Greene Elizabeth, Mallmann Barbara, Wilson Jonathan W, Cowieson Aaron J, Dridi Sami
Department of Poultry Science, University of Arkansas, Fayetteville, AR, United States.
DSM Animal Nutrition and Health, Kaiseraugst, Switzerland.
Front Physiol. 2020 Jun 26;11:736. doi: 10.3389/fphys.2020.00736. eCollection 2020.
Phytate forms insoluble precipitates with various cations that are recalcitrant to digestion in poultry. Dietary supplementation with exogenous phytase has been shown to improve phytate solubility and digestibility and, in turn, improve animal growth performance. Although the kinetics of phytate hydrolysis by exogenous phytase are well described , the progression of the reaction is still not well defined. The aim of the present study was, therefore, to monitor the kinetic variation of myo-inositol (myo-Ins) levels in both circulation and feather following exogenous phytase supplementation. In experiment 1, 4 week-old male broilers were individually housed with access to water and a standard commercial diet. Birds were maintained under environmental temperature of 24°C and 30% RH. Birds were cannulated in the cutaneous ulnar vein on the right wing and remained untouched for 3 days. On the day of the experiment, birds were randomly divided into three body weight-matched groups and fed either the control diet, the control diet-supplemented with myo-Ins or Ronozyme HiPhos (0.06%, DSM Nutritional Products, Switzerland) for 10 h. In the experiment 2, birds were fed only HiPhos for 30 h. Growing feathers and blood were collected at baseline and then every 2 h for 10 h (experiment 1) and 30 h (experiment 2) post-prandially. Plasma and feather myo-Ins levels were determined by UHPLC-MS/MS. The relative expression of inositol polyphosphate-1-phosphatase (INPP1), inositol hexakisphosphate kinase 1-3 (IP6K1-3), inositol-3-phosphate synthase (ISYNA), and multiple inositol-polyphosphate phosphatase 1 (MNPP1) genes in blood and feathers was determined by real-time qPCR using 2 method. Plasma and feather myo-Ins levels were significantly increased by HiPhos at 6 h to 8 h post-prandial. The mRNA abundances of INPP1, IP6K1, and ISYNA in the circulation were significantly down regulated at all periods compared to the baseline levels. IP6K2, IP6K3, and MINPP1 gene expression, however, was up regulated at 8 h post-prandial and then returned to the baseline levels. In feathers, the expression of INPP1 was induced at 8 h post-prandial and remained higher compared to the baseline. The expression of IP6K2, IP6K3, and MINPP1 was down regulated during the first 10 h and then returned to baseline levels for the rest of the post-prandial period. Taken together, our data show that phytase modulates the expression of genes associated with myo-Ins metabolism and generates release of myo-Ins in both circulation and feather at 6-10 h post-feeding. Feather myo-Ins concentration could be used as a non-invasive method to monitor phytate hydrolysis in practice.
植酸盐会与多种阳离子形成不溶性沉淀,这些沉淀在家禽体内难以消化。研究表明,在日粮中添加外源植酸酶可提高植酸盐的溶解度和消化率,进而改善动物的生长性能。尽管外源植酸酶水解植酸盐的动力学已得到充分描述,但该反应的进程仍未完全明确。因此,本研究的目的是监测外源添加植酸酶后,循环系统和羽毛中肌醇(myo-Ins)水平的动力学变化。在实验1中,将4周龄的雄性肉鸡单独饲养,提供饮水和标准商业日粮。鸡饲养在环境温度为24°C、相对湿度为30%的环境中。在右翼的尺侧皮静脉进行插管,3天内不进行其他操作。在实验当天,将鸡随机分为三个体重匹配组,分别饲喂对照日粮、添加肌醇的对照日粮或Ronozyme HiPhos(0.06%,帝斯曼营养产品公司,瑞士)10小时。在实验2中,鸡只饲喂HiPhos 30小时。在基线时以及餐后10小时(实验1)和30小时(实验2)每2小时采集生长中的羽毛和血液。通过超高效液相色谱-串联质谱法(UHPLC-MS/MS)测定血浆和羽毛中的肌醇水平。使用2−ΔΔCT法通过实时定量PCR测定血液和羽毛中肌醇多磷酸-1-磷酸酶(INPP1)、肌醇六磷酸激酶1 - 3(IP6K1 - 3)、肌醇-3-磷酸合酶(ISYNA)和多肌醇多磷酸磷酸酶1(MNPP1)基因的相对表达。餐后6至8小时,HiPhos使血浆和羽毛中的肌醇水平显著升高。与基线水平相比,循环系统中INPP1、IP6K1和ISYNA的mRNA丰度在所有时间段均显著下调。然而,IP6K2、IP6K3和MINPP1基因表达在餐后8小时上调,然后恢复到基线水平。在羽毛中,INPP1的表达在餐后8小时诱导,并与基线相比保持较高水平。IP6K2、IP6K3和MINPP1的表达在最初10小时下调,然后在餐后剩余时间恢复到基线水平。综上所述,我们的数据表明,植酸酶在进食后6 - 10小时调节与肌醇代谢相关基因的表达,并在循环系统和羽毛中产生肌醇释放。羽毛中的肌醇浓度可作为一种非侵入性方法在实际中监测植酸盐的水解情况。