• Title/Summary/Keyword: Protein supplementation

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Intestinal segment and vitamin D3 concentration affect gene expression levels of calcium and phosphorus transporters in broiler chickens

  • Jincheng Han;Lihua Wu;Xianliang Lv;Mengyuan Liu;Yan Zhang;Lei He;Junfang Hao;Li Xi;Hongxia Qu;Chuanxin Shi;Zhiqiang Li;Zhixiang Wang;Fei Tang;Yingying Qiao
    • Journal of Animal Science and Technology
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    • v.65 no.2
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    • pp.336-350
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    • 2023
  • Two experiments were conducted in this research. Experiment 1 investigated the spatial expression characteristics of calcium (Ca) and phosphorus (P) transporters in the duodenum, jejunum, and ileum of 21-day-old broilers provided with adequate nutrient feed. Experiment 2 evaluated the effects of dietary vitamin D3 (VD3) concentration (0, 125, 250, 500, 1,000, and 2,000 IU/kg) on growth performance, bone development, and gene expression levels of intestinal Ca and P transporters in 1-21-day-old broilers provided with the negative control diet without supplemental VD3. Results in experiment 1 showed that the mRNA levels of calcium-binding protein 28-kDa (CaBP-D28k), sodium-calcium exchanger 1 (NCX1), plasma membrane calcium ATPase 1b (PMCA1b), and IIb sodium-phosphate cotransporter (NaPi-IIb) were the highest in the broiler duodenum. By contrast, the mRNA levels of inorganic phosphate transporter 1 (PiT-1) and 2 (PiT-2) were the highest in the ileum. Results in experiment 2 showed that adding 125 IU/kg VD3 increased body weight gain (BWG), feed intake (FI), bone weight, and percentage and weight of Ca and P in the tibia and femur of 1-21-day-old broilers compared with the negative control diet (p < 0.05). The rise in dietary VD3 levels from 125 to 1,000 IU/kg further increased the BWG, FI, and weights of the bone, ash, Ca, and P (p < 0.05). No difference in growth rate and leg bone quality was noted in the broilers provided with 1,000 and 2,000 IU/kg VD3 (p > 0.05). Supplementation with 125-2,000 IU/kg VD3 increased the mRNA abundances of intestinal Ca and P transporters to varying degrees. The mRNA level of CaBP-D28k increased by 536, 1,161, and 28 folds in the duodenum, jejunum, and ileum, respectively, after adding 1,000 IU/kg VD3. The mRNA levels of other Ca and P transporters (PMCA1b, NCX1, NaPi-IIb, PiT-1, and PiT-2) increased by 0.57-1.74 folds by adding 1,000-2,000 IU/kg VD3. These data suggest that intestinal Ca and P transporters are mainly expressed in the duodenum of broilers. Moreover, the addition of VD3 stimulates the two mineral transporter transcription in broiler intestines.

The Effects of Supplemental Bacterial Phytase to the Calcium and Nonphosphorus Levels in Feed of Laying Hens (산란계 사료 내 칼슘 및 무기태 인 수준에 따른 Bacterial Phytase 급여 효과)

  • Kang, H.K.;Park, S.Y.;Yu, D.J.;Kim, J.H.;Kang, G.H.;Na, J.C.;Kim, D.W.;Suh, O.S.;Lee, S.J.;Lee, W.J.;Kim, S.H.
    • Korean Journal of Poultry Science
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    • v.35 no.2
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    • pp.143-151
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    • 2008
  • This study was conducted to identify the correlation of bacterial phytase ($Transphos^{(R)}$) to the calcium level in feed. Of all 21-week-old 720 HyLine brown laying hens, 2 birds of similar weight were placed on each individual cage. The experiment was conducted by $3{\times}2{\times}3$ factorial design with including 3 different levels of phytase (0, 300, and 1,000 DPU/kg), 2 different levels of calcium (3.5% and 4.0%), and 3 different levels of no NPP addition 0% (0.095 NPP), 0.5% (0.185% NPP), and 1.0% (0.275% NPP). The feeding trial maintained the ME level of 2,800 kcal/kg and 16% for crude protein. The diet was fed ad libitum and 17 hours of lighting was provided throughout the experimental period. Egg production seemed to increase, in the 300 DPU of bacterial phytase added group and the cracked egg tended to reduce in Transphos added group. The egg productivity between treatment groups did not show significant difference by dietary calcium level, whereas non NPP added group (0.095% NPP) was found to be low compared to NPP added groups (P<0.05). The highest mean egg weight and the highest daily egg mass were detected in 300 DPU phytase added group. Although the mean egg weight was significantly higher in treatment groups fed with 3.5% calcium containing feeds (P<0.05), daily egg mass was no among treatment groups. The mean egg weight and daily egg mass were the lowest in non NPP added group (0.095% NPP) compared to other treatment groups (P<0.05). The feed intake showed similar pattern regardless of the bacterial phytase and calcium levels in the diet. However, the treatment groups fed diets containing NPP level of 0.275% and 0.165% showed significantly higher feed intake than the group fed with 0.095% NPP (P<0.05). Although the feed conversion was not affected by calcium and NPP levels in the diet, the most improved result was obtained from 300 DPU phytase added group (P<0.05). The eggshell breaking strength and thickness increased as dietary calcium level increase the level of calcium increases in diet. The treatment groups fed diet containing 0.275% and 0.165% NPP revealed to show improvement in eggshell breaking strength and yolk color index compared to the NPP non added (0.095% NPP) treatment group. The result of the present study suggests that the appropriate level of microbial phytase is 300 DPU and at this level, tricalciumphosphate supplementation in feed can be reduced to 40% of NRC recommendation. Higher calcium level in feed fail to show synergistic effect by adding microbial phytase.

Effect of Byproducts Supplementation by Partically Replacing Soybean Meal to a Total Mixed Ration on Rumen Fermentation Characteristics In Vitro (대두박 대체 부산물 위주의 TMR 사료가 반추위 내 미생물의 In Vitro 발효특성에 미치는 영향)

  • Bae, Gui Seck;Kim, Eun Joong;Song, Tae Ho;Song, Tae Hwa;Park, Tae Il;Choi, Nag Jin;Kwon, Chan Ho;Chang, Moon Baek
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.2
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    • pp.129-140
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    • 2014
  • This study was performed to evaluate the effects of replacing basic total mixed ration (TMR) with fermented soybean curd, Artemisia princeps Pampanini cv. Sajabal, and spent coffee grounds by-product on rumen microbial fermentation in vitro. Soybean in the basic TMR diet (control) was replaced by the following 9 treatments (3 replicates): maximum amounts of soybean curd (SC); fermented SC (FSC); 3, 5, and 10% FSC + fermented A. princeps Pampanini cv. Sajabal (1:1, DM basis, FSCS); and 3, 5, 10% FSC + fermented coffee meal (1:1, DM basis, FSCC) of soybean. FSC, FSCS, and FSCC were fermented using Lactobacillus acidophilus ATCC 496, Lactobacillus fermentum ATCC 1493, Lactobacillus plantarum KCTC 1048, and Lactobacillus casei IFO 3533. Replacing dairy cow TMR with FSC treatment led to a pH value of 6 after 8 h of incubation-the lowest value measured (p<0.05), and FSCS and FSCC treatments were higher than SC and FSC treatment after 6 h (p<0.05). Gas production was higher in response to 3% FSC and FSCC treatments than the control after 4-10 h. Dry matter digestibility was increased 0-12 h after FSC treatment (p<0.05) and was the highest after 24 h of 10% FSCS treatment. $NH_3-N$ concentration was the lowest after 24 h of FSC treatment (p<0.05). Microbial protein content increased in response to treatments that had been fermented by the Lactobacillus spp. compared to control and SC treatments (p<0.05). The total concentration of volatile fatty acids (VFAs) was increased after 6-12 h of FSC treatment (p<0.05), while the highest acetate proportion was observed 24 h after 5% and 10% FSCS treatments. The FSC of propionate proportion was increased for 0-10 h compared with among treatments (p<0.05). The highest acetate in the propionate ration was observed after 12 h of SC treatment and the lowest with FSCS 3% treatment after 24 h. Methane ($CH_4$) emulsion was lower with A. princeps Pampanini cv. Sajabal and spent coffee grounds treatments than with the control, SC, and FSC treatments. These experiments were designed to replace the by-products of dairy cow TMR with SC, FSC, FSCS, and FSCC to improve TMR quality. Condensed tannins contained in FSCS and FSCC treatments, which reduced $CH_4$ emulsion in vitro, decreased rumen microbial fermentation during the early incubation time. Therefore, future experiments are required to develop a rumen continuous culture system and an in vivo test to optimize the percentages of FSC, FSCS, and FSCC in the TMR diet of the dairy cows.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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