CN111357878B - 一种减缓水牛热应激添加剂及其应用 - Google Patents
一种减缓水牛热应激添加剂及其应用 Download PDFInfo
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Abstract
本发明涉及热应激添加剂技术领域,特别涉及一种减缓水牛热应激的添加剂及其应用,本申请通过设计热应激环境,在水牛热应激条件下对其饲料中各营养成分表观消化率进行研究,评估日粮中添加烟酸铬对热应激泌乳水牛瘤胃发酵、泌乳性能、乳脂肪酸组成、抗氧化性能和细菌多样性的影响探究,从提高水牛瘤胃微生物多样性的角度探寻适宜的减缓水牛热应激添加剂,最终发现,适宜的烟酸铬添加量可增加水牛瘤胃的生物多样性,提高瘤胃微生物对养分的表观消化率,改善水牛抗热应激效果,使水牛在热应激条件下仍然保有较高的瘤胃消化能力和饲料转化效率,为烟酸铬在水牛生产上的应用提供科学依据。
Description
【技术领域】
本发明涉及减缓热应激添加剂技术领域,特别涉及一种减缓水牛热应激添加剂及其应用。
【背景技术】
水牛是热带、亚热带地区特有的家畜,具有乳、肉、役的多种用途,有着较高的经济价值。由于水牛能够很好地适应泥泞沼泽地的湿热气候条件,因此在热带和亚热带地区能够长期存在。水牛还可将低质粗饲料有效地转换成为乳、肉等高附加值的产品,从而减少农作物副产物等废弃物对环境的影响。此外,与黄牛相比,水牛乳含有较高的蛋白质、脂肪和非脂固形物。然而,由于气候变化导致的全球变暖,温度,湿度和辐射的变化会对家畜物种的生长和生产带来潜在威胁。随着全球变暖趋势加剧,特别是热带亚热带地区在随后几十年内平均气温将会升高较快,高温环境是对生活在热带和亚热带地区水牛生产力的主要约束因子。预计到2050年,温度将会再增加1~3℃,极端高温事件愈加频繁,气候变暖正逐渐成为热带与亚热带地区奶业发展的限制性因素。当动物遭受热应激时,会导致干物质摄入量减少,新陈代谢减慢,甲状腺功能减退,这会直接影响生长、泌乳性能、繁殖效率和饲料消化率,导致相当大的经济损失,特别是水牛,因为水牛皮肤呈黑色,被毛稀疏,汗腺密度低,在高温环境下,泌乳量、生长速度和繁殖性能都受到了极大的限制。一般认为温湿指数(temperature humidityindex;THI)超过89时水牛处于严重热应激状态,本课题组前期研究发现,广西南宁6-9月份THI平均值达80以上,水牛呼吸频率及体表温度均明显升高,瘤胃发酵模式改变,养分摄入量下降等热应激现象显著,导致水牛经济性能受到严重影响。
因此,有必要在热应激条件下针对水牛的生理特点研制出一种适合水牛特性的减缓热应激的添加剂,并应用到水牛中,以提高水牛的生长和泌乳性能。
【发明内容】
本发明的目的是提供一种减缓水牛热应激添加剂及其应用,该添加剂能提高水牛瘤胃微生物的丰度,从根源上提高水牛在热应激条件下对饲料的消化、吸收能力,提高养分的摄入量,从而达到提高产奶量,改善肉质的目的。
一种减缓水牛热应激添加剂,所述添加剂包含烟酸铬。
本申请还包括一种应用所述减缓水牛热应激添加剂在提高水牛产奶量上的饲喂方法,所述方法是在基础日粮中添加3mg/(头·d)的烟酸铬。
本申请还包括所述减缓水牛热应激添加剂在提高水牛乳乳脂中C12:0和C14:0脂肪酸含量的应用。
本申请还包括一种应用所述减缓水牛热应激添加剂在提高水牛乳乳脂中C12:0和C14:0脂肪酸含量的饲喂方法,所述方法是在基础日粮中添加3毫克/头/天的烟酸铬。
本申请还包括所述减缓水牛热应激添加剂在提高水牛瘤胃微生物群落多样性上的应用。
本申请还包括一种应用所述减缓水牛热应激添加剂在提高反刍动物瘤胃微生物群落多样性上的饲喂方法,所述方法是在基础日粮中添加1.5-3毫克/头/天的烟酸铬。
进一步的,所述基础日粮由如下重量百分数的成分组成:32.82%象草,48.28%啤酒糟,9.44%玉米,3.98%小麦麸,3.03%豆粕,0.26%石粉,0.37%食盐,0.64%磷酸氢钙,预混料1.18%。
进一步的,所述预混料中每千克饲粮中含如下配比的微量元素:维生素A550000IU,维生素E3000IU,维生素D3150000IU,硫酸亚铁4.0g、硫酸铜1.3g,硫酸锰3.0g,硫酸锌6.0g,硫酸钴80mg。
进一步的,所述奶水牛喂养期,试验期牛舍的平均温度为31.95℃,平均温湿指数为85.32。
本发明具有以下有益效果:
本申请通过设计热应激环境,在水牛热应激条件下对其饲料中各营养成分表观消化率进行研究,评估日粮中添加抗氧化剂烟酸铬对热应激泌乳水牛瘤胃发酵、泌乳性能、乳脂肪酸组成、抗氧化性能以及细菌多样性的探究,从提高水牛瘤胃微生物多样性的角度探寻适宜的减缓水牛热应激添加剂,最终发现,适宜的烟酸铬添加量可增加水牛瘤胃的生物多样性,提高瘤胃微生物对养分的表观消化率,改善水牛抗热应激效果,使水牛在热应激条件下仍然保有较高的瘤胃消化能力和饲料转化效率,为烟酸铬在水牛生产上的应用提供科学依据。
【附图说明】
图1为各组水牛瘤胃液菌群物种分布Venn图;
图2为门水平上瘤胃细菌群落丰度百分比图;
图3为属水平上瘤胃细菌群落丰度百分比图;
图4为瘤胃微生物PCA分析图。
【具体实施方式】
下面结合附图和实施例试验对本发明作进一步说明。
实施例1:
1、试验动物及地点
试验于广西水牛研究所种畜场进行,选取泌乳量接近泌乳中期奶水牛20头,随机分为4组,每组5只。试验周期5周,预饲期1周,正饲期4周,工人饲喂,正常生产。试验期牛舍的平均温度为31.95℃,平均温湿指数(THI)为85.32,已知当THI在72~79范围时为轻度热应激期,79~88为中度热应激,89以上为严重热应激。在整个试验期间,水牛都处于热应激状态。
基础日粮采用牛场泌乳牛日粮见表1,试验组饲喂分别添加1.5、3和4.5毫克/头/天烟酸铬(市售,烟酸铬≥1.65%,铬≥0.2%)的试验饲粮。
表1泌乳期水牛基础饲粮组成及营养水平(风干基础)%
上表中预混料为每千克饲粮中含如下配比的微量元素:维生素A550000IU,维生素E3000IU,维生素D3150000IU,硫酸亚铁4.0g、硫酸铜1.3g,硫酸锰3.0g,硫酸锌6.0g,硫酸钴80mg。
2、试验材料
烟酸铬添加剂为市售饲料添加剂,烟酸铬≥1.65%,铬≥0.2%
3、检测指标
①采食量及表观消化率
试验最后一周连续三天检测水牛饲喂量、剩料量,计算干物质采食量;采集精料及粗料样品和粪样测定干物质、粗灰分、粗蛋白、中性洗涤纤维、酸性洗涤纤维等含量,计算表观消化率。
②瘤胃发酵参数及微生物DNA提取
试验最后一天晨饲前,从每组试验牛中随机选取3头,通过胃管式瘤胃液采样器经口腔采集瘤胃液,装入瓶中置于冰盒中保存,迅速带回实验室测定pH,并按照瘤胃液指标的需要合理分装并适当保存,以备测定各项瘤胃液发酵参数。瘤胃液pH采用酸度计测定。瘤胃液挥发性脂肪酸(VFA)浓度采用气相色谱仪(Aglient7890B,美国)测定;氨态氮(NH3-N)浓度采用苯酚-次氯酸钠比色法测定;微生物蛋白(MCP)浓度采用考马斯亮蓝法测定。取1.0mL瘤胃微生物参照Wright苯酚-氯仿-异戊醇方法抽提瘤胃内容物总DNA,DNA浓度用NanoDrop测定,送美吉生物公司进行16SrDNA高通量测序。
③产奶性能及乳品质
试验期间记录每天每头水牛的产奶量,计算日均产奶量;在试验期内每周三上午、下午各取50mL奶样,分别使用FOSS全自动乳成分分析仪进行乳成分测定,包括乳脂肪、乳蛋白质、乳糖、乳密度、乳脂固形物等。将最后一周采集的奶样按上午30mL、下午20mL混匀置于-20℃保存,送乳脂脂肪酸组成及含量检测。
4%乳脂校正乳产量(kg/d)=0.4×乳产量(kg/d)+0.15×乳脂率×乳产量(kg/d)
常乳泌乳效率(%)=产奶量(kg/d)/干物质采食量(kg/d)
④血清抗氧化指标
试验最后一天晨饲前颈静脉采集血液,离心分离血清,-20℃以下保存备测抗氧化相关指标。测定指标包括:总超氧化物歧化酶(T-SOD)、总抗氧化能力(T-AOC)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)与过氧化氢酶(CAT)活性。以上指标测试试剂盒均购于南京建成生物工程研究所,测定方法按照试剂盒操作说明书进行。
5、数据统计分析
试验数据采用SPSS19.0软件进行单因素方差分析,差异显著者采用Duncan氏多重比较检验,以P<0.05为差异显著。试验数据用平均值±标准差表示。
6、实验结果:
①烟酸铬对水牛采食量及养分表观消化率的影响(见表2):
表2
由上表知,饲粮中添加烟酸铬可提高水牛DMI、ADF和NDF的表观消化率,但差异不显著(P>0.05);与Ⅰ组相比,Ⅲ组和Ⅳ组干物质采食量和CP表观消化率提高,而Ⅱ组的均下降,差异不显著(P>0.05)。本试验中,干物质采食量差异不显著,添加不同水平的烟酸铬ADF和NDF的表观消化率均有不同程度的上升。
②烟酸铬对水牛瘤胃发酵参数的影响(见表3):
表3
由上表知,与Ⅰ组相比,Ⅱ、Ⅲ和Ⅳ组的瘤胃乙酸、丙酸、丁酸、等挥发性脂肪酸无显著影响(P>0.05);Ⅱ组的氨态氮和微生物蛋白较Ⅰ组有所下降,Ⅲ组和Ⅳ组的均有所提高,差异都不显著(P>0.05);各组水牛的瘤胃pH在6.53~6.80范围内,试验组与对照组无显著差异(P>0.05);本试验中,试验组丙酸和丁酸浓度与对照组相比均有所提高,但差异均不显著,添加烟酸铬使瘤胃发酵类型往丙酸或丁酸发酵型转换。
③烟酸铬对水牛泌乳性能的影响(见表4):
表4
由上表知,与Ⅰ组相比,Ⅲ组的水牛产奶量显著提高了7.70%(P<0.05),Ⅱ、Ⅲ和Ⅳ组乳蛋白、乳脂、乳总固形物和乳非脂固形物率较Ⅰ组差异不显著(P>0.05),但乳蛋白有不同程度的升高;Ⅲ和Ⅳ组的4%乳脂校正乳产量显著高于Ⅰ组(P<0.05),分别提高了7.5%和8.3%;与Ⅰ组比较,Ⅱ组的常乳泌乳效率显著提高(P<0.05)。
④烟酸铬对水牛减缓氧化性能的影响(见表5):
表5
由上表知,与Ⅰ组相比,Ⅱ、Ⅲ和Ⅳ组水牛血清过氧化氢酶(CAT)不同程度的下降,无显著差异(P>0.05);Ⅱ、Ⅲ和Ⅳ组的总超氧化物歧化酶(T-SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-PX)与Ⅰ组差异不显著(P>0.05),但Ⅱ、Ⅲ组的T-SOD显著高于Ⅳ组(P<0.05),Ⅱ组的MDA显著高于Ⅲ组(P<0.05);与Ⅰ组相比,Ⅲ组的和T3和T4浓度显著增加(P<0.05)。
⑤中、长链脂肪酸分析结果(见表6):
表6
从上表可知,Ⅲ组的C14:1n5、C16:1n7的含量显著高于Ⅰ组(P<0.05);Ⅲ组C12:0和C14:0的含量显著高于Ⅳ组(P<0.05);数据显示日粮中添加烟酸铬后三个试验组(Ⅱ、Ⅲ、Ⅳ)乳脂中C18:2(cis-9,trans-11)、C20:2n6、C20:5n3(EPA)、C22:6n3(DHA)和C24:0的含量较Ⅰ组均有不同程度的提高,但无显著差异(P>0.05),说明了动物日粮中添加抗氧化剂也可以保护不饱和脂肪酸不被氧化,从而增加水牛乳中不饱和脂肪酸含量,而相对减少饱和脂肪酸的含量。
⑥瘤胃细菌多样性结果(参见图1-图4):
利用细菌群落中共享和独有的细菌丰度对各组水牛瘤胃微生物进行分析,如图1所示。花瓣图代表水牛瘤胃液各组样本的物种共有和独有分类操作单元(OTU)分布情况,如图1可知,不同色块代表不同组别,其中,CN1为对照组(即不在日粮中添加烟酸铬);CN2为低剂量组(即在日粮中添加1.5毫克/头/天的烟酸铬);CN3为中剂量组(即在日粮中添加3.0毫克/头/天的烟酸铬);CN4为高剂量组(即在日粮中添加4.5毫克/头/天的烟酸铬);在97%相似性水平下,12牛水牛瘤胃液样本生成2804个OTU,其中对照组(CN1)的OTU为2218,低剂量试验组(CN2)OTU为2213,中剂量试验组(CN3)OTU为2252,高剂量试验组(CN4)OTU为2173个。其中对照组(CN1)、添加1.5毫克/头/天烟酸铬组(CN2)、添加3毫克/头/天烟酸铬组(CN3)和添加4.5毫克/头/天烟酸铬组(CN4)组独有的OTU数分别占OTU总数的3.07%、3.21%、4.53%和2.46%。而添加3毫克/头/天烟酸铬组比对照组OTU数目增多,4.5毫克/头/天烟酸铬组的OTU数目则降低。
说明添加烟酸铬对瘤胃微生物多样性是有一定影响的,菌落种类有所增加,但到了较高剂量后,其微生物水平反而呈现下降趋势,这说明烟酸铬的添加量并非越多越好,而且添加量是不能通过推理预期的。
图2为门水平上瘤胃细菌群落丰度百分比,其中,(A1、A2、A3)为对照组(0毫克/头/天);(A4、A5、A6)为低剂量烟酸铬组(1.5毫克/头/天);(A7、A8、A9)为中剂量烟酸铬组(3.0毫克/头/天);(A10、A11、A12)为高剂量烟酸铬组(4.5毫克/头/天)。
由图2可知,在门水平,将在所有样本中丰度占比均小于0.01的物种归为others,其菌群主要属于7个门:Bacteroidetes、Firmicutes、Proteobacteria),Cyanobacteria、Spirochaetes、Patescibacteria、Kiritimatiellaeota。各组的优势菌群为拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)变形菌门(Proteobacteria),这三类菌占总细菌比例的90%左右。不同处理组之间菌群丰富度存在差异,其中1.5和3mg/天/头烟酸铬组的拟杆菌门数目较对照组有所提高,而厚壁菌门的数目较对照组降低。
图3为属水平上瘤胃细菌群落丰度百分比,其中,(A1、A2、A3)为对照组;(A4、A5、A6)为低剂量烟酸铬组(1.5毫克/头/天);(A7、A8、A9)为中剂量烟酸铬组(3.0毫克/头/天);(A10、A11、A12)为高剂量烟酸铬组(毫克/头/天)。
从图3可见,在属水平上,将在所有样本中丰度占比小于0.02的物种归为others时,由图3群落柱形图可知,瘤胃微生物由普雷沃氏菌属Prevotella、克里斯滕森菌科R-7群ChristensenellaceaeR-7group、理研菌科RC9肠道群RikenellaceaeRC9gutgroup、疣微菌科NK4A214群RuminococcaceaeNK4A214group等17个属组成。很明显,不同样本其微生物组成大至相似,只是占比有所不同,样本整体呈现出较丰富的多样性。因此,在泌乳奶水牛日粮中添加高剂量烟酸铬对瘤胃微生物影响不明显。
图4为各组瘤胃液落菌PCA分析图,其中CN1(A1、A2、A3)为对照组;CN2(A4、A5、A6)为低剂量烟酸铬组(1.5毫克/头/天);CN3(A7、A8、A9)为中剂量烟酸铬组(3.0毫克/头/天);CN4(A10、A11、A12)为高剂量烟酸铬组(4.5毫克/头/天)。
由图4可知,各组间的菌落构成有一定的差异,尤其是饲喂了1.5毫克/头/天烟酸铬组(CN2),与对照组相(CN1)比,其样本核心微生物组成点差距甚远,说明无相似物种,这可能是刚开始添加烟酸铬的时候,个体需要一定的适应性。饲喂了3mg烟酸铬/头/天组(CN3)和4.5毫克/头/天烟酸铬组(CN4)相比,样本点与点之间的差距相对较近,有聚类现象,说明其物种组成较相似。由此可见,在泌乳奶水牛基础日粮中添加烟酸铬对瘤胃微生物影响较显著。
反刍动物主要通过瘤胃内的微生物发酵来消化食物,瘤胃是反刍动物的前胃,微生物密度大,种类多,菌群对宿主的正常功能和发育至关重要。据研究报道,反刍动物瘤胃内的主要微生物菌群在门水平上是拟杆菌门和厚壁菌门,与本试验结果相一致。从图1-4可以看出,基于门水平的比较,其中拟杆菌门数目的提高,会影响厚壁菌门占总菌群的比例,这可能会影响瘤胃微生物主要功能的转移。本试验中,普氏菌属均是存在于瘤胃液中丰度最高的菌属。这说明,日粮组成不仅影响反刍动物瘤胃微生物区系的组成,而且在其整个生长过程中,随着日粮组成的改变,其瘤胃微生物物种分布也在动态改变。
综上,本申请的抗水牛热应激添加剂应用于奶水牛日粮中,能有效缓解水牛热应激,影响瘤胃微生物群落的多样性组成,同时可促进挥发性脂肪酸(VFAs)和微生物蛋白的产生,提高奶水牛产奶量和肉质,是一种安全、可靠的减缓水牛热应激添加剂,其最佳添加量为3毫克/头/天。
上述说明是针对本发明较佳可行实施例的详细说明,但实施例并非用以限定本发明的专利申请范围,凡在本发明所提示的技术精神下所完成的同等变化或修饰变更,均应属于本发明所涵盖专利范围。
Claims (7)
1.减缓水牛热应激添加剂在提高水牛乳乳脂中C12:0和C14:0脂肪酸含量上的应用,其特征在于,所述添加剂包含烟酸铬,所述烟酸铬在基础日粮中的添加量为3毫克/头/天。
2.一种应用权利要求1所述减缓水牛热应激添加剂在提高水牛乳乳脂中C12:0和C14:0脂肪酸含量的饲喂方法,其特征在于,所述方法是在基础日粮中添加3毫克/头/天的烟酸铬。
3.减缓水牛热应激添加剂在提高水牛瘤胃微生物群落多样性上的应用,其特征在于,所述添加剂包含烟酸铬,所述烟酸铬在基础日粮中的添加量为3毫克/头/天。
4.一种应用权利要求3所述减缓水牛热应激添加剂在提高反刍动物瘤胃微生物多样性上的饲喂方法,其特征在于,所述方法是在基础日粮中添加3毫克/头/天的烟酸铬。
5.根据权利要求2或4所述的饲喂方法,其特征在于,所述基础日粮由以下重量百分数的组成:32.82%象草,48.28%啤酒糟,9.44%玉米,3.98%小麦麸,3.03%豆粕,0.26%石粉,0.37%食盐,0.64%磷酸氢钙,预混料1.18%。
6.根据权利要求5所述的饲喂方法,所述预混料中每千克饲粮中含如下配比的微量元素:维生素A 550000IU,维生素E 3000 IU,维生素D 3150000IU,硫酸亚铁4.0g、硫酸铜1.3g,硫酸锰3.0g,硫酸锌6.0g,硫酸钴80mg。
7.根据权利要求2或4所述的饲喂方法,其特征在于,所述水牛的喂养期,牛舍的平均温度为31.95℃,平均温湿指数为85.32。
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JP2006000061A (ja) * | 2004-06-18 | 2006-01-05 | Marubeni Nisshin Feed Co Ltd | 家禽飼料用添加剤及び家禽飼料 |
CN1823608A (zh) * | 2006-03-29 | 2006-08-30 | 浙江大学 | 控缓释型铬的补铬饲料添加剂及其制备与使用方法 |
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US6139881A (en) * | 1999-02-09 | 2000-10-31 | Thompson Animal Systems, Inc. | Chromium-carboxylic acid feed supplement |
CN1618316A (zh) * | 2003-11-19 | 2005-05-25 | 福建农林大学 | 奶牛热应激缓解剂及其制备方法和应用 |
JP2006000061A (ja) * | 2004-06-18 | 2006-01-05 | Marubeni Nisshin Feed Co Ltd | 家禽飼料用添加剤及び家禽飼料 |
CN1823608A (zh) * | 2006-03-29 | 2006-08-30 | 浙江大学 | 控缓释型铬的补铬饲料添加剂及其制备与使用方法 |
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