CN113229410B - 一种促进幼龄反刍动物瘤胃发育的饲料添加剂及其饲粮 - Google Patents
一种促进幼龄反刍动物瘤胃发育的饲料添加剂及其饲粮 Download PDFInfo
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Abstract
本发明属于饲料添加剂领域,具体涉及一种促进幼龄反刍动物瘤胃发育的饲料添加剂及其饲粮。本发明首次提出了β‑羟基丁酸可作为促进幼龄反刍动物瘤胃发育的饲料添加剂,其中β‑羟基丁酸作为信号分子促进瘤胃上皮细胞对短链脂肪酸的转运吸收,刺激瘤胃上皮细胞生酮作用,增加瘤胃乳头宽度,提高幼龄反刍动物采食量和增重速度,改善饲料利用效率。本发明通过实验证实β‑羟基丁酸和烟酸具有协同作用,可用于缓解瘤胃上皮组织炎症反应、促进瘤胃上皮细胞生酮作用,促进短链脂肪酸生成。β‑羟基丁酸或其与烟酸联用可作为饲料添加剂,用在包括奶牛犊牛、肉牛犊牛、牦牛犊牛、绵羊羔羊、山羊羔羊等在内的幼龄反刍动物的饲养上。
Description
技术领域
本发明涉及饲料添加剂技术领域,具体地,涉及β-羟基丁酸及其与烟酸协同促进幼龄反刍动物瘤胃发育的应用。
背景技术
反刍动物的胃分为4部分,即瘤胃、网胃、瓣胃和皱胃。反刍动物幼龄阶段,瘤胃经历了由非反刍向反刍的生理功能转变。同时,饲粮从以液体饲粮为主,逐渐过渡到以固体饲粮为主;消化位置也由皱胃消化过渡到瘤胃消化;消化方式由化学消化过渡到微生物消化。瘤胃作为反刍动物特有的消化器官,瘤胃的发育程度直接影响到其成年后的采食和消化能力及生产性能的发挥。瘤胃上皮组织是其执行吸收、转运和代谢功能的重要部位,由瘤胃微生物产生的短链脂肪酸的85%由瘤胃上皮直接吸收并可为宿主提供60%-80%的所需代谢能。
现代工厂化、规模化、集约化的牛羊养殖生产中,新生犊牛羔羊由于胃肠道发育不成熟而具有极大的可塑性,也极易受到营养调控的影响而改变其后期产奶和育肥性能的发挥。当前,幼龄反刍动物的瘤胃健康机理研究相对滞后,并缺乏有效的调控手段,致使瘤胃潜能未得以充分发挥,严重制约了幼龄反刍动物的健康生长和高效生产。因此,调控瘤胃功能的尽早发育是保障幼龄动物健康高效生产的关键。
发明内容
本发明的目的是提供一种促进幼龄反刍动物瘤胃发育的饲料添加剂及其饲粮。本发明首次发现特定剂量的β-羟基丁酸在动物机体内以信号分子等形式参与到幼龄反刍动物瘤胃发育过程中,提高瘤胃短链脂肪酸含量及生长性能;并且发现与单独添加β-羟基丁酸相比,β-羟基丁酸与烟酸在缓解瘤胃上皮组织炎症反应、促进瘤胃上皮细胞生酮作用等方面具有协同作用,为促进幼龄反刍动物健康生长和高效生产奠定了基础。
本发明的目的是通过以下技术方案实现的:β-羟基丁酸单独作为饲料添加剂,及其与烟酸协同作为饲料添加剂,能促进瘤胃菌群对饲粮碳水化合物及蛋白质等营养物质的消化、促进瘤胃上皮组织对短链脂肪酸的吸收转运,并作为信号分子调控瘤胃上皮组织发育。
第一方面,本发明提供β-羟基丁酸在促进幼龄反刍动物的瘤胃上皮组织发育、提高幼龄反刍动物瘤胃内容物中普雷沃氏菌属和琥珀酸弧菌属相对丰度、提高瘤胃短链脂肪酸含量、增加幼龄反刍动物瘤胃乳头宽度、改善幼龄反刍动物生长性能中的应用。
在上述应用中,所述β-羟基丁酸作为饲料添加剂加入到幼龄反刍动物的饲粮中;所述β-羟基丁酸在饲粮中的添加量为0.2%-5%;
优选的,β-羟基丁酸在饲粮中的添加量为0.2%-1.0%;
或在上述应用中,所述β-羟基丁酸添加到饮水或液体饲粮中饲喂;所述β-羟基丁酸在饮水或液体饲粮中的饲喂量为1-10g/天/头;
优选的,β-羟基丁酸在饮水或液体饲粮中的饲喂量为3-6g/天/头。
第二方面,本发明提供β-羟基丁酸和烟酸在促进幼龄反刍动物瘤胃上皮组织发育、提高幼龄反刍动物瘤胃内容物中普雷沃氏菌属和琥珀酸弧菌属相对丰度、提高瘤胃短链脂肪酸含量及生长性能、增加幼龄反刍动物瘤胃乳头宽度中的应用。
第三方面,本发明还提供,β-羟基丁酸和烟酸在缓解瘤胃上皮组织炎症反应、促进瘤胃上皮细胞生酮作用中的应用。
在上述应用中,所述β-羟基丁酸和烟酸协同作为饲料添加剂添加到饲粮中混合均匀饲喂;所述β-羟基丁酸在饲粮中的添加量为0.2%-5%,所述烟酸在饲粮中的添加量为0.05%-0.5%;
优选的,所述β-羟基丁酸在饲粮中的添加量为0.2%-1.0%,所述烟酸在饲粮中的添加量为0.05-0.1%。
在上述应用中,所述β-羟基丁酸与烟酸添加到饮水或液体饲粮中饲喂;所述β-羟基丁酸在饮水或液体饲粮中的饲喂量为1-10g/天/头,所述烟酸在饮水或液体饲粮中的饲喂量为100-2000mg/天/头;
优选的,所述β-羟基丁酸在饮水或液体饲粮中的饲喂量为3-6g/天/头,所述烟酸在饮水或液体饲粮中的饲喂量为500-1000mg/天/头。
在上述应用中,所述幼龄反刍动物包括肉用犊牛、奶牛犊牛、牦牛犊牛、绵羊羔羊、山羊羔羊等。
第四方面,本发明提供一种促进幼龄反刍动物瘤胃发育的饲粮,β-羟基丁酸在饲粮中的添加量为0.2%-1.0%,烟酸在饲粮中的添加量为0.05-0.1%;
或者,在饮水或液体饲粮中,β-羟基丁酸的饲喂量为3-6g/天/头,烟酸的饲喂量为500-1000mg/天/头。
在本发明提供的饲粮中,所述饲粮还包含以下质量百分含量的组分:苜蓿草粉15%-25%、玉米40%-50%、豆粕18%-24%、麦麸3%-5%、脂肪粉1%-4%、磷酸氢钙0.3%-1.0%、石粉1.0%-1.5%、食盐0.3%-0.8%和预混料0.5%-1.5%。
本发明还提供一种包含β-羟基丁酸或β-羟基丁酸与烟酸的促进幼龄反刍动物瘤胃发育的液体饲粮,包含以下质量百分含量的组分:水80%-85%、全脂奶粉5%-8%、乳清粉4%-6%、脂肪粉3%-5%、柠檬酸钙0.3%-0.8%、食盐0.1%-0.3%和预混料0.3%-0.5%。
在上述饲粮或液体饲粮中,所述预混料为每千克饲粮含有:VA 30-50万IU、VD 3-5万IU、VE 1000-2000IU、VK 80-100mg、VB1 40-50mg、VB2 320-280mg、VB6 40-50mg、VB12 2-5mg、泛酸钙700-900mg、烟酸1800-2200mg、铁5-8g、铜0.5-1g、锰3-6g、锌6-10g、碘60-80mg、硒30-40g、钴50-70g。
根据本领域技术人员的理解,本发明还要求保护β-羟基丁酸及其与烟酸协同作作为羔羊饲料添加剂在提高羔羊采食量、提高羔羊增重速度或提升羔羊对饲料利用效率中的应用。
本发明的有益效果至少在于:
(1)本发明发现将β-羟基丁酸及其与烟酸协同使用作为促进瘤胃发育的饲料添加剂应用到幼龄反刍动物的饲粮中,能促进瘤胃乳头宽度的增加,有利于促进瘤胃形态学发育和扩大瘤胃内壁吸收面积,促进瘤胃上皮对短链脂肪酸的吸收效率;进而提高了羔羊采食量和增重速度,改善了饲料利用效率;
(2)本发明发现将β-羟基丁酸及其与烟酸协同使用作为促进瘤胃发育的饲料添加剂应用到幼龄反刍动物的饲粮中,提高瘤胃内容物中普雷沃氏菌属和琥珀酸弧菌属相对丰度,有利于瘤胃菌群对饲粮碳水化合物及蛋白质等营养物质的消化吸收,促进短链脂肪酸的产生,为瘤胃发育提供更多的供能底物;
(3)本发明发现将β-羟基丁酸及其与烟酸协同使用作为促进瘤胃发育的饲料添加剂应用到幼龄反刍动物的饲粮中,其中β-羟基丁酸能作为信号调控因子上调溶质载体家族16成员A3(SLC16A3)物质转运蛋白的mRNA表达水平,促进瘤胃上皮细胞对短链脂肪酸的转运;
(4)本发明发现将β-羟基丁酸及其与烟酸协同使用作为促进瘤胃发育的饲料添加剂应用到幼龄反刍动物的饲粮中,能作为信号调控因子上调生酮代谢限速酶HMGCS2及BDH1的表达水平,刺激瘤胃上皮细胞的生酮代谢,为瘤胃上皮细胞增殖供能;
(5)本发明发现将β-羟基丁酸与烟酸协同使用作为促进瘤胃发育的饲料添加剂应用到幼龄反刍动物的饲粮中,能作为信号调控因子上调瘤胃上皮细胞线粒体TRIM40基因的表达水平,缓解瘤胃上皮组织炎症反应。
附图说明
图1为本发明试验例1中不同剂量β-羟基丁酸及β-羟基丁酸与烟酸协同对羔羊瘤胃上皮组织基因mRNA表达的影响图。
具体实施方式
以下实例用于说明本发明,但不用来限制本发明的范围。在不背离本发明精神和实质的情况下,对本发明方法、步骤或条件所作的修改或替换,均属于本发明的保护范围。
若未特别指明,本发明实例中所用的实验材料、试剂、仪器等均可市售获得;若未具体指明,本发明实例中所有的技术手段均为本领域技术人员所熟知的常规手段。
实施例1
本实施例提供的饲养方法如下:羔羊在30-60日龄阶段不采食β-羟基丁酸,作为对照组。在试验过程中,根据羔羊的采食量和健康情况及时调整开食料饲喂量。同时,保证此阶段羔羊自由饮水。
实施例2
本实施例提供的饲养方法如下:羔羊在30-60日龄阶段每只羔羊每天采食β-羟基丁酸的量为3g。在试验过程中,根据羔羊的采食量和健康情况及时调整开食料饲喂量。同时,保证此阶段羔羊自由饮水。
实施例3
本实施例提供的饲养方法如下:羔羊在30-60日龄阶段每只羔羊每天采食β-羟基丁酸的量为6g。在试验过程中,根据羔羊的采食量和健康情况及时调整开食料饲喂量。同时,保证此阶段羔羊自由饮水。
实施例4
本实施例提供的饲养方法如下:羔羊在30-60日龄阶段每只羔羊每天采食β-羟基丁酸的量为9g。在试验过程中,根据羔羊的采食量和健康情况及时调整开食料饲喂量。同时,保证此阶段羔羊自由饮水。
实施例5
本实施例提供的饲养方法如下:羔羊在30-60日龄阶段每只羔羊每天采食β-羟基丁酸的量为6g,烟酸500mg。在试验过程中,根据羔羊的采食量和健康情况及时调整开食料饲喂量。同时,保证此阶段羔羊自由饮水。
试验例1
本试验例提供采用实施例1-5的方法饲养得到的羔羊的试验报告。
1、试验设计
选用出生日龄一致、体重相近的健康山羊羔羊72只,分成4组,每组18只,每组6个重复,每重复3只羔羊。并采用本发明实施例1-5的方法进行饲养试验。
在本发明的具体实施例中,所用开食料的组分如下:苜蓿草粉15%-25%、玉米40%-50%、豆粕18%-24%、麦麸3%-5%、脂肪粉1%-4%、磷酸氢钙0.3%-1.0%、石粉1.0%-1.5%、食盐0.3%-0.8%和预混料0.5%-1.5%。所述预混料为每千克饲粮含有:VA30-50万IU、VD 3-5万IU、VE 1000-2000IU、VK 80-100mg、VB1 40-50mg、VB2 320-280mg、VB640-50mg、VB12 2-5mg、泛酸钙700-900mg、烟酸1800-2200mg、铁5-8g、铜0.5-1g、锰3-6g、锌6-10g、碘60-80mg、硒30-40g、钴50-70g。
2、指标测定
(1)生长性能测定:准确称量羔羊30、60日龄体重,并记录每日开食料采食量,以计算平均日增重和采食量。
(2)瘤胃pH值及形态发育指标测定:60日龄时每处理组选取6只羔羊屠宰,分离羔羊瘤胃,用便携pH计测定瘤胃液pH值。使用图像采集系统和图像分析系统测量瘤胃乳头高度、乳头宽度等指标。
(3)瘤胃菌群多样性的指标:提取瘤胃内容物中DNA,扩增和纯化后构建Miseq文库并进行测序。采用RDP Classifier算法对OTU代表序列进行比对分析得到各种菌群物种信息。
(4)瘤胃上皮组织发育相关基因mRNA的表达:采用高通量测序技术检测瘤胃上皮组织差异表达基因,利用实时荧光定量PCR检测瘤胃上皮组织生酮关键酶(3-羟基丁酸脱氢酶1,BDH1)、线粒体羟甲基戊二酸单酰辅酶A合酶2,HMGCS2)、短链脂肪酸转运蛋白(溶质载体家族16成员A3,SLC16A3)及炎症蛋白(三重基序蛋白40,TRIM40)等的基因mRNA相对表达量。
3、试验结果
(1)羔羊生长性能
在60日龄时,实施例2、5组羔羊在体重和采食量方面均显著高于实施例1(对照组)(P<0.05);实施例2、3和5组羔羊在日增重方面显著高于实施例1(对照组)(P<0.05);实施例2-5组羔羊在料重比方面均显著低于实施例1(对照组)(P<0.05),表明β-羟基丁酸作为饲料添加剂提高了羔羊开食料的采食量和增重速度,并改善了饲料利用效率(表1)。
表1β-羟基丁酸对羔羊生长性能的影响
项目 | 实施例1(对照组) | 实施例2 | 实施例3 | 实施例4 | 实施例5 |
始重,kg | 5.21 | 5.54 | 5.18 | 5.03 | 5.32 |
末重,kg | 6.49<sup>a</sup> | 7.80<sup>b</sup> | 7.25<sup>ab</sup> | 6.75<sup>ab</sup> | 8.04<sup>b</sup> |
采食量(g/d) | 167.5<sup>a</sup> | 257.0<sup>b</sup> | 237.1<sup>ab</sup> | 213.6<sup>ab</sup> | 267.4<sup>b</sup> |
日增重(g/d) | 42.86<sup>a</sup> | 76.44<sup>b</sup> | 75.00<sup>b</sup> | 60.96<sup>a</sup> | 85.53<sup>c</sup> |
料重比 | 5.46<sup>b</sup> | 3.53<sup>a</sup> | 3.57<sup>a</sup> | 3.82<sup>a</sup> | 3.13<sup>a</sup> |
注:同行数据肩标不同小写字母代表显著性差异(P<0.05),下表同。
(2)羔羊瘤胃组织形态发育指标
通常瘤胃上皮乳头高度、乳头宽度等相关指标均可用来评定瘤胃的组织形态学发育。由表2可以看出,实施例2-5的方法饲养的羔羊的乳头宽度均显著大于实施例1(对照组)(P<0.05),分别提高了18.81%、19.58%、15.57%和48.26%;乳头宽度的增加扩大了幼龄反刍动物瘤胃内表面积,将有助于更好的吸收营养物质,并且β-羟基丁酸与烟酸联用呈现协同作用。
表2β-羟基丁酸对羔羊瘤胃组织形态发育指标的影响(单位:μm)
项目 | 实施例1(对照组) | 实施例2 | 实施例3 | 实施例4 | 实施例5 |
乳头高度 | 930.98 | 1147.30 | 1140.91 | 1251.79 | 1246.50 |
乳头宽度 | 275.63<sup>a</sup> | 327.48<sup>b</sup> | 329.6<sup>b</sup> | 315.79<sup>ab</sup> | 408.66<sup>c</sup> |
上皮厚度 | 78.22 | 73.52 | 68.45 | 85.04 | 77.05 |
角质层厚度 | 32.21 | 42.19 | 33.27 | 42.43 | 41.08 |
(3)羔羊瘤胃菌群组成
从表3可以看出,实施例3及5较实施例1(对照组)显著提高了羔羊瘤胃中普雷沃氏菌属(Prevotella)和琥珀酸弧菌属(Succinivibrio)的相对丰度(P<0.05)。普雷沃氏菌属和琥珀酸弧菌属可利用饲粮碳水化合物和蛋白质发酵产生丙酸及其他短链脂肪酸。二者相对丰度的提高,有利于瘤胃菌群对精料和粗饲料的分解利用及生成瘤胃短链脂肪酸,从而提高幼龄反刍动物对饲粮的消化吸收。
表3β-羟基丁酸对羔羊瘤胃菌群相对丰度的影响(单位:%)
项目 | 实施例1(对照组) | 实施例2 | 实施例3 | 实施例4 | 实施例5 |
普雷沃氏菌属 | 30.95<sup>a</sup> | 31.05<sup>a</sup> | 40.24<sup>b</sup> | 27.16<sup>a</sup> | 48.44<sup>b</sup> |
琥珀酸弧菌属 | 8.02<sup>a</sup> | 7.35<sup>a</sup> | 24.72<sup>b</sup> | 10.14<sup>a</sup> | 20.55<sup>b</sup> |
(4)羔羊瘤胃发酵参数
由表4可知,实施例3及5较其他实施例显著提高了(P<0.05)羔羊瘤胃TSCFA(总短链脂肪酸)、丙酸及丁酸含量,显著降低了(P<0.05)乙丙比(乙酸与丙酸比值)。瘤胃短链脂肪酸(总TSCFA、丙酸、丁酸)浓度的提高,可以给羔羊机体提供更多的能量底物,有利于瘤胃发育和生长性能的提升。
表4β-羟基丁酸对羔羊瘤胃发酵参数的影响
(5)羔羊瘤胃上皮组织发育相关基因表达
从附图1可以看出,β-羟基丁酸对羔羊瘤胃上皮组织发育相关基因表达有显著影响(P<0.05)。其中,HMGCS2基因表达量在实施例3、实施例5较实施例1均呈现出显著上调(P<0.05);BDH1及溶质载体家族16成员A3(SLC16A3)基因表达水平在实施例2,3,4,5组均呈现出显著上调(P<0.05);实施例5组羔羊的TRIM40的基因表达水平比实施例1均显著上调(P<0.05)。HMGCS2及BDH1基因表达的上调有利于瘤胃上皮细胞生酮作用和细胞增殖,SLC16A3的上调与瘤胃上皮细胞对短链脂肪酸的吸收有关,TRIM40的上调表达有利于缓解炎症反应。表明适宜剂量的β-羟基丁酸可作为信号因子通过调控瘤胃上皮细胞相关基因表达刺激瘤胃上皮细胞增殖,促进瘤胃整体发育,并且β-羟基丁酸与烟酸联用对缓解瘤胃上皮组织炎症反应,促进瘤胃上皮细胞线粒体生酮作用呈现协同作用。
综上所述,β-羟基丁酸作为促进幼龄反刍动物瘤胃发育的饲料添加剂,可:①调控瘤胃微生物区系优势菌群的丰度,有利于饲料营养物质的消化和短链脂肪酸的生成;②通过调控瘤胃上皮细胞转运蛋白及生酮代谢通路基因表达,刺激瘤胃上皮细胞增殖和组织发育;③通过增加瘤胃乳头宽度提高瘤胃内表面积和瘤胃整体发育,提高羔羊采食量及生长速度,并改善饲料利用效率。
β-羟基丁酸与烟酸协同作用,对于促进瘤胃形态发育、缓解瘤胃上皮组织炎症反应取得了更佳的作用。综合各项指标,以实施例3及实施例5提供的饲养方法对羔羊瘤胃上皮组织发育的促进效果较好。
对比例1
本对比例提供了使用丁酸作为羔羊饲料添加剂,与上述实施例3进行对比饲喂羔羊的试验结果。
选用30日龄、体重相近的健康羔羊36只,分成2组,每组18只,每组6个重复,每重复3只羔羊。羔羊分别采用6g/天/头的丁酸及本发明实施例3的方法进行饲养试验。所用试验饲粮与测定方法同试验例1。
结果表明,在60日龄时,实施例3的方法饲养的羔羊在日增重、菌群结构及瘤胃组织形态等指标方面均优于丁酸组(P<0.05)。本对比例的结果显示,与实施例3相比,丁酸虽然也具有促进羔羊采食量提升的效果,但没有对瘤胃微生物区系产生积极效果,甚至还降低了瘤胃pH,抑制了瘤胃上皮细胞TRIM40基因表达,意味着引起了瘤胃上皮组织的炎症反应。因此,外源添加丁酸作为饲料添加剂饲喂羔羊,对瘤胃菌群结构,尤其是瘤胃上皮组织健康未呈现明显的促进作用。
对比例2
本对比例提供了在不同的饲料组分下β-羟基丁酸对瘤胃发育的不同作用效果。
选用30日龄、体重相近的健康羔羊54只,分成3组,每组18只,每组6个重复,每重复3只羔羊。在试验例1所用试验饲粮的基础上,通过调整配方形成3个不同NDF水平的日粮。采用本发明实施例3的方法进行饲养试验。测定指标及方法参照试验例1。
所用饲粮的组分分别为:日粮1同试验例1;日粮2:苜蓿草粉0%-5%、玉米50%-65%、豆粕18%-24%、麦麸3%-5%、脂肪粉1%-4%、磷酸氢钙0.3%-1.0%、石粉1.0%-1.5%、食盐0.3%-0.8%和预混料0.5%-1.5%;日粮3:苜蓿草粉30%-40%、玉米30%-45%、豆粕18%-24%、麦麸3%-5%、脂肪粉1%-4%、磷酸氢钙0.3%-1.0%、石粉1.0%-1.5%、食盐0.3%-0.8%和预混料0.5%-1.5%。所用预混料同试验例1。
结果表明,在60日龄时,不同组分饲粮下,羔羊在日增重、菌群结构及瘤胃组织形态等指标方面均出现了显著变化,其中以本发明试验例1的日粮配方效果最好。本对比例的结果显示,β-羟基丁酸发挥作用与饲粮组分有密切关系。β-羟基丁酸作为饲料添加剂用于幼龄反刍动物的饲养中,需要搭配适宜的饲粮配方才能取得预期效果。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (5)
1.β-羟基丁酸和烟酸在制备提高幼龄反刍动物瘤胃内容物中普雷沃氏菌属和琥珀酸弧菌属相对丰度的饲粮中的应用;
所述β-羟基丁酸和烟酸协同作为饲料添加剂添加到饲粮中混合均匀饲喂;所述β-羟基丁酸在饲粮中的添加量为0.2%-5%,所述烟酸在饲粮中的添加量为0.05%-0.5%。
2.根据权利要求1所述的应用,其特征在于,
所述β-羟基丁酸在饲粮中的添加量为0.2%-1.0%,所述烟酸在饲粮中的添加量为0.05%-0.1%。
3.根据权利要求2所述的应用,其特征在于,所述β-羟基丁酸与烟酸添加到液体饲粮中饲喂;所述β-羟基丁酸在液体饲粮中的饲喂量为1-10 g/天/头,所述烟酸在液体饲粮中的饲喂量为100-2000 mg/天/头。
4.根据权利要求3所述的应用,其特征在于,所述β-羟基丁酸在液体饲粮中的饲喂量为3-6 g/天/头,所述烟酸在液体饲粮中的饲喂量为500-1000 mg/天/头。
5.根据权利要求1-4任一项所述的应用,其特征在于,所述幼龄反刍动物包括奶牛犊牛、牦牛犊牛、绵羊羔羊、山羊羔羊。
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