglets in each group, 3 piglets in one pen and 6 replicates in each treatment with 3 pens of barrow and 3 pens of female piglet in each treatment. Piglets had ad libitum access to feed and water. The initial body weight of piglet was 7.65$\pm$1.09 kg. Dietary treatments included a non-medicated basal diet with Lactobacilli ($10^5$ CFU/g diet) or carbadox (60 mg/kg) as control. On d 21, six pigs per group (one pig per pen) were euthanized. Ileal digesta was collected to determine apparent amino acid digestibility. Microflora content was determined similarly to exp.1. The results showed that Lactobacilli treatment significantly improved average daily feed intake (ADFI) of pigs compared to carbadox (p<0.05) during the first 2 wks after weaning and average daily gain (ADG) and ADFI increased significantly (p<0.05) from d 8 to 14. Nitrogen and total phosphorus digestibility also increased (p<0.05). Bacterial counts were similar to exp. 1. The results indicate that the complex Lactobacilli preparation improved performance for 2 wks after weaning, enhanced resistance to E. coli infection, and improved microbial balance in the GI tract.

Effects of proteinate complex zinc on growth performance, hepatic and splenic trace elements concentrations, antioxidative function and immune functions in weaned piglets

Yue She,Qiang Huang,Defa Li,Xiangshu Piao 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.8

Objective: To assess the effects of proteinate complex zinc (PC-Zn) on growth performance, antioxidative function, trace element concentrations and immune function in weaned piglets. Methods: Three hundred newly weaned barrows (Duroc×Landrace×Yorkshire), 28 days of age, were randomly allotted to 3 dietary groups of 5 replicate pens per group for 4 weeks of feeding. Experimental diets were: i) zinc deficient diet (ZnD, 24 mg/kg Zn supplementation from ZnSO4), ii) inorganic Zn diet supplemented with 120 mg/kg of Zn from Zn sulfate (ZnSO4), and iii) organic Zn diet supplemented with 120 mg/kg of Zn from PC-Zn. The body weight of pigs were recorded at the beginning, at the middle and at the end of the experiment, and the amount of feed supplied each day was recorded. Five barrows from each dietary treatment group were selected to be anesthetized and euthanized at the end of the trial to determine the Zn, Cu, Fe, and Mn concentrations, the hepatic metallothionein content, the levels of methane dicarboxylic aldehyde (MDA), Mn, and Cu/Zn superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in the spleen, the levels of interleukin (IL)-2, IL-4, IL-10, interferon (IFN)-γ, CD3+, CD4+, and CD8+ T lymphocyte. Results: The accumulation of Zn in the spleen, levels of SOD, GSH-Px, IL-4, IL-10, the proportions of CD3+ and CD4+ T lymphocyte, and the ratio of CD4+/CD8+ T lymphocyte were increased by organic Zn supplementation compared to ZnD, while the levels of MDA, IFN-γ, and proportion of CD8+ T lymphocyte were lowered. Conclusion: These findings indicate that Zn can improve the antioxidant potential and immune functions of weaned piglets.

Effects of Casein and Protein-free Diets on Endogenous Amino Acid Losses in Pigs

Zhang, Yongcheng,Li, Defa,Fan, Shijun,Piao, Xiangshu,Wang, Jitan,Han, In K. Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.11

Quantification of endogenous amino acid loss at the terminal ileum is an essential means for calculation of the true amino acid digestibility of a feedstuff. Since nitrogen appeared in the determined diet or not could shift the results very much, also, none of digestibility markers could be recovered with 100% rate at the terminal ileum, the objectives of the present study were: (1) to determine endogenous amino acid losses when fed either a casein diet or a protein-free diet and (2) to examine the reliability of chromic oxide or acid insoluble ash in the protein-free diet. Six ileal-cannulated pigs ($65{\pm}1.85 kg$ BW) with a simple T-cannula in the terminal ileum were used in a replicated $3{\times}3$ Latin square designed trial, after allowed a 14 d recuperation period. Each test period ran for 12 days comprised of a 10 d adjustment period and a 2 d collection period. The endogenous AA losses of His, Ile, Lys, Cys, Thr, Val, Trp, Asp, Glu, and Ser from pigs fed the casein diet were significantly higher than those of the protein-free diet (p<0.05). No significant difference was found in the amount of endogenous amino acid loss when determined with the different markers in the protein-free diet (p>0.05). These data suggest that endogenous amino acid loss could be underestimated when a protein-free diet is used. A direct effect of dietary peptides on the endogenous amino acid loss was found when the casein diet was fed. Our results also indicate that acid insoluble ash can be used as an inert marker as an alternative to chromic oxide when measuring endogenous amino acid loss.

Evaluation of available energy and total tract digestibility of acid-hydrolyzed ether extract of cottonseed oil for growing pigs by the difference and regression methods

Jinbiao Zhao,Zhongchao Li,Mingbin Lyu,Ling Liu,Xiangshu Piao,Defa Li 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.5

Objective: The objective of this study was to determine the effect of inclusion level on the digestible energy (DE), metabolizable energy (ME), and total tract digestibility of acid-hydrolyzed ether extract (AEE) of cottonseed oil when fed to growing pigs. Methods: Forty-two barrows (initial body weight = 35.51±2.01 kg) were randomly allotted to a completely randomized design with a corn-soybean meal basal diet, five levels of cottonseed oil (2%, 4%, 6%, 8%, and 10%) and a 10% soybean oil diet. Each diet was replicated six times with one pig per replicate. The experiment lasted 19 days, 7 d for cage adaptation, 7 d for diets adaptation and last 5 d for feces and urine collection. The energy values and apparent total tract digestibility (ATTD) of cottonseed oil and soybean oil were calculated by the difference method, and regression equations were established to predict the energy values of cottonseed oil. The apparent digested fat of the entire intestinal tract was also regressed against dietary fat intake to determine the true total tract digestibility (TTTD) and endogenous loss of fat for cottonseed oil. Results: The results showed that the DE and ME contents of cottonseed oil were not different as the inclusion level increased. The DE and ME values determined by the regression equation were 36.28 MJ/kg and 34.96 MJ/kg, respectively, and the values were similar to the mean DE and ME values calculated by the difference method (36.18 and 35.56 MJ/kg, respectively). The ATTD of cottonseed oil was also not affected by the inclusion level of cottonseed oil, and the TTTD and EFL determined by the regression method were 92.40% and 13.83 g/kg of dry matter intake for corn-soybean basal diet. The DE, ME, and ATTD of AEE in soybean oil determined by the difference method were 35.70 MJ/kg, 35.20 MJ/kg and 92.31%, respectively. There were no differences in the DE, ME, and ATTD between cottonseed oil and soybean oil, although the ratio of unsaturated to saturated fatty acids for soybean oil was higher than for cottonseed oil. Conclusion: The DE, ME, and ATTD values of cottonseed oil were not affected by its dietary inclusion level. The energy values of cottonseed oil determined by the difference and regression methods were similar. Furthermore, the ratio of unsaturated to saturated fatty acid for oils was not the decisive factor to influence the energy values and ATTD of oils.