CN107205442A - 具有25‑羟基维生素d和抗氧化剂/抗炎剂的组合的伴侣动物饲料 - Google Patents
具有25‑羟基维生素d和抗氧化剂/抗炎剂的组合的伴侣动物饲料 Download PDFInfo
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- CN107205442A CN107205442A CN201680005768.5A CN201680005768A CN107205442A CN 107205442 A CN107205442 A CN 107205442A CN 201680005768 A CN201680005768 A CN 201680005768A CN 107205442 A CN107205442 A CN 107205442A
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Abstract
本发明涉及25‑羟基维生素D3(“25‑OH D3”)和抗氧化剂/抗炎剂(抗坏血酸、维生素E和角黄素)的组合用于制备能够改善在已经经受过度饲喂的伴侣动物中观察到的各种问题的预混物或饲料的用途。还提供了含有25‑OH D3和抗氧化剂/抗炎剂以及预混物的食物。
Description
发明简述
本发明涉及25-羟基维生素D(“25-OH D3”和/或“25-OH D2”)与抗氧化剂/抗炎剂(抗坏血酸、维生素E和类胡萝卜素)的组合用于伴侣动物(特别是猫和狗)食物。该营养补充剂的组合保护防止与摄食过量和相关的肥胖有关的各种不利影响。其还涉及用于伴侣动物的含有25-羟基维生素D和抗氧化剂/抗炎剂的组合的饲料和饲料预混物。
背景技术
摄食过量以及因此导致的肥胖可能发生在伴侣动物中,包括狗和猫。可能导致的过度饲喂诱导的疾病包括代谢紊乱,包括糖尿病和心血管问题。宠物主人并不总是控制提供给动物的食物量,有些动物会吃到超过饱食点。
在用作模型的家禽中,在摄食过量期间观察到升高的血浆葡萄糖浓度、非酯化脂肪酸浓度、极低密度脂蛋白浓度、三酰基甘油浓度、磷脂浓度、神经酰胺浓度和鞘磷脂浓度。此外,肝脏的和循环的神经酰胺及鞘磷脂积累,以及肝脏和脂肪组织中促炎IL-1β表达的上调系统性地显示了饱饲(feed-satiated)母鸡体内的脂毒性发展。神经酰胺是将某些营养物(即饱和脂肪)和炎性细胞因子(例如肿瘤坏死因子-α,TNFα)与细胞功能以及拮抗胰岛素信号传导的调控和线粒体功能关联的关键中间物。此外,由于神经酰胺对特别敏感细胞类型的毒性效应,神经酰胺具有破坏心脏、胰腺和脉管系统的能力。导致饱饲母鸡的受损卵巢机能障碍(包括卵泡闭锁、卵巢退化,以及循环雌二醇水平的下降)的脂毒性,更进一步被等级卵泡内的神经酰胺积累以及IL-1β、丝氨酸棕榈酰转移酶和鞘磷脂酶的转录本丰度的上调,但是抑制的蛋白激酶Akt激活而示例。因此,体内证据已经描绘了神经酰胺和IL-1β在肉种母鸡体内介导过度饲喂诱发的卵泡闭锁和卵巢退化进展中的作用。
(25-OH-D3的注册商标;可从DSM Nutritional Products,瑞士商购获得)已被用于促进伴侣动物的,特别是大型犬的骨骼和关节健康。
25-OH D3和角黄素的组合也已经被用于家禽。WO2010/057811(DSM IP ASSETS,BV)描述了用于改善家禽的孵化率、生育力并降低胚胎死亡率的此组合。该组合可以商标MAXICHICK商购获得。在该专利公布中既没有提到抗坏血酸和高维生素E水平的内含物,也没有提到用于改善对摄食过量相关的肥胖的不利影响的用途。
WO14/191153(DSM IP ASSETS B.V.)教导了角黄素,和维生素C、维生素E中的至少一种,硒,以及任选地百里酚、丁子香酚、香草醛和γ-萜品烯中的至少一种的组合可以改善免疫现状、骨骼健康、骨骼发育和生长以及饲料转化,特别是当禽群经受与疫苗接种相关的压力时。
在一些动物饮食中维生素C(抗坏血酸)通常不会作为补充剂被包括,因为许多动物可以在正常饲养条件下产生足够的维生素C。然而,维生素C已经在某些特定条件下使用,诸如在热应激情况下。
维生素E一般被添加到伴侣动物食物中。对于狗,建议的最小量以干物质为基础在36至50IU/kg的范围内。对于猫,最小量也以干物质为基础在38至50.7IU/kg的范围内。许多优质宠物食物具有高得多的维生素E含量,并且其可高达500IU/kg。
为了改善、预防或治疗受损的葡萄糖调节、胰岛素作用、心肌病,以及最终的腹水和猝死,需要减少或改善因伴侣动物的摄食过量导致的代谢和激素失调。
发明内容
已经发现,根据本发明,25-羟基维生素D(25-OH D3和/或25-OH D2)与抗氧化剂/抗炎剂的组合改善了当母鸡被自由采食饲喂时观察到的不良代谢状况。将母鸡用作自由采食饲喂的伴侣动物(诸如猫或狗)的模型,因为它们也会经历与摄食过量相关的肥胖。根据本发明还已经发现,传统上包含在食物中作为抗氧化剂的生物活性物也具有抗炎活性,所述抗炎活性减轻与摄食过量相关的肥胖症的不利影响。
还已经发现,25-OH D与抗氧化剂/抗炎剂的组合能够改善、降低与摄食过量相关的心血管问题的严重程度,或预防该等心血管问题。
由于25-OH D2和25-OH D3可以在施用后以类似方式作用,所以设想,它们中的任一个可以分别与抗氧化剂/抗炎剂组合使用,或者可以将25-OH D3和25-OH D2两者的混合物与抗氧化剂/抗炎剂组合使用。如果共同使用,25-OH D3:25-OH D2的比率不是本发明的关键部分。单独使用25-OH D3是优选的。
本发明的抗氧化剂/抗炎剂包括抗坏血酸、维生素E和类胡萝卜素的组合。因此,本发明的一个方面是25-OH D3、类胡萝卜素、维生素E和抗坏血酸的组合。另一个实施方案是25-OH D2、类胡萝卜素、维生素E和抗坏血酸的组合。另一个实施方案是25-OH D3、25-OH D2、类胡萝卜素、维生素E和抗坏血酸的组合。类胡萝卜素是选自由以下各项组成的组的至少一种:番茄红素、虾青素、隐黄质、β-胡萝卜素、叶黄素、玉米黄质和角黄素。最优选的是β-胡萝卜素。
本发明的另一方面是以下各项的组合:
a)25-OH D,
b)类胡萝卜素,所述类胡萝卜素选自由以下各项组成的组:番茄红素、虾青素、隐黄质、β-胡萝卜素、叶黄素、玉米黄质和角黄素,
c)维生素E,
d)抗坏血酸,以及
e)任选地还包含至少一种另外的选自由以下各项组成的组的生物活性成分:
维生素D、维生素B2、维生素B6、烟酸、锌、铜、锰和硒。优选地,25-OH D是25-OH D3。优选地,至少维生素D是另外的生物活性成分。有时,另外的生物活性成分至少包括维生素D和硒。在一些情况下,添加所有另外的生物活性成分。
本发明的另一方面是包含25-OH D2或25-OH D3或它们的混合物、抗坏血酸、维生素E和类胡萝卜素(优选地β-胡萝卜素)的组合的伴侣动物食物。
另一个实施方案是包含25-OH D、类胡萝卜素(优选地β-胡萝卜素)、维生素E和抗坏血酸的组合的伴侣动物食物,所述组合任选地还包含至少一种另外的选自由以下各项组成的组的生物活性成分:维生素D、维生素B2、维生素B6、烟酸、锌、铜、锰、硒,以及它们的组合。优选地,25-OH D是25-OH D3。有时,另外的生物活性成分至少包括维生素D和硒。在一些情况下,添加所有其他生物活性成分。
本发明的另一方面是用于伴侣动物食物的预混物,该预混物包含25-OH D、维生素E、抗坏血酸和类胡萝卜素(优选地β-胡萝卜素)的组合。优选地,25-OH D是25-OH D3。当饲喂给易于摄食过量或肥胖的动物时,预混物和随后的饲料改进/改善与摄食过量相关的不利代谢病症。在一些实施方案中,饲料和预混物还包含至少一种选自由以下各项组成的组的另外的生物活性成分:
维生素D、维生素B2、维生素B6、烟酸、锌、铜、锰、硒,以及它们的组合。有时,另外的生物活性成分至少包括维生素D和硒。在一些情况下,添加所有另外的生物活性成分。
本发明的另一方面是一种改善自由采食饲喂的伴侣动物增加的体重的量的方法,其包括:将25-OH D、类胡萝卜素(优选地β-胡萝卜素)、维生素C和维生素E的组合施用于所述动物。任选地,动物还可以饲喂上述组合和至少一种选自由以下各项组成的组的另外的生物活性成分:
维生素D、维生素B2、维生素B6、烟酸、锌、铜、锰、硒,以及它们的组合。有时,另外的生物活性成分至少包括维生素D和硒。在一些情况下,添加所有另外的生物活性成分。
当使用本发明的饲料时,动物可以自由采食饲喂,并且通常经历的不良效应将以较小程度经历或完全不经历。这消除了家庭中有多只宠物并且宠物主人试图管理这些动物中一些动物的体重的情况下遇到的问题。
在优选的实施方案中,将25-OH D、类胡萝卜素、维生素E和抗坏血酸的组合添加至含有完全营养的所有必要成分的基础饮食中,即该组合提供超生理量的组分。因此,这可以区别于提供用于仅满足营养需求,使得动物不是维生素或营养不足的一种或几种成分的先前的用途。
附图说明
图1示出响应与25-OH D3+抗氧化剂/抗炎剂组合的自由采食量(ad libitum feedintake)的肉种母鸡的葡萄糖清除率和胰岛素分泌。饲喂3周后,通过翅静脉向母鸡注射单剂量葡萄糖(0.5g/kg BW)。在输注葡萄糖后的指定时间点,通过翅静脉插管收集血样,其中n=3。
图2示出响应与25-OH D3和抗氧化剂/抗炎剂组合的自由采食量的肉种母鸡的组织白介素-1β含量和血浆IL-6水平。在饲喂试验10周后收集组织和血样。具有不同上标字母的均值显著不同(P<0.05),n=3。
图3示出响应与25-OH D3和抗氧化剂/抗炎剂内含物组合的自由采食量的肉种母鸡的组织STAT-3激活。在饲喂试验10周后收集组织和血样。具有不同上标字母的均值显著不同(P<0.05),n=3。
图4至图6示出了响应与25-OH D3和抗氧化剂/抗炎剂内含物组合的自由采食量的肉种母鸡尸体剖检期间拍摄到的照片。饲喂试验10周后,对母鸡进行尸体剖检。
图7示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的产蛋量的影响。
图8示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的存活率的影响。
图9示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的体重的影响。
图10是一系列照片,显示了在限制性采食量或自由采食量情况下饮食补充25-OH-D3+抗氧化剂/抗炎剂的死亡母鸡的心脏的总体形态。
图11是一系列照片,显示了在限制性采食量或自由采食量情况下饮食补充25-OH-D3+抗氧化剂/抗炎剂的死亡母鸡的心脏的总体形态。
图12是一系列心电图(EGC),其证明了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的EGC的影响。箭头指向图形中的不规律部分。
图13是一系列EGS,其显示了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的心律失常ECG图的影响。
图14示出照片和图表,证明了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的心肌纤维化的影响。(在35周龄时)
图15是示出饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的血浆IL-6和IL-1β浓度的影响的图。
图16是示出饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的心肌细胞凋亡的影响的照片(在35周龄时)。
如在本说明书和权利要求书通篇中使用的,以下定义适用:
“25-OH D”是指25-羟基维生素D的任何形式(即25-OH D2或25-OH D3,或它们的混合物)。25-OH D3特别指代25-羟基维生素D3;25-OH D2特别指代25-羟基维生素D2。
“维生素D”是指维生素D2、维生素D3或组合。单独使用维生素D3是优选的。
“伴侣动物”是指狗、猫、大鼠、兔子、观赏禽类、观赏鱼类和爬行动物。
“摄食过量”是过度饮食;动物不会自觉限制其进食。
“改善体重增加”是指当自由采食饲喂摄取本文所述的25-OH D3和抗氧化剂/抗炎剂的组合时,家禽将获取显著更低的体重的量。摄取该组合的动物可能比饲喂限制性饮食的动物增加更多的体重。
“抗坏血酸”和“维生素C”在整个说明书和权利要求书中可互换使用。
“基础饮食”意味着所使用的食物向动物供应足够的维生素和矿物质,使伴侣动物具有充分供应的维生素和矿物质。
“25-OH-D3+抗氧化剂/抗炎剂”是指25-OH D3、维生素E、类胡萝卜素和抗坏血酸的组合,该组合以在本说明书中所叙述的剂量范围在饲料中施用作为基础饮食添加剂。任选地,并且优选地,附加的生物活性成分选自由以下各项组成的组:维生素D、维生素B2、维生素B6、烟酸、泛酸、叶酸、生物素、锌、铜、锰、硒以及它们的组合添加到25-OH D3、维生素E、角黄素和抗坏血酸的组合中。
“猝死”是指个体禽在没有显示先兆迹象或疾病或创伤的情况下死亡。通过尸体剖检,该禽通常会具有无法识别的心血管问题。
1.代谢问题
与摄食过量相关的代谢问题,并且所述代谢问题可以通过使用本发明的饲料/预混物来减轻、减少或消除,包括:
a)减少体重增加和/或不再增重;
b)清除非酯化脂肪酸;
c)改善血浆血脂异常(甘油三酯、鞘磷脂和神经酰胺);
d)改善肝脏、腿部、胸肌和心脏中的甘油三酯和神经酰胺积累;
e)抑制组织促炎IL-1β产生和血浆IL-6浓度;
f)通过上调心脏中STAT-3(信号转导子和转录激活因子3)的磷酸化,来进行心脏保护和增强心脏功能;
g)抑制免疫细胞渗透到心脏内;
h)减少腹水发生率。
这些上述观察到的改善的状况导致降低的死亡率、改善的胰岛素信号传导、减少的脂毒性发展和系统性炎症,以及针对供能过载诱导的心脏发病机理的心脏保护机制的激活。
重要的是,根据本发明已经发现,25-OH D3、类胡萝卜素、维生素C和维生素E的组合可以减少当允许动物自由采食时增加的体重的量。因此,该组合可以被添加到伴侣动物的正常食物中,并且体重管理对于宠物主人来说可以更容易。
随着更少的体重增加,还观察到了与减少的肥胖有关的其他益处。
2.心血管问题
禽群中的禽会经历猝死,即死因不是轻易显现的。我们进一步研究了这一点,如在实施例4中所详述的。我们在具有或没有25-OH D3和抗氧化剂/抗炎剂的组合的情况下,在自由采食饲喂和限制性饮食饲喂的禽群中研究了这种现象。我们的一些结果如下。
已经发现,根据本发明,对于25-OH D3与抗氧化剂/抗炎剂的组合,已经饲喂25-OHD3与抗氧化剂/抗炎剂的自由采食饲喂的禽、猝死禽(即已经经历猝死的禽)具有较高的体重,但较低的相对肝脏、腹部脂肪和心脏重量。
在猝死禽中,自由采食饲喂引起心脏适应性肥大;并且一些肥大生长可能在病理上发展成心室扩张。因此,心脏需要更高的收缩力以维持泵送功能,从而满足递送氧到外周组织的血液供应需要。这种状况可能导致心力衰竭。
重要的是,我们发现25-OH-D3+抗氧化剂/抗炎剂减少了心脏病理进展,从而降低了自由采食饲喂的禽的心力衰竭发生率。因此,本发明的另一方面是25-OH-D3+抗氧化剂/抗炎剂用于减少导致猝死的心脏问题的量。
在限制性饲喂和自由采食饲喂的禽两者中,使用25-OH-D3+抗氧化剂/抗炎剂的禽表现出更少的适应性肥大生长,这支持以下假说:大多数过量养料可以被分配到肌肉,从而用于增加的泵送功能的心脏肥大生长无法满足更高生长速度(肌肉)的氧气供应需要,因此可能引起心律失常和心力衰竭。
看到25-OH-D3+抗氧化剂/抗炎剂具有以下作用:
●减少在死亡禽尸体剖检中观察到的心脏病发病率(扩张、心包积液、破裂)的发生率。
●减少ECG图的不规律发生量
●减少自由采食饲喂的肉种母鸡的心律失常
●改善心脏病发病引起的猝死
●改善自由采食饲喂的母鸡的心脏纤维化。
●改善自由采食饲喂的母鸡的慢性系统性炎症。
●改善具有限制性采食量或自由采食量的母鸡的心脏细胞凋亡。
由于这些症状被预期在伴侣动物中涉及与在被观测的禽中相同的途径,所以可以得出结论,相同的病理也发生在肥胖伴侣动物中。因此,25OH D3+抗氧化剂也可以保护伴侣动物的心血管系统。
3.剂量
在本发明的一个方面,将25-OH D3与抗氧化剂/抗炎剂的组合施用于维生素充足而不是维生素缺乏的动物。维生素充足状态优选地是由于使用为猫或狗提供至少最少量的维生素和矿物质的基础饲料。因此,除了基础饮食之外,优选地使用本发明的组合。
下面给出的较低剂量适用于已经达到理想体重的伴侣动物,因为其将有助于伴侣动物避免增加更多的不希望的体重。中等剂量适用于超重或趋于肥胖的动物。高剂量适用于那些已经变得肥胖的动物。所有这些剂量可以与减少的卡路里伴侣动物食物组合施用。
25-OH D3:25-OH D3的量可以在15至200μg/kg食物的范围内。优选地,25-OH D3的量为35至150μg/kg食物。对于含有低剂量的本发明组合的食物,35μg/kg 25-OH D3食物是优选的;对于含有中等剂量的该组合的食物,69μg/kg食物是优选的;并且对于含有高剂量的食物,150μg饲料是优选的。
维生素E:维生素E的量可以在40至400mg/kg食物的范围内。优选地,该量为80至300mg/kg食物。对于含有低剂量的本发明组合的食物,80mg/kg维生素E是优选的。对于具有中等剂量的食物,150mg维生素E是优选的;对于具有高剂量的食物,300mg/kg维生素E是优选的。
β胡萝卜素:角黄素的量可以在1至15mg/kg饲料的范围内。优选地,该量为3至12mg/kg饲料。对于含有低剂量的本发明组合的饲料,3mg/kg角黄素是优选的。对于具有中等剂量的饲料,6mg角黄素是优选的;对于具有高剂量的饲料,12mg/kg角黄素是优选的。
抗坏血酸:抗坏血酸的量可以在40至400mg/kg饲料的范围内。优选地,该量为100至300mg饲料。对于含有低剂量的本发明组合的饲料,100mg/kg抗坏血酸饲料是优选的。对于具有中等剂量的饲料,150mg抗坏血酸是优选的;对于具有高剂量的食物,300mg/kg抗坏血酸是优选的。
因此,本发明的特定优选的饲料包含以下剂量(所有量按照每kg饲料计):
优选的饲料#1:
25-OH D3:15-200μg,
维生素E:40-400mg,
β胡萝卜素:1-15mg;以及
抗坏血酸:40-400mg。
优选的饲料#2:
25-OH D3:35-150μg,
维生素E:80-300mg,
β胡萝卜素:3-12mg,以及
抗坏血酸:80-300mg
优选的饲料#3:(低剂量饲料)该饲料优选用于理想体重的动物,并且用于帮助维持体重。
25-OH D3:35μg
维生素E:80mg
β胡萝卜素:3mg
抗坏血酸:80mg。
优选的饲料#4(中等剂量饲料)该饲料对于中度至重度肥胖的伴侣动物是优选的:
25-OH D3:69μg
维生素E:150mg
β胡萝卜素:6mg
抗坏血酸:150mg。
优选的饲料#5(高剂量饲料)这种饲料对于严重肥胖的伴侣动物是优选的:
25-OH D3:150μg
维生素E:300mg
β胡萝卜素:12mg
抗坏血酸:300mg。
上述抗氧化剂/抗炎剂相对于彼此的比率可以在以下范围内:
·维生素E比维生素C的比率可以在1-10:10-1的范围内;优选地1-5:5-1;更优选地1.5:1至1:1.5。以及最优选地为1:1。
·维生素E或维生素C比β-胡萝卜素的比率可以在40:1至1:1的范围内;优选地20:1至1:1;以及更优选地为10:1至1:1。
优选的比率包括以下各项
维生素E | 维生素C | β胡萝卜素 |
40 | 40 | 1 |
20 | 20 | 1 |
10 | 10 | 1 |
任选的附加成分:
对于上面列出的每种饲料,可以添加至少一种附加成分。优选地添加至少一种,更优选地多于一种的以下成分。在其他实施方案中,添加所有以下成分:
维生素D3——通常其以约2500IU/kg饲料存在于家禽饮食中。根据本发明,如果需要,将维生素D的量增加至至少3000IU/kg。
维生素B2:其可以3至25mg/kg;优选地6至20mg/kg添加。对于低剂量饲料,6mg/kg是优选的。对于中等剂量饲料,14mg/kg是优选的;并且对于高剂量饲料,20mg/kg是优选的。
烟酸:其可以25至300mg/kg饲料添加。优选地,其在60至200mg/kg的范围内。对于低剂量饲料,60mg/kg是优选的。对于中等剂量饲料,120mg/kg是优选的;并且对于高剂量饲料,200mg/kg是优选的。
泛酸:其可以10至120mg/kg饲料添加。优选地,其在15至80mg/kg的范围内。对于低剂量饲料,15mg/kg是优选的。对于中等剂量饲料,30mg/kg是优选的;并且对于高剂量饲料,80mg/kg是优选的。
叶酸:其可以1至8mg/kg饲料添加。优选地,其在2至6mg/kg的范围内。对于低剂量饲料,2mg/kg是优选的。对于中等剂量饲料,4mg/kg是优选的;并且对于高剂量饲料,6mg/kg是优选的。
生物素:其可以0.05至1.0mg/kg饲料添加。优选地,其在0.2至0.8mg/kg的范围内。对于低剂量饲料,0.2mg/kg是优选的。对于中等剂量饲料,0.4mg/kg是优选的;并且对于高剂量饲料,0.8mg/kg是优选的。
锌:其可以50至300mg/kg饲料添加。优选地,其在70至250mg/kg的范围内。对于低剂量饲料,70mg/kg是优选的。对于中等剂量饲料,125mg/kg是优选的;并且对于高剂量饲料,250mg/kg是优选的。
铜:其可以5至50mg/kg饲料添加。优选地,其在10至30mg/kg的范围内。对于低剂量饲料,10mg/kg是优选的。对于中等剂量饲料,20mg/kg是优选的;并且对于高剂量饲料,30mg/kg是优选的。
锰:其可以50至300mg/kg饲料添加。优选地,其在80至270mg/kg的范围内。对于低剂量饲料,80mg/kg是优选的。对于中等剂量饲料,150mg/kg是优选的;并且对于高剂量饲料,270mg/kg是优选的。
硒:其可以0.05至0.6mg/kg饲料添加。优选地,其在0.1至0.4mg/kg的范围内。对于低剂量饲料,0.1mg/kg是优选的。对于中等剂量饲料,0.3mg/kg是优选的;并且对于高剂量饲料,0.5mg/kg是优选的。
可以制备预混合物以提供上述剂量和优选的剂量。形成本发明一部分的一种预混物被配制成使得1克预混合物被添加到一千克食物中,并且得到的食物含有任何上述给定剂量中所述的剂量。当然,各种成分的量可以变化为使得一千克预混料被添加到一公吨饲料中,并且所得食物含有任何上述给定剂量中所述的剂量。下面的实施例中有具体的说明。
此外,设想了本文规定的组合可以添加到任何市售的狗或猫食物中,因此存在的25-OH D3和抗氧化剂/抗炎剂的总量可以等于食物中原来存在的量加上如本文所规定的添加。也设想了如本文所规定的25-OH D3和抗氧化剂/抗炎剂是基础饮食的唯一添加剂,该基础饮食含有用于猫或狗营养的至少最低所需营养物。
提出以下非限制性实施例以更好地说明本发明。
实施例
实施例1
从商用鸡群获得总共三十只45周龄的肉种母鸡(ROSS 308)以进行研究。如表1所示配制基础肉种鸡日粮。所计算的营养组成如表2中所示。
表1:基础肉种鸡日粮的成分组成。
1所提供的矿物预混物(按照每kg用于治疗组1、2和3的饮食计):
Cu 18mg;I 1.1mg;Fe 80mg;Mn 150mg;Zn 125mg;以及Se 0.25mg。
2关于进一步细节请参考表2。
表2.维生素预混组合物(每千克饮食所提供的)
表3.基础肉种鸡日粮的计算的营养组成(%)。
组成 | %w/w |
粗蛋白 | 16 |
粗脂肪 | 4.2 |
钙 | 3.1 |
钠 | 0.16 |
总磷 | 0.64 |
总ME | 2910kcal/kg |
饮食补充有69mcg/kg的与抗氧化剂/抗炎剂(维生素E、抗坏血酸、角黄素)和富集水平(enriched level)的选定的维生素组合的25-OH D3,或不补充。根据如下饲喂方案(限制性和自由采食)将母鸡随机分配成3个治疗组:
基础饮食——限制性饲喂(140克/天)
基础饮食——自由采食饲喂
基础饮食——自由采食饲喂+69mcg/kg饮食25-OH-D3+抗氧化剂/抗炎剂
将它们分别容纳在放置于受控室中的钢丝笼中,该受控室具有14小时:10小时的光照:黑暗期和25±3℃的温度。水是可自由采食的。实验期持续10周。饲喂试验三周后,将一些禽用于相关血浆参数分析。在实验结束时,将母鸡安乐死并处死以进行组织样品收集以供进一步研究。
组织形态学的尸体剖检
测定脂质和鞘脂分布——血清和组织
测定组织促炎细胞因子
测定胰岛素抗性
收集胫骨以用于骨强度分析
收获心脏(心肌病)和骨骼肌(胸肌和腿肌)来进行肌病分析。
实施例2
结果与讨论
25-OH D3和抗氧化剂/抗炎剂抑制过度饲喂的肉种母鸡的肥胖和腹部脂肪
肉种母鸡能够在肝脏、脂肪组织和发育中的卵母细胞的卵黄中储存大量过剩能量(以甘油三酯的形式)。脂肪生成(即,葡萄糖转化为甘油三酯)主要发生在禽的肝脏中,并且涉及一系列连锁的酶催化反应,包括糖酵解、柠檬酸循环和脂肪酸合成。肝脂肪生成受营养和激素两者的控制,并且对饮食变化具有高响应性。脂肪组织主要用作具有很少脂肪生成活性的脂质的储存部位。禽体内肝脏对照脂肪组织的差异脂肪生成能力是关键转录因子甾醇调节元件结合蛋白-1(SREBP-1)的表达的函数。SREBP-1的基因在肝脏中高度表达,而在脂肪组织中表达至较低的程度。此外,诸如脂肪酸合成酶、苹果酸酶、乙酰辅酶A羧化酶、ATP柠檬酸裂解酶、甾醇酰辅酶A去饱和酶1等多种产脂酶的基因的表达直接受SREBP-1的影响。
自由采食饲喂的肉种母鸡比限制性饲喂的那些肉种母鸡增加更多的腹部脂肪。饮食补充25-OH D3和抗氧化剂/抗炎剂改善了自由采食饲喂对体重和组织重量的有害影响,特别是对相对脂肪组织重量(肥胖)的有害影响(表4)。25-OH D3和抗氧化剂/抗炎剂增强了自由采食饲喂的母鸡的胫骨强度。
表4. 25-OH D3和抗氧化剂/抗炎剂对自由采食饲喂的肉种母鸡的体重、肝重、腹部脂肪重量和胫骨强度的影响
25-OH D3和抗氧化剂/抗炎剂降低了死亡率并改善了产蛋量、卵巢形态和血浆17β雌二醇水平
雌二醇的分泌是成功排卵卵泡的标志。除了触发排卵前期促性腺激素峰的作用外,雌激素还是重要的卵巢内生长、分化和生存因子。包含25-OH D3和抗氧化剂/抗炎剂降低了自由采食的禽中的卵巢变性和卵巢肿瘤样形态,增加了产蛋量并且保持了血浆雌二醇水平。
25-OH D3和抗氧化剂/抗炎剂改善了受损的葡萄糖清除率和胰岛素敏感性
饮食包含25-OH D3和抗氧化剂/抗炎剂可以改善胰岛素抗性,如通过改善过度饲喂10周的母鸡的空腹血浆葡萄糖和非酯化脂肪酸水平所证明(表6)。在葡萄糖清除率试验中,瘦母鸡在葡萄糖输注后30至60分钟之间表现出非常急剧的清除率,相反,肥胖母鸡在30至90分钟之间具有极缓慢的清除率(图1)。在胰岛素分泌方面,与瘦母鸡相比,肥胖母鸡在空腹状态和葡萄糖输注后表现出较高的血浆胰岛素水平(图1)。葡萄糖清除率和葡萄糖诱导的胰岛素分泌两者均通过在过度饲喂3周的母鸡体内引入25-OH D3和抗氧化剂/抗炎剂校正(图1)。
表5. 25-OH D3和抗氧化剂/抗炎剂对自由采食饲喂的肉种母鸡的血浆葡萄糖、非酯化脂肪酸(NEFA)和胰岛素的影响
25-OH D3和抗氧化剂/抗炎剂改善血脂异常
自由采食饲喂的母鸡具有升高的血浆甘油三酯、神经酰胺和鞘磷脂水平。然而,补充组合的25-OH D3和抗氧化剂/抗炎剂降低了自由采食饲喂母鸡血浆中这些脂质代谢物的水平(表6)。
表6. 25-OH D3和抗氧化剂/抗炎剂对自由采食饲喂的肉种母鸡的血浆三酰基甘油、神经酰胺和鞘磷脂的影响
25-OH D3和抗氧化剂/抗炎剂减少了组织甘油三酯和神经酰胺含量的积累
相较于自由采食饲喂的那些母鸡,饲喂补充的25-OH D3和抗氧化剂/抗炎剂的母鸡的肝脏、心脏和腿部肌肉中的甘油三酯和神经酰胺积累更低(表7)。
表7 25-OH D3和抗氧化剂/抗炎剂对自由采食饲喂的肉种母鸡的组织三酰基甘油和神经酰胺含量的影响
25-OH D3和抗氧化剂/抗炎剂抑制过度饲喂的肉种母鸡的组织促炎IL-1β产生和血浆IL-6浓度
肥胖相关的炎症被饮食25-OH D3和抗氧化剂/抗炎剂补充剂改善,如通过抑制脂肪组织、肝脏、腿部肌肉和胸肌以及心脏中的循环IL-6水平和IL-1β产生所证明(图2)。
25-OH D3和抗氧化剂/抗炎剂改善自由采食饲喂的肉肉种母鸡的脂肪毒性
肥胖的中心并发症是胰岛素抗性的发展,胰岛素抗性的发展是当胰岛素不能在其主要靶组织、骨骼肌和肝脏中引起餐后营养物储存时发生的。不希望受理论束缚,看似两种可能的机制可以解释增加的脂肪储存如何影响整个身体的整体胰岛素敏感性,从而有助于下调外周组织中的胰岛素信号传导。第一,递送到细胞或组织中的营养物超过了其储存容量,从而这会导致抑制胰岛素作用的代谢物的产生。特别重要的是,脂质衍生物,诸如三酰基甘油和神经酰胺,已被证明抑制特异性胰岛素信号传导中间体,从而阻断餐后葡萄糖摄取和/或糖原合成。在肉种雌鸡自由采食饲喂的情况下,这些代谢物在外周组织中的持续积累可以有助于整个母鸡体内的胰岛素抗性的持续状态和脂毒性发展的持续状态。第二,增加的肥胖诱导慢性炎症状态,慢性炎症状态的特征在于由脂肪细胞产生的或由浸润脂肪垫的巨噬细胞产生的促炎细胞因子循环水平的升高。这些炎症介质已经表现为直接拮抗胰岛素信号传导,并且还诱导分解代谢过程,从而进一步增加营养代谢物向胰岛素反应性器官的递送。
总体而言,导致在非脂肪组织中形成过量饱和脂肪酸以及因此脂质积累的葡萄糖过量供应提高了活性脂质(鞘脂)的细胞水平,活性脂质(鞘脂)抑制代谢调节所涉及的信号传导通路以及激活的炎症反应和脂毒性发展。具体地,神经酰胺是将过量营养物(即,饱和脂肪酸)和炎性细胞因子与胰岛素抗性的诱导相关联的推定中间体。此外,神经酰胺在各种不同细胞类型中是有毒的,并且能够损害心脏、胰腺和脉管系统。此外,25-羟基D3和抗氧化剂/抗炎剂有效改善由于在饱饲的肉种母鸡中发生的增加肥胖而引起的代谢和内分泌失调以及促炎性反应的有害效应。
25-OH D3和抗氧化剂/抗炎剂改善过度饲喂的肉种母鸡的心脏病发病、腹水和炎症
由于过量脂质积累,心脏可能会变得具有功能障碍。自由采食饲喂促进心脏中甘油三酯的积累,表明增加的心脏脂肪酸可用性被适应性地酯化成甘油三酯。此外,由于自由采食饲喂,心脏的神经酰胺含量也增加。神经酰胺是脂毒性心肌病中的心脏毒素,其引起炎症反应,如通过免疫细胞的更多心脏浸润所证明的。(表9)
表8. 25-OH D3和抗氧化剂/抗炎剂对自由采食饲喂的肉种母鸡的心脏反应的影响
心脏肥大在临床上代表对增加的心脏负担的适应性反应。然而,心脏对神经和激素因子的反应也可以独立于后负荷或血管阻力的增加而引起肥大变化。养料(Fuel)过载诱导的心脏补偿生长发生在肉种母鸡体内(表8)。心脏肥大可能变得适应不良,并且最终发展成病理性病症,从而导致心力衰竭。这些结果支持以下事实:即心脏中的脂毒性发展和肥厚生长倾向于引起炎症反应。
磷酸化的STAT3(信号转导子和转录激活因子3)的心脏保护作用近年来变得越来越明显。有趣的是,组合的25-OH D3和抗氧化剂/抗炎剂比限制性饲喂的肉种母鸡诱导更大的在心脏中的STAT-3激活(即STAT-3的磷酸化)(图3),其中在自由采食饲喂的肉种母鸡中观察到了最低的激活。当补充组合的25-羟基D3和抗氧化剂/抗炎剂时,自由采食饲喂的肉种母鸡体内的心包膜渗出液、心室扩张和腹水的发生率得到缓解。
实施例3
预混物
下表9呈现将要添加到饲料中的本发明组合物的一些最终剂量范围。
范围 | 低 | 中 | 高 | ||
维生素 | 单位 | 每kg饲料 | 每kg饲料 | 每kg饲料 | 每kg饲料 |
25-OH-D3 | mcg | 15-200 | 35 | 69 | 150 |
维生素E | mg | 40-400 | 80 | 150 | 300 |
角黄素 | mg | 1-15 | 3 | 6 | 12 |
维生素C | mg | 40-400 | 100 | 150 | 300 |
下表10提供了每1kg饲料的预混物的量:
表11:1克预混物可以用以下成分制成:
结论
补充的25-OH D3和抗氧化剂/抗炎剂通过以下方式来改善与肉种雌鸡过度饲喂有关的有害效应:
降低死亡率并改善卵巢功能,从而降低过度饲喂的肉种母鸡的繁殖性能。
改善内分泌(胰岛素)信号传导
减少脂毒性发展和系统性炎症
激活抗供能物过载诱导的心脏发病机理的心脏保护机制
实施例4
食物
干燥的宠物食品配方
向该配方中添加25-OH D3、β胡萝卜素、维生素C和维生素E。润湿的宠物食品配方
向该配方中添加25-OH D3、β胡萝卜素、维生素C和维生素E。
实施例4
心肌病试验
实施例5
心肌病试验
材料和方法
从商用鸡群获得总共三十只45周龄的肉种母鸡(ROSS 308)以进行研究。如表12所示配制基础肉种鸡日粮。所计算的营养组成如表13中所示。
表12:基础肉种鸡日粮的成分组成。
1所提供的矿物预混物(按照用于治疗组1、2和3的每kg饮食计):Cu,18mg;
I,1.1mg;Fe,80mg;Mn,150mg;Zn,125mg;以及Se,0.25mg。
2参考表13,下文为进一步细节。
表13.维生素预混组成(每千克饮食所提供的)
表14.经计算的基础肉种鸡日粮的营养组成(%)。
饮食以69meg/kg饮食的25-OH-D3补充与抗氧化剂(抗坏血酸、角黄素)和富集水平的选定的维生素组合的或不补充。根据如下饲喂方案(限制性和自由采食)将母鸡随机分配成多个治疗组:
1.基础饮食——限制性饲喂(140克/天)
2.基础饮食——限制性饲喂+(69mcg/kg饮食25-OH-D3)+抗氧化剂/抗炎剂
3.基础饮食——自由采食饲喂
4.基础饮食——自由采食饲喂+(69mcg/kg饮食25-OH-D3)+抗氧化剂/抗炎剂
结果:
表15.饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的产蛋量的影响。
PROD=产蛋率(鸡蛋/天/母鸡,%);YIELD=产蛋量(鸡蛋/母鸡)
结果以均值±SEM表示。
相同鸡群内具有不同上标字母的均值有明显差异(P<0.05)。
图7示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的产蛋量的影响。从表15和图7可以得出结论:
1.25-OH-D3+抗氧化剂/抗炎剂通过促进存活来提高总产蛋量。
2.在死亡禽中,25-OH-D3+抗氧化剂/抗炎剂对自由采食饲喂的禽的产蛋量和产蛋率没有影响,但减少了限制性饲喂的禽的产蛋量和产蛋率。然而,在整个禽群中,25-OH-D3+抗氧化剂/抗炎剂提高了产蛋量,而不提高产蛋率。
3.这些结果表明,25-OH-D3+抗氧化剂/抗炎剂加速了易猝死的限制性饲喂禽的死亡进程,从而作为禽群挑选工具来排除易猝死禽以获得更长的存活时间,并从而降低禽群养护成本。
表16.饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的死亡率和死亡母鸡的身体特征的影响
结果以均值±SEM表示。
具有不同上标字母的均值有显著差异(P<0.05)。
图8示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的存活率的影响。
从表6和图8可以得出结论:
1. 25-OH-D3+抗氧化剂/抗炎剂提高了禽存活率。
2.在自由采食饲喂的禽中,使用25-OH-D3+抗氧化剂/抗炎剂的死亡禽具有较高的体重,但较低的相对肝脏、腹部脂肪和心脏重量,这表明大多数过量养料可以被分配到肌肉,从而用于增加泵送功能的心脏适应性肥大生长无法满足更高生长速度(肌肉)的氧气供应需要,因此可能引起心律失常和心力衰竭。
表17.饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的35周龄和50周龄肉种母鸡的屠体性状的影响
结果以均值±SEM表示。
具有不同上标字母的均值差异显著(P<0.05)。
*:与35周龄具有显著差异。
自由采食饲喂的50周龄禽中的低的相对肝重似乎是由于卵巢退化发展以及由此导致的雌激素分泌减少,从而导致肝脏中用于卵黄沉积的脂质合成减少。
图9示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的体重的影响。
表18.饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的死亡母鸡的心脏病发病率的影响
结果表示为比率。
图10至图11示出了在限制性采食量或自由采食量情况下饮食补充25-OH-D3+抗氧化剂/抗炎剂的死母鸡的心脏的总体形态。
从表17和表18以及图9至图11得出的结论和注释:
1.自由采食饲喂引起心脏适应性肥大;并且一些肥大生长可能在病理上发展成心室扩张。因此,心脏需要更高的收缩力以维持泵送功能,从而满足递送氧到外周组织的血液供应需要,从而可能引发心力衰竭。
2. 25-OH-D3+抗氧化剂/抗炎剂降低死亡禽中的心脏病发病率(扩张、心包积液、破裂)的发生率。
3.在限制性和自由采食饲喂的禽中,使用25-OH-D3+抗氧化剂/抗炎剂的禽表现出较少的适应性肥大生长。这支持以下假说:大多数过量养料可以被分配到肌肉,从而用于增加的泵送功能的心脏的肥大生长无法满足更高生长速度(肌肉)的氧气供应需要,因此可能引起心律失常和心力衰竭。
表19.饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的心电图(ECG)图形和心律失常的影响。关于EGC图的示例,请参见图12和图13。
从表19以及图12至图13得出的结论和注释:
25-OH-D3+抗氧化剂/抗炎剂减少自由采食饲喂的肉种母鸡的ECG图(图D至图G)的不规律发生和心律失常,并改善心脏病发病诱发的猝死。
图14示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的心肌纤维化的影响。(在35周龄时)条形上方标有字母的均值有显著差异(P<0.05)。
从图14得出的结论和注释:
25-OH-D3+抗氧化剂/抗炎剂改善自由采食饲喂的母鸡的心肌纤维化。图15示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的血浆IL-6(上部图)和IL-1β浓度(下部图)的影响。结果以均值±SEM(n=6)表示。条形上方标有字母的均值有显著差异(P<0.05)。
从图15得出的结论和注释:
25-OH-D3+抗氧化剂/抗炎剂改善自由采食饲喂的母鸡的慢性系统性炎症。
图16示出了饮食补充25-OH-D3+抗氧化剂/抗炎剂对具有限制性采食量或自由采食量的肉种母鸡的心肌细胞凋亡的影响(在35周龄时)。结果以均值±SEM(n=3)表示。条形上方标有字母的均值有显著差异(P<0.05)。
从图16得出的结论和注释:
25-OH-D3+抗氧化剂/抗炎剂改善具有限制性采食量或自由采食量的母鸡的心脏细胞凋亡。
结论:
补充的25-OH D3和抗氧化剂/抗炎剂通过以下方式来改善与肉种雌鸡过度饲喂有关的有害效应:
●降低死亡率并改善卵巢功能,从而降低过度饲喂的肉种母鸡的繁殖性能。
●改善内分泌(胰岛素)信号传导
●减少脂毒性发展和系统性炎症
●激活抗供能物过载诱导的心脏发病机理的心脏保护机制。
Claims (12)
1.一种适用于伴侣动物的食物,其包含组合,所述组合包含:25-羟基维生素D(25-OHD)、维生素C、维生素E以及类胡萝卜素,所述类胡萝卜素选自由以下各项组成的组:番茄红素、虾青素、隐黄质、β-胡萝卜素、叶黄素、玉米黄质和角黄素。
2.根据权利要求1的食物,其包含:25-OH D、β胡萝卜素、维生素C和维生素E,以及任选地还包含至少一种另外的生物活性成分,所述另外的生物活性成分选自由以下各项组成的组:维生素D、维生素B2、维生素B6、烟酸、锌、铜、锰和硒。
3.根据权利要求1或2的组合,其中维生素E或维生素C比β-胡萝卜素的量可以在40:1至1:1的范围内;优选地20:1至1:1;并且更优选地10:1至1:1。
4.根据权利要求1至3中任一项的组合,其中所述25-羟基维生素D是25-羟基维生素D3(25-OH D3)。
5.包含根据权利要求1至4中任一项的组合的饲料或根据权利要求1至4中任一项的饲料,其中:
25-OH D3的量为15至200μg/kg;
维生素E的量为40至400mg/kg;
抗坏血酸的量为40至400mg/kg;以及
β胡萝卜素的量为1至15mg/kg。
6.根据权利要求1至4中任一项的饲料,其中:
25-OH D3的量为80至150μg/kg;
维生素E的量为30至300mg/kg;
抗坏血酸的量为80至300mg/kg;以及
β胡萝卜素的量为1至15mg/kg。
7.根据权利要求1至4中任一项的饲料,其包含:
25-OH D3:35-150μg/kg;
维生素E:80-300mg/kg;
β胡萝卜素:3-12mg/kg;以及
抗坏血酸:100-300mg/kg。
8.根据权利要求1至4中任一项的饲料,其包含:
25-OH D3:35μg/kg
维生素E:80mg/kg
β-胡萝卜素:3mg/kg;以及
抗坏血酸:100mg/kg。
9.根据权利要求1至4中任一项的饲料,其包含:
25-OH D3:69μg/kg
维生素E:150mg/kg
β胡萝卜素:6mg/kg;以及
抗坏血酸:150mg/kg。
10.根据权利要求1至4中任一项的饲料,其包含:
25-OH D3:150μg/kg
维生素E:300mg/kg
β胡萝卜素:12mg/kg,以及
抗坏血酸:300mg/kg。
11.一种改善自由采食饲喂的伴侣动物的增加的体重的量的方法,其包括:将25-OH D、类胡萝卜素、维生素C和维生素E的组合施用于所述动物。
12.根据权利要求11的方法,其还包括:施用至少一种另外的生物活性成分,所述另外的生物活性成分选自由以下各项组成的组:
维生素D、维生素B2、维生素B6、烟酸、锌、铜、锰、硒,以及它们的组合。
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CN108935929A (zh) * | 2018-06-26 | 2018-12-07 | 辽宁禾丰牧业股份有限公司 | 肉鸡复合预混料及其制备方法 |
US11723919B2 (en) | 2020-06-12 | 2023-08-15 | Phibro Animal Health Corporation | Composition or combination comprising anionic dietary supplement and 25-hydroxy vitamin D |
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