复方中草药对中华鳖肠道微生物菌群及非特异性免疫功能的影响

包宇航; 章新宇; 尹尚军; 张海琪; 徐洁皓

doi:10.12131/20230069

复方中草药对中华鳖肠道微生物菌群及非特异性免疫功能的影响

doi: 10.12131/20230069

1.
浙江万里学院生物与环境学院，浙江宁波 315000
2.
浙江省淡水研究所/农业农村部淡水渔业健康养殖重点实验室，浙江湖州 313000

基金项目: 浙江省“三农六方”科技协作计划 (2021SNLF026)；浙江省水产新品种选育项目 (2021C02069-8-3)；宁波市重点技术研发项目(2021Z009)；浙江省“生物工程”一流学科 (A类) 学生创新计划项目 (CX2021017)

详细信息

作者简介:
包宇航 (1996—)，女，硕士研究生，研究方向为中华鳖免疫防抗。E-mail: 1620916838@qq.com

通讯作者:
徐洁皓 (1988—)，女，讲师，博士，研究方向为中华鳖免疫防抗。E-mail: xujiehao@zwu.edu.com

中图分类号: S 947.1
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- 文章访问数: 150
- HTML全文浏览量: 24
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- 被引次数: 0
出版历程
- 收稿日期: 2023-04-06
- 修回日期: 2023-05-15
- 录用日期: 2023-06-06
- 网络出版日期: 2023-06-09
- 刊出日期: 2023-10-05

Effects of Chinese herbal compound on intestinal microbiota and non-specific immune function of Pelodiscus sinensis

1.
School of Biology and Environment, Zhejiang Wanli University, Ningbo 315000, China
2.
Zhejiang Freshwater Research Institute/Key Laboratory of Freshwater Fishery Healthy Farming, Ministry of Agriculture and Rural Affairs, Huzhou 313000, China

摘要

摘要: 研制一种可提高中华鳖 (Pelodiscus sinensis) 免疫力的复方中草药，探究其对肠道微生物菌群的调节作用，以为中华鳖病害防治提供参考。将平均体质量为 (44.59±9.23) g的100只中华鳖分为两组，对照组投喂基础饲料，实验组投喂含2% (质量分数) 复方中草药的饲料，连续投喂42 d。结果显示，两组优势菌门相同，但其所占比例存在差异：实验组中华鳖肠道中软壁菌门、浮霉菌门和乳杆菌属 (Lactobacillus) 等有益菌群丰度分别是对照组的32、2.25和4.12倍。利用10⁷ CFU·mL⁻¹浓度的嗜水气单胞菌 (Aeromonas hydrophila) 攻毒，测定中华鳖血清内的溶菌酶 (LZM)、总超氧化物歧化酶 (T-SOD)、碱性磷酸酶 (AKP) 和酸性磷酸酶 (ACP) 的活性。结果显示，对照组和实验组的T-SOD、LZM、ACP及AKP活性均在第72小时达到峰值，且实验组中华鳖血清中的4种酶在正常状态下及攻毒后的活性几乎均显著高于对照组，除LZM在攻毒12 h后两组差异不显著外。研究表明，复方中草药可有效优化中华鳖肠道微生物菌群结构，具有一定的免疫增强效果，应用前景良好。
- 中华鳖 /
- 中草药 /
- 非特异性免疫 /
- 肠道菌群
Abstract: We developed a Chinese herbal compound to improve the immunity of Chinese soft-shelled turtle (Pelodiscus sinensis) and invetigated its effect on the regulation of intestinal microflora, so as to provide references for its disease control. In this study, we divided 100 individuals of P. sinensis with an average body mass of (45.53±3.89) g into two groups. The control group (CG) was fed with a basal diet and the experimental group (CPG) was fed with a diet supplemented with 2% of compound herbs for 42 d. The results show that the dominant phylum was the same in both groups, but the proportion of each dominant bacterial phylum differed: the beneficial flora of Tenericutes, Planctomycetes and Lactobacillus in the intestine of the CPG group was 32, 2.25 and 4.12 times higher than that of the CG group, respectively. After being challenged by 10⁷ CFU·mL⁻¹ of Aeromonas hydrophila, the activity of lysozyme (LZM), total superoxide dismutase (T-SOD), alkaline phosphatase (AKP) and acid phosphatase (ACP) in serum of P. sinensis were determined. The results show that the activities of T-SOD, LZM, ACP and AKP in the CG and CPG groups reached the peak values at 72^nd hour, and activities of the four enzymes in the serum of CPG group were significantly higher than those in CG group under the normal conditions and after the challenge , except for LZM, which was not significantly different when the two groups were challenged for 12 h. In conclusion, this Chinese herbal compound can effectively optimize the intestinal microflora of P. sinensis and has certain immune enhancing effects, which shows a application prospect.
- Pelodiscus sinensis /
- Chinese herbal compound /
- Non-specific immune function /
- Intestinal flora

HTML全文

图 1 Alpha 多样性指数 box 图

注：对照组投喂基础饲料；实验组投喂添加2%的复方中草药的饲粮。后图同此。

Figure 1. Alpha diversity index box chart

Note: CG. Feeding basal diet; CPG. Feeding diet with 2% of Chinese herbal compound. The same case in following figures.

下载: 全尺寸图片幻灯片

图 2 对照组和实验组肠道菌群组成丰度差异 (门水平)

Figure 2. Relative abundance of intestinal microflora composition of CG and CPG at genus level

下载: 全尺寸图片幻灯片

图 3 对照组和实验组肠道菌群组成丰度差异 (属水平)

Figure 3. Relative abundance of intestinal microflora composition of CG and CPG at genus level

下载: 全尺寸图片幻灯片

图 4 对照组和实验组肠道菌群 LEfSe 差异分析图

注：无显著差异的物种统一着色为黄色，差异物种跟随组进行着色，红色节点表示在红色组别中起到重要作用的微生物类群，绿色节点表示在绿色组别中起到重要作用的微生物类群；圆圈大小与微生物相对丰度成正比例关系。

Figure 4. LEfSe variance analysis of intestinal microflora of CG and CPG

Note: Species without significant differences are uniformly colored in yellow, and differential species are colored following the group; red nodes represent microbial taxa that play an important role in the red group, and green nodes represent microbial taxa that play an important role in the green group; the size of the circle is proportional to the relative abundance of microorganisms.

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图 5 对照组和实验组肠道菌群功能预测 STAMP 差异分析

注：左图所示为不同微生物功能预测在两组样本中的丰度比例，右图为95%置信度区间内微生物功能预测丰度的差异比例；左边纵坐标轴为不同的微生物功能预测分类。

Figure 5. STAMP variance analysis of functional prediction of intestinal microflora of CG and CPG

Note: The left panel shows the proportion of abundance of different microbial functional predictions in the two groups of samples, and the right panel shows the proportion of differences in the abundance of microbial functional predictions within the 95% confidence interval; the left vertical axis shows different microbial functional prediction classifications.

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图 6 复方中草药对嗜水气单胞菌刺激前后中华鳖血清中总超氧化物歧化酶、溶菌酶、酸性磷酸酶和碱性磷酸酶活性的影响

注：误差线用标准误值 (n=6) 表示；*. P<0.05，**. P<0.01，***. P<0.001，ns表示无显著性差异。

Figure 6. Effect of Chinese herbal compound on T-SOD, LZM, ACP and AKP activity in serum of P. sinensis before and after A. hydrophila challenge

Note: The error bar is expressed by the SEM value (n=6); *. P<0.05, **. P<0.01, ***. P<0.001; ns. No significant differences.

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