CN1882351A - 组合物与方法 - Google Patents

组合物与方法 Download PDF

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CN1882351A
CN1882351A CNA2004800344456A CN200480034445A CN1882351A CN 1882351 A CN1882351 A CN 1882351A CN A2004800344456 A CNA2004800344456 A CN A2004800344456A CN 200480034445 A CN200480034445 A CN 200480034445A CN 1882351 A CN1882351 A CN 1882351A
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desulfovibrio
spp
gastrointestinal tract
helicobacter
cat
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C·古
K·L·格罗斯
G·吉布森
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Hills Pet Nutrition Inc
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Abstract

一种减少陪伴宠物胃肠道中脱硫弧菌和/或螺杆菌spp.数量的方法，包括给所述宠物口服施用脱硫弧菌和/或螺杆菌spp.减少量的纤维或其他组分。

Description

组合物与方法
发明背景
维护哺乳动物胃肠(GI)道的健康是非常期望的目标。与此特别相关的是胃肠道炎症。 脱硫弧菌(Desulfovibrio)spp.(包括但不限于 desulfuricans， intestinalis， vulgaris等)是硫酸盐还原菌，可以产生硫化氢，当由细菌释放时，能导致胃肠道细胞发炎。 螺杆菌 (helicobacter)(包括但不限于 heilmannii， felix， pylori，bizzozeronii， salomonis)能导致胃和肠上部细胞溃疡和发炎。胃肠道炎症体征包括急性或慢性腹泻、大便稀薄、血便、呕吐、营养消化和吸收不良、体重减轻和食欲不振。疾病，诸如胃炎、肠炎、炎性肠病、溃疡、某些类型的癌症以及其它一些疾病，已知是以胃肠炎症为主症的。将硫酸盐还原为硫化物的致病菌，诸如 脱硫弧菌spp，因为大便中硫化物或其他臭味化合物水平的增高，也能导致气体或大便臭味的增加。
我们现在已经发现，患有炎性肠病(IBD)的猫，比正常健康的猫， 脱硫弧菌和/或 螺杆菌spp.数量要高。我们还发现，螺杆菌在所有患有炎性肠病(IBD)的猫中都可以检测到，然而只有5/12受试的正常的猫检测到了螺杆菌。我们还发现，45％受试的IBD猫，双歧杆菌，一种有益菌群，的水平在标准检测水平以下，但是9％正常健康的猫，双歧杆菌在标准检测水平以下。
发明概述
根据本发明，提供了一种陪伴动物使用的可口服食用的食物组合物，包括可食用的食物组合物，结合以减少陪伴动物 脱硫弧菌和/或 螺杆菌spp.水平的组分。
本发明的另一方面，提供了一种减少陪伴宠物 脱硫弧菌和/或 螺杆菌spp.水平的方法，包括口服施用本发明的食物。
发明的详细说明
如前所述，现在已经发现，患有胃肠道炎症疾病IBD的猫，比没有此病的正常的猫， 脱硫弧菌spp.水平要高。因此，对于任何具有更高水平 脱硫弧菌和/或 螺杆菌spp.的陪伴宠物，无论它们有或没有一般伴随胃肠道炎症的疾病或病症的明显临床体征，使这种水平降低将都是有利的。阻止 脱硫弧菌和/或 螺杆菌spp.水平的升高同样有利，那是预防性作用。
活性剂可以减少所述细菌。这些活性剂包括抗菌材料，例如抗生素、化学治疗药物等。令人惊讶的是，纤维也能够降低 脱硫弧菌和/或螺杆菌spp.的水平。这样的纤维实例包括低聚糖、半乳聚糖、β-葡聚糖及其混合物。低聚糖实例包括低聚木糖，半乳低聚糖、低聚果糖等。β-葡聚糖的实例包括酵母细胞提取物、发芽大麦、燕麦纤维、凝胶多糖(curdlan)(微生物发酵得到的多糖)等。半乳聚糖的实例包括阿拉伯半乳聚糖等。优选地，多酚也可以和活性剂一起出现，尤其当活性剂是纤维，更尤其纤维是半乳聚糖，例如阿拉伯半乳聚糖时。多酚一般具有至少两个酚的结构，更优选黄酮类例如紫杉叶素。基于陪伴宠物的每日饮食，组合物中多酚最小量是大约0.01、0.05或0.1wt％。最高量一般不超过大约2、1、或0.75wt％，以陪伴宠物的每日饮食为准测量，所有重量都是以干物质为基础的。
也可使用抗 脱硫弧菌和/或 螺杆菌spp有效数量的组分。抗菌剂例如抗生素或化疗剂可以口服提供给宠物，最小量大约2 & 5mg/kg体重。最大量一般不多于大约25、50mg/kg体重。就纤维而言，以陪伴宠物每日饮食为准测量，最小量大约0.1、0.5或1.0wt％，最大量一般不应该超过大约5、10或20wt％，以干物质为基础称重。
脱硫弧菌和/或 螺杆菌spp.的减少能有效帮助处理以胃肠道炎症为主症(main component)的陪伴宠物的疾病和病症。陪伴宠物的实例是狗、猫、马，等。
实施例-显示IBD猫中提高水平的致病菌的存在.
1.筛选猫粪便样本的方案：
粪便样本收集自正常健康的猫和诊断为IBD的猫。正常猫以Science DietFeline maintenancedry维持，而IBD猫以治疗性胃肠饮食维持。粪便样本分析前-70℃冷冻。分析时，将样本和磷酸盐缓冲液以1∶10(w/w)的比例混合，玻璃珠涡旋，离心除去颗粒物。将375μl等分样本加入含有1.125ml 4％多聚甲醛的试管，4℃下留置4-5小时。将样本离心，在PBS中洗涤两次，然后和150μl滤过乙醇混合，-20℃冷藏，随后荧光原位杂交分析(用于微生物计数的FISH)。合成属特异性16S rRNA-靶向探针，在5′端用荧光染料单标记，以检测在发酵培养基中的相关细菌。将总核酸染色以获得总细胞计数。数据表示为log10细胞/g粪便。FISH可以对保藏的样本进行细菌定量，包括可培养的和非可培养的多样性。
结果
表1.IBD猫和正常猫致病菌菌落形成单位的Log10
                     正常猫                IBD猫
脱硫弧菌             cfu/g 7.0±2.5        7.7±0.6
粪便
螺杆菌               cfu/g 2.9±3.6        7.3±0.6
粪便
这些结果显示患胃肠道炎症，特别是IBD的猫，胃肠道出现的致病菌数量增加。
实施例2IBD猫和正常猫中AG对脱硫弧菌的体内作用
2.喂养研究方案
11只患有IBD的猫和10只正常健康的猫，以包含1.0％甜菜浆(beetpulp)和0.6％来自西部落叶松树的阿拉伯半乳聚糖提取物的食物喂养。所述的提取物是大约90wt％阿拉伯半乳聚糖和大约4wt％多酚，主要的多酚是紫杉叶素，剩余物是水分，都在干物质基础上，保持2周。随后，这些猫改用仅包含1.5％甜菜浆的食物喂养。在第0天，第14天和第28天收集粪便样本。按照下述方法为FISH分析制备样本：冷冻每份粪便样本，将5g粪便悬浮于在无菌袋或塑料容器中的pH 7.3的厌氧磷酸盐缓冲盐水(PBS)中，得到的终浓度为10％(5g 45ml)。淤浆在袋中匀质/混合以避免污染。每一样本使用不同容器。5ml淤浆和等量甘油混合，得到50∶50的混合物，冷冻以备FISH分析。
结果：
获得13套完整粪便样本。当所述的猫用含有0.6％AG提取物的食物喂养时，4/13的猫的脱硫弧菌 spp下降了0.3个对数单位和更高。8/13的猫的脱硫弧菌spp.水平下降很小或没有改变，而只有1/13的猫的脱硫弧菌spp.水平增加。当猫改用不含有AG的食物喂养时，10/13的猫脱硫弧菌spp.水平增加了0.3个对数单位和更高，2/13的猫没有改变，只有1/13的猫脱硫弧菌spp.水平下降。
结果显示AG提取物能阻止大多数的猫脱硫弧菌spp.水平增长，并使一些猫的脱硫弧菌spp.水平倾向于下降。这是在跟甜菜浆喂养的猫的水平的比较，而甜菜浆喂养导致大多数猫的脱硫弧菌spp水平增长。
实施例3-体外实验显示不同纤维降低 脱硫弧菌spp.的水平。
准备包含厌氧磷酸盐缓冲培养基的发酵容器，加入1ml犬粪便接种物(10％w/v粪便样本比缓冲液)。培养基组成见Sunvold GD，Hussein HS，Fahey GC，Merchen NR，和Reinhart GA(1995)，Invitro fermentation of selected fiber sources by dogs fecal inoculum andin vivo digestion and metabolism of fiber supplemental diets.J.AnimalSci.73：1099-1109(1995)的描述。发酵在39℃下进行。实验用不同的纤维以盲法-编码的方法进行。8小时孵育后，除去1ml培养液。制备它的一等分试样用于FISH分析。8小时后，除去1ml培养液，与在PBS中的4％多聚甲醛混合，固定用于FISH。合成属特异性16SrRNA-靶向探针，在5′端用荧光染料单标记，以检测在发酵培养基中的相关细菌。将总核酸染色以获得总微生物计数。结果显示8小时的发酵期间，几种不同类型的纤维能降低 脱硫弧菌spp.的生长0.5-1.0个对数单位(见表2)。
表2
8小时孵育后 脱硫弧菌spp.的数目(log cfu/ml粪便接种物).
 0小时时Log10CFU   8小时时LOG10CFU
  阿拉伯半乳聚糖  7.5±0.3   6.4±1.0
  低聚木糖  7.2±0.4   6.8±0.9
  半乳寡聚糖  7.0±1.0   6.8±0.9
  低聚果糖  6.9±1.0   6.3±0.8
  菊粉  7.3±0.2   6.4±1.0
  发芽大麦  6.8±0.9   5.8±0.0
总结
因此，我们已显示了在体外和体内AG降低脱硫弧菌spp.的水平。

Claims (10)

1.一种减少陪伴宠物胃肠道中的脱硫弧菌和/或螺杆菌spp.数量的方法，包括给所述宠物口服施用脱硫弧菌和/或螺杆菌spp.减少量(reducing quantity)的纤维，所述纤维选自低聚糖、半乳聚糖、β-葡聚糖及其混合物。
2.根据权利要求1的方法，其中所述陪伴宠物需要所述的施用。
3.根据权利要求2的方法，其中所述陪伴宠物是狗或猫。
4.根据权利要求3的方法，其中所述狗或猫患有其中以胃肠道炎症为主症的疾病。
5.根据权利要求4的方法，其中所述纤维选自阿拉伯半乳聚糖、低聚木糖、半乳低聚糖、低聚果糖、菊粉、发芽大麦及其混合物。
6.根据权利要求1的方法，其中也存在多酚。
7.一种治疗在胃肠道中具有提高水平的 脱硫弧菌和/或 螺杆菌 spp.的陪伴宠物中的胃肠道炎症的方法，包括口服施用减少 脱硫弧菌和/或 螺杆菌spp.有效量(reducing effective amount)的纤维。
8.根据权利要求7的方法，其中也存在多酚。
9.一种治疗在胃肠道中具有提高水平的 脱硫弧菌和/或 螺杆菌 spp.的陪伴宠物中的胃肠道炎症的方法，包括口服施用减少 脱硫弧菌有效量的组分，所述组分使胃肠道中 脱硫弧菌和/或 螺杆菌spp.的数量减少。
10.一种减少具有提高水平的 脱硫弧菌和/或 螺杆菌spp.的陪伴宠物气味的方法，所述气味选自肠气体气味、粪便气味和其任意混合，包括口服施用减少 脱硫弧菌和/或 螺杆菌spp.有效量的组分，所述组分使胃肠道中 脱硫弧菌和/或 螺杆菌spp.的数量减少。
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