动物营养学报    2021, Vol. 33 Issue (7): 3896-3903    PDF    
青蒿提取物对奶牛泌乳性能、血浆免疫与抗氧化指标的影响
余诗强1 , 熊安然1 , 潘予琮1 , 张亚静2 , 汪悦3 , 蒋林树1 , 熊本海3     
1. 北京农学院动物科学技术学院, 奶牛营养学北京市重点实验室, 北京 102206;
2. 河北省高碑店市职教中心, 保定 074000;
3. 中国农业科学院北京畜牧兽医研究所, 北京 100193
摘要: 本试验旨在研究青蒿提取物对奶牛泌乳性能、血浆免疫与抗氧化指标的影响。采用完全随机区组设计,将20头中国荷斯坦奶牛按照泌乳期、体重、产奶量和胎次等相似原则随机分为4组,每组5头。对照组饲喂基础饲粮,试验组在基础饲粮中分别添加50、100、150 g/(d·头)的青蒿提取物。预试期10 d,正试期35 d。结果表明:1)与对照组相比,添加150 g/(d·头)青蒿提取物显著提高了产奶量和乳糖率(P < 0.05);添加100 g/(d·头)青蒿提取物显著提高了4%乳脂校正乳产量和乳糖率;添加50 g/(d·头)青蒿提取物显著提高了乳糖率(P < 0.05)。2)与对照组相比,添加50、100、150 g/(d·头)青蒿提取物显著升高了血浆中免疫球蛋白M(IgM)的浓度(P < 0.05),显著降低了血浆中白细胞介素-1β(IL-1β)的浓度(P < 0.05),对血浆中免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、γ-干扰素(IFN-γ)的浓度无显著影响(P>0.05);血浆中白细胞介素-6(IL-6)的浓度在50和100 g/(d·头)的添加水平下没有显著变化(P>0.05),在150 g/(d·头)的添加水平下显著提高(P < 0.05)。3)与对照组相比,添加50、100、150 g/(d·头)青蒿提取物显著提高了血浆中超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)的活性(P < 0.05),显著降低了血浆中丙二醛(MDA)的浓度(P < 0.05),对血浆中过氧化氢酶(CAT)的活性无显著影响(P>0.05)。综上所述,饲粮中添加青蒿提取物能够提高奶牛的泌乳性能、机体免疫力和抗氧化性能。本试验条件下,奶牛饲粮中青蒿提取物的适宜添加水平为100 g/(d·头)。
关键词: 青蒿提取物    奶牛    泌乳性能    免疫指标    抗氧化指标    
Effects of Artemisia annua L. Extract on Lactation Performance, Plasma Immune and Antioxidant Indexes of Dairy Cows
YU Shiqiang1 , XIONG Anran1 , PAN Yucong1 , ZHANG Yajing2 , WANG Yue3 , JIANG Linshu1 , XIONG Benhai3     
1. Key Laboratory for Dairy Cow Nutrition, College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China;
2. Hebei Gaobeidian Vocational Education Center, Baoding 074000, China;
3. Beijing Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Abstract: The purpose of this experiment was to study the effects of Artemisia annua L. extract on the lactation performance, plasma immune and antioxidant indexes of dairy cows. A randomized complete block design was adopted in this experiment. Twenty Chinese Holstein dairy cows were randomly divided into 4 groups with 5 cows per group according to similar lactation period, body weight, milk yield and parity. Dairy cows in control group were fed a basal diet, while those in experimental groups were fed the basal diet supplemented with Artemisia annua L. extract of 50, 100 and 150 g/(d·head), respectively. The pre-test period was 10 days, and the test period was 35 days. The results showed as follows: 1) compared with the control group, adding 150 g/(d·head) Artemisia annua L. extract significantly increased the milk yield and lactose percentage of dairy cows (P < 0.05), adding 100 g/(d·head) Artemisia annua L. extract significantly increased the 4% fat corrective milk yield and lactose percentage of dairy cows (P < 0.05), and adding 50 g/(d·head) Artemisia annua L. extract significantly increased the lactose percentage of dairy cows (P < 0.05). 2) Compared with the control group, adding 50, 100 and 150 g/(d·head) Artemisia annua L. extract significantly increased the plasma immunoglobulin M (IgM) concentration (P < 0.05), and significantly decreased the plasma interleukin-1β (IL-1β) concentration (P < 0.05), but had no significant effects on the plasma immunoglobulin A (IgA), immunoglobulin G (IgG) and interferon-γ (IFN-γ) concentrations (P>0.05). The concentration of interleukin-6 (IL-6) in plasma did not change significantly at 50 and 100 g/(d·head) supplemental levels (P>0.05), but increased significantly at 150 g/(d·head) supplemental level (P < 0.05). 3) Compared with the control group, adding 50, 100 and 150 g/(d·head) Artemisia annua L. extract significantly increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in plasma (P < 0.05), significantly reduced the concentration of plasma malondialdehyde (MDA) (P < 0.05), but has no significant effect on the concentration of plasma catalase of dairy cows. To sum up, adding Artemisia annua L. extract in the diet can improve the lactation performance, immune and antioxidant capacity of dairy cows. Under the conditions of this experiment, the optimum adding level of Artemisia annua L. extract in the diet of dairy cows is 100 g/(d·head).
Key words: Artemisia annua L. extract    dairy cows    lactation performance    immune indexes    antioxidant indexes    

研制和开发安全、绿色和无残留等特性的新型饲料添加剂已经成为实现畜牧业健康养殖的重要技术途径[1]。天然植物提取物具有的抗菌、抗炎、抗氧化应激及提高动物饲料转化率等生物学功能,成为新型饲料添加剂研究热点[2-4]。青蒿(Artemisia annua L.),又名草蒿、香蒿、臭蒿等,是菊科一年生草本植物[5]。研究表明,青蒿提取物可以通过抑制白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的产生,阻止脂多糖(LPS)加速核转录因子-κB(NF-κB)中p65亚基向细胞核迁移,发挥保护细胞免受氧化应激和炎症损伤的作用[6]。青蒿中的活性成分能够以剂量依赖的方式显著抑制活化的中性粒细胞产生TNF-α,对环氧化酶炎症标记物前列腺素E2也具有显著抑制作用,抑制率可达93%[7]。侯伟[8]研究发现,将青蒿提取物与牛奶混匀后饲喂给犊牛,可以提高犊牛开食料采食量和日增重,降低腹泻率,并显著提高血液中总超氧化物歧化酶(T-SOD)活性以及免疫球蛋白A(IgA)、免疫球蛋白G(IgG)和可溶性CD4的浓度,提高犊牛机体的免疫水平。Jiao等[9]研究发现,青蒿提取物可通过降低大肠杆菌(Escherichia coli)引起的白细胞介素-17A(IL-17A)和NF-κB的高表达发挥抑菌和抗炎作用。因此,本试验拟探究饲粮中添加青蒿提取物对奶牛泌乳性能、血浆免疫与抗氧化指标的影响,以期为青蒿提取物在奶牛生产中的应用提供理论依据。

1 材料与方法 1.1 试验材料

本试验所用的青蒿提取物为棕黄色粉末,选购自陕西某公司,其主要成分和含量如下:青蒿素39.0%、粗灰分13.0%、粗纤维27.9%、粗蛋白质6.3%、水分5.0%、多糖8.3%、挥发油0.5%。

1.2 试验设计

选取20头体况相近、胎次为2~4胎、日产奶量为(33.0±3.9) kg的中国荷斯坦奶牛作为试验动物。采用完全随机区组设计,将20头奶牛按照泌乳期、体重、产奶量和胎次等相似原则随机分为4组,每组5头。对照组饲喂基础饲粮,试验组在基础饲粮中分别添加50、100、150 g/(d·头)的青蒿提取物。预试期10 d,正试期35 d。试验时间为2019年5—7月份,试验地点为奶牛营养学北京市重点实验室延庆基地。

1.3 基础饲粮与饲养管理

基础饲粮参照NRC(2001)奶牛饲养标准配制,其组成及营养水平由试验牛场提供,详见表 1。试验牛采用散栏带卧床式饲养,基础饲粮以全混合日粮(TMR)形式饲喂,青蒿提取物于每天晨饲前进行投喂,自由饮水,每天09:30、15:30、21:30进行挤奶。

表 1 基础饲粮组成及营养水平(干物质基础) Table 1 Composition and nutrient levels of the basal diet (DM basis) 
1.4 样品采集与指标测定 1.4.1 奶样采集与指标测定

产奶量测定:采用2×16系列并列式挤奶机记录每日产奶量。

乳成分测定:在正试期最后1 d分别采集各组奶牛早、中、晚的鲜奶样。挤奶前清洁乳房,弃去头3把奶,按照早、中、晚4 : 3 : 3的比例混合得到50 mL待测奶样,置于DHI专用样品瓶中,加入重铬酸钾作为奶样防腐剂,混合均匀后,于当天送至北京市清河奶牛中心进行乳成分分析。具体测定指标为:乳脂率、乳蛋白率、乳糖率及体细胞数。

4%乳脂校正乳产量和能量校正乳产量计算公式如下:

1.4.2 血样采集与指标测定

在正试期最后1 d于晨饲前对每头奶牛进行尾静脉采血,并将血液样本收集至含肝素钠抗凝的真空采血管中,室温静止30 min,以3 000×g、4 ℃离心15 min分离得到血浆,用离心管分装后于-20 ℃下保存待测。

血浆免疫指标包括:IgA、IgG、免疫球蛋白M(IgM)、IL-1β、白细胞介素-6(IL-6)、γ-干扰素(IFN-γ)的浓度。

血浆抗氧化指标包括:超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)的活性和丙二醛(MDA)的浓度。

血浆免疫与抗氧化指标均采用试剂盒测定,试剂盒由南京建成生物工程研究所提供,试剂配制及具体操作步骤按照说明书进行。

1.5 数据统计与分析

试验数据经Excel 2016进行初步整理后,采用SPSS 20.0统计软件进行方差分析,组间采用Duncan氏法进行多重比较,并以P < 0.05作为差异显著性判断标准。

2 结果 2.1 青蒿提取物对奶牛产奶量及乳成分的影响

表 2可知,与对照组相比,添加150 g/(d·头)青蒿提取物显著提高了产奶量和乳糖率(P < 0.05);添加100 g/(d·头)青蒿提取物显著提高了4%乳脂校正乳产量和乳糖率(P < 0.05);添加50 g/(d·头)青蒿提取物显著提高了乳糖率(P < 0.05);添加50、100和150 g/(d·头)青蒿提取物对乳脂率、乳蛋白率和体细胞数均无显著影响(P>0.05)。

表 2 青蒿提取物对奶牛产奶量及乳成分的影响 Table 2 Effects of Artemisia annua L. extract on milk production and milk composition of dairy cows
2.2 青蒿提取物对奶牛血浆免疫指标的影响

表 3可知,与对照组相比,添加50、100和150 g/(d·头)青蒿提取物对奶牛血浆中IgA、IgG、IFN-γ的浓度未产生显著影响(P>0.05),但显著升高了血浆中IgM的浓度(P < 0.05),显著降低了血浆中IL-1β的浓度(P < 0.05);血浆中IL-6的浓度在青蒿提取物添加水平为50和100 g/(d·头)时没有显著变化(P>0.05),在青蒿提取物添加水平为150 g/(d·头)时显著提高(P < 0.05)。

表 3 青蒿提取物对奶牛血浆免疫指标的影响 Table 3 Effects of Artemisia annua L. extract on plasma immune indexes of dairy cows
2.3 青蒿提取物对奶牛血浆抗氧化指标的影响

表 4可知,与对照组相比,添加50、100和150 g/(d·头)青蒿提取物显著降低了血浆中MDA的浓度(P < 0.05),对血浆中CAT的活性没有产生显著影响(P>0.05);添加50、100和150 g/(d·头)青蒿提取物显著提高了血浆中SOD的活性;添加150 g/(d·头)青蒿提取物显著提高了血浆中GSH-Px的活性(P < 0.05)。

表 4 青蒿提取物对奶牛血浆抗氧化指标的影响 Table 4 Effects of Artemisia annua L. extract on plasma antioxidant indexes of dairy cows
3 讨论 3.1 青蒿提取物对奶牛产奶量及乳成分的影响

本试验结果表明,在泌乳奶牛饲粮中添加青蒿提取物能提高奶牛的产奶量、乳糖率及4%乳脂校正乳产量,此结果与侯昆等[10]的研究结果相似。奶牛饲粮中添加植物提取物能提高奶牛的泌乳性能,植物提取物中的营养活性物质可以调节瘤胃的发酵功能及相关泌乳信号通路,促进脂肪酸合成,进而使乳成分发生改变[11-12]。饲喂奶牛青蒿提取物(含39%青蒿素)可以促进瘤胃脂肪酸的合成、提高乳腺上皮细胞泌乳能力,并通过激活磷酸化腺苷酸活化蛋白激酶(AMPK)-哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的转导,增加PPARγ、mTOR、固醇调节元件结合蛋白-1c(SREBP1-c)等蛋白的表达水平,提高乳脂的合成量,使奶牛的产奶量和4%乳脂校正乳产量显著提高[13]。另有研究表明,乳糖与其他乳成分相比受不确定因素的影响较小,产奶量与乳糖率存在着显著的正相关关系[14],与乳蛋白率和体细胞数呈显著的负相关关系,与乳脂率无显著相关性[15],这与本试验结果基本相同。青蒿提取物中富含的黄酮和萜类化合物可以改善瘤胃发酵,促进瘤胃短链脂肪酸的合成,提高有机物质的消化率,增加泌乳净能和代谢能[16]。但目前关于青蒿素对奶牛产奶量影响的报道有限。本试验中,在150 g/(d·头)添加水平下奶牛产奶量提高显著,在100 g/(d·头)添加水平下4%乳脂校正乳产量提高显著,说明这2个添加水平的青蒿提取物可以明显提高奶牛的泌乳性能。这可能是因为青蒿中富含的萜类和黄酮等营养活性物质改变了瘤胃发酵模式,促进了营养物质的消化与吸收,维持了机体的代谢平衡,进而提高了泌乳性能。

3.2 青蒿提取物对奶牛血浆免疫指标的影响

相关研究表明,青蒿中的倍半萜、二萜、黄酮和挥发油等成分在抗炎和免疫调节方面发挥着主要作用[17]。IgA、IgG和IgM是反映机体免疫水平的重要指标。据报道,在奶牛饲粮中添加富含黄酮、萜类的植物提取物后,奶牛血液中IgG和IgM的浓度有所增高但不显著,但奶牛的整体免疫水平得到提高[18],这与本研究结果一致。这可能是由于植物提取物中黄酮类、萜类物质通过重新调节免疫球蛋白的含量来影响体液免疫[19-20]。本试验中,奶牛血浆中IgA、IgG和IgM的浓度在添加青蒿提取物后均有不同程度的提高,表明青蒿提取物对奶牛机体免疫具有一定的促进作用,但具体是青蒿提取物中哪种物质发挥主效作用还有待探究。

IFN-γ能抑制内源性肿瘤坏死因子的产生,并参与细胞免疫,是重要的免疫调节因子[21]。IL-6、IL-1β是重要的炎症指标,当机体发生炎症时,炎性小体等会产生IL-6和IL-1β,导致血液中两者的浓度升高[22]。在本试验中,与对照组相比,添加50、100和150 g/(d·头)青蒿提取物均显著降低了奶牛血浆中IL-1β的浓度,同时血浆中IFN-γ的浓度有提高的趋势,说明在饲粮中添加青蒿提取物对奶牛的免疫性能具有提高作用,这也与血浆中免疫球蛋白浓度的变化相对应。帅学宏等[23]研究证明,青蒿提取物中的多糖可以促进淋巴细胞分泌IFN-γ,具有较好的抗炎作用。Park等[6]发现,青蒿提取物可以降低血液中IL-1β的浓度。上述两者的试验结果与本试验结果一致,都证明青蒿提取物具有提高动物机体免疫的作用,但本试验中奶牛血浆中IL-6浓度在青蒿提取物添加水平为50和100 g/(d·头)时相对于对照组降低但不显著,在添加水平为150 g/(d·头)时较对照组显著升高,且血浆中IgG的浓度变化异常,这说明添加150 g/(d·头)的青蒿提取物可能会引起奶牛产生应激,使体内部分生理指标处于炎症状态,对奶牛健康产生不利影响。由于免疫指标相互联系复杂交错,青蒿提取物在免疫方面发挥作用的具体机制还尚未明确。

3.3 青蒿提取物对奶牛血浆抗氧化指标的影响

氧化应激是影响奶牛健康的重要因素,当奶牛受到外界某些刺激时,其体内会产生大量自由基引起氧化应激,降低机体免疫力,影响生产性能[24]。研究发现,动物能通过酶机制和非酶机制消除体内的自由基,其中抗氧化酶主要包括SOD、CAT、GSH-Px等[25]。自由基作用于脂质后发生过氧化反应产生MAD,其能引起蛋白质、核酸等发生交联聚合,具有细胞毒性,能造成细胞膜脂质过氧化,破坏细胞膜结构完整性[26]。有研究表明青蒿提取物中的多糖、精油类物质可以通过清除自由基发挥抗氧化作用[27-28]。Iqbal等[29]发现青蒿提取物中的多酚、黄酮等物质在抗氧化过程中发挥着主要作用。张滔滔等[30]在肉鸡饲粮中添加青蒿提取物后,发现青蒿提取物可以提高肉鸡血液中GSH-Px与SOD的活性,降低MDA的浓度,这与本研究结果一致。青蒿提取物通过改善奶牛血液中GSH-Px与SOD的活性,清除自由基,降低MDA的浓度,从而提高奶牛的机体免疫力,其发挥作用的物质可能是青蒿中的多糖、多酚和黄酮类等物质。

4 结论

青蒿提取物能提高奶牛的产奶量、乳糖率、4%乳脂校正乳产量以及血浆中IgM和IFN-γ的浓度,降低血浆中MDA、IL-1β的浓度,能增强奶牛机体的免疫能力及抗氧化能力,且在100 g/(d·头)的添加水平下效果最佳。

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