YIC-IR
菊芋全粉发酵饲料在仿刺参养殖中的应用效果研究
Alternative TitleApplication Effects of Fermented Feed with Jerusalem Artichoke Powder in Apostichopus japonicus Culture
杨燕云
Subtype硕士
Thesis Advisor衣悦涛
2022-05-12
Training institution中国科学院烟台海岸带研究所
Degree Grantor中国科学院烟台海岸带研究所
Keyword菊芋全粉 益生元 仿刺参 肠道微生态 生长性能
Abstract随着养殖密度的增加,仿刺参养殖面临着细菌性传染病的困扰。为了寻找绿色安全的手段改善仿刺参肠道生态健康,促进仿刺参健康养殖,本论文将富含菊糖的菊芋全粉作为益生元成分加入微生物固态发酵饲料中,制备成菊芋全粉发酵饲料。并将其应用到仿刺参人工养殖中,探究菊芋全粉和发酵饲料对仿刺参肠道微生物、肠道短链脂肪酸含量、肠道形态结构、消化酶、非特异性免疫、生长以及体壁营养组分的影响,同时考察二者在仿刺参养殖中的协同效果。主要研究内容及结论如下: 1. 将酿酒酵母(Saccharomyces cerevisiae)、枯草芽孢杆菌(Bacillus subtilis)和保加利亚乳杆菌(Lactobacillus Bulgaricus)分别扩大培养后按比例混合得到复合菌剂(8.75×109 CFU/mL),对仿刺参基础饲料粗粉进行固态发酵培养制得发酵饲料,并对发酵前后营养成分进行测定。结果显示发酵后粗蛋白含量提高,粗纤维含量下降,粗脂肪含量无明显变化。发酵后的饲料营养成分配比优于基础饲料,有利于仿刺参的生长。 2. 将仿刺参随机分为四组,分别为对照组(CT组,基础饲料)、发酵饲料组(FF组)、菊芋全粉基础饲料组(IBF组,1.5%菊芋全粉)和菊芋全粉发酵饲料组(IFF组,1.5%菊芋全粉),每组6个平行,每个平行10只仿刺参,进行为期60 d的仿刺参养殖试验。养殖试验结束后对仿刺参肠道菌群多样性和物种丰度、肠道内容物短链脂肪酸(SCFAs)含量、肠道组织形态、消化酶活性、免疫力、生长性能和体壁营养成分进行测定。 (1)菊芋全粉发酵饲料对肠道微生物和SCFAs含量的影响:与对照组相比,实验组仿刺参肠道微生物的多样性和丰富度均显著提高(P<0.05),IFF组肠道微生物与对照组同源性更强。在门水平上,FF组和IFF组变形菌门(Proteobacteria)丰度显著降低、厚壁菌门(Firmicutes)丰度显著提高(P<0.05),IBF组变形菌门(Proteobacteria)和疣微菌门(Verrucomicrobiota)丰度显著降低(P<0.05)。在目水平上,IBF组和IFF组红杆菌(Rhodobacterales)丰度显著降低(P<0.05),黄杆菌(Flavobacteriales)丰度增加,IFF组葡萄球菌(Staphylococcales)和芽孢杆菌(Bacillales)丰度显著提高(P<0.05)。另外,相较于对照组,实验组总SCFAs含量均有所提高。IBF组乙酸含量显著提高(P<0.05),实验组丁酸含量均有提高,但无显著差异(P>0.05)。 (2)菊芋全粉发酵饲料对肠道组织形态和消化酶活力的影响:与对照组相比,FF组仿刺参肠道绒毛高度和微绒毛数量增加;IBF组前肠绒毛宽度降低,中肠微绒毛数量增加,肠道肌层厚度提高;IFF组肠道绒毛高度及数量增加,各实验组肠道绒毛发育和肌层厚度均有不同程度的改善。另外,FF组蛋白酶、淀粉酶、脂肪酶和纤维素酶活性显著提高(P<0.05),IBF组消化酶活性提高效果不明显(P>0.05),IFF组蛋白酶、纤维素酶和脂肪酶活性显著提高(P<0.05)。 (3)菊芋全粉发酵饲料对免疫力的影响以及免疫酶活与肠道菌群相关性分析:与对照组相比,FF组和IFF组仿刺参体腔液中酚氧化酶(PO)活性显著提高,但一氧化氮合酶(NOS)活性显著降低(P<0.05);IBF组和IFF组体腔细胞总数(TCC)和吞噬活性显著提高(P<0.05);IFF组酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性也显著提高(P<0.05);各实验组饲料对超氧化物歧化酶(SOD)活性影响不显著(P>0.05)。通过免疫酶和抗氧化酶与肠道菌群相关性分析发现,NOS活性与假鲁杰氏菌属(Pseudoruegeria)、芽孢杆菌属(Bacillaceae)、海岸杆菌属(Actibacter)、葡萄球菌属(Staphylococcus)丰度呈显著负相关,与鲁杰氏菌属(Ruegeria)、洛克氏菌属(Halocynthiibacter)和Vermiphilaceae呈显著正相关(P<0.05)。SOD和PO活性分别与热带杆菌属(Tropicibacter)和假鲁杰氏菌属(Pseudoruegeria)丰度呈显著正相关,但ACP、AKP活性与洛克氏菌属(Halocynthiibacter)呈显著负相关(P<0.05)。 (4)菊芋全粉发酵饲料对生长性能和体壁营养组分的影响:在生长性能方面,与对照组相比,实验组饲料均显著提高了养殖仿刺参的增重率和特定生长率,降低了饲料系数(P<0.05),提高了饲料利用率。在大规模养殖试验中,菊芋全粉发酵饲料将增重率显著提高(P<0.05)。在体壁营养成分上,实验组饲料对水分和灰分无明显影响(P>0.05),对粗蛋白水平有一定的提高作用,并显著降低了粗脂肪含量(P<0.05)。 本研究结果表明,菊芋全粉发酵饲料能够改善仿刺参肠道菌群结构,促进芽孢杆菌等益生菌的增殖并提高肠道内短链脂肪酸含量。对促进肠道组织形态结构的发育和消化酶活性的提高有明显效果,还能提高免疫力和生长性能以及改善仿刺参品质,证明菊芋全粉和发酵饲料能够发挥协同效果。本研究为菊芋全粉发酵饲料在仿刺参养殖中的应用提供了数据支撑。
Other AbstractWith the increase of breeding density, the cultivation of sea cucumber faces the trouble of bacterial infectious diseases. It is necessary to search for a green and safe way to improve the intestinal ecological health and promote the healthy cultivation of sea cucumber Apostichopus japonicus. Therefore, in this dissertation, the Jerusalem artichoke powder (JAP) was added to the microbial solid-state fermentation feed. JAP is rich in inulin and can be used as a prebiotic ingredient. In this study, the fermented feed with JAP was applied to the artificial culture of Apostichopus japonicus. The purpose of this test is to investigate the effects of Jerusalem artichoke powder and fermented feed on the gut microbiota, intestinal short-chain fatty acid content, intestinal morphology and structure, digestive enzymes, non-specific immunity, growth and body nutritional composition of Apostichopus japonicus, and to study the synergistic effects of them. The main research contents and conclusions of the dissertation are as follows: 1. The Saccharomyces cerevisiae, Bacillus subtilis, and Lactobacillus Bulgaricus were cultured respectively and mixed in proportion to obtain compound probiotics (8.75×109 CFU/mL). The crude powder of basal diet was cultured by solid-state fermentation with probiotics, and the nutritional components were measured before and after fermentation. The results showed that the crude protein content was increased, the crude fiber content was reduced, and the crude fat content did not change significantly. The nutrient composition ratio of fermented feed was better than that of basal diet, which was beneficial to the growth of Apostichopus japonicus. 2. In this work, 240 sea cucumbers were randomly divided into 4 groups, each group had 6 replicates, and each replicate had 10 sea cucumbers. The groups were as follows: Control group (CT group, basal diet), Fermented Feed group (FF group), JAP Basic Feed group (IBF group, 1.5% JAP), and JAP Fermented Feed group (IFF group, 1.5% JAP). After feeding for 60 days, the diversity of intestinal flora, the content of short-chain fatty acids (SCFAs) in intestinal contents, the morphology of intestinal tissues, the activities of digestive enzymes, immunity, growth performance, and body wall nutritional composition of Apostichopus japonicus were measured. (1) The effect of fermented feed with JAP on the gut microbiota and intestinal short-chain fatty acid content: Compared with the control group, the diversity and richness of intestinal microflora in the experimental groups were significantly increased (P<0.05), and the intestinal microorganisms of the IFF group with the control group were more homologous. At the phylum level, compared with the control group, the abundance of Proteobacteria was reduced significantly and the abundance of Firmicutes was increased significantly in the FF group and IFF group (P<0.05). Meanwhile, the abundance of Proteobacteria and Verrucomicrobiota was significantly reduced in the IBF group (P<0.05). At the order level, the abundance of Rhodobacterales was significantly reduced in the IBF group and IFF group (P<0.05) but the abundance of Flavobacteriales was increased, and the abundance of Staphylococcales and Bacillales was significantly increased in the IFF group (P<0.05). In addition, compared with the control group, the total SCFAs contents were increased in all the experimental groups. The content of acetic acid was significantly increased in the IBF group (P<0.05), the contents of butyric acid were increased in all the experimental groups, but there was no significant difference (P>0.05). (2) The effect of fermented feed with JAP on the morphology of intestinal tissues and the activities of digestive enzymes: Compared with the control group, the intestinal villi height and microvilli number in the FF group were increased. In the IBF group, the width of foregut villi was reduced, the number of midgut microvilli was increased, and the thickness of intestinal muscle layer was increased. The height and number of intestinal villi were increased in the IFF group. The feed of experimental groups all contributed to varying degrees to promoting the development of intestinal villi and the improvement of muscle layer thickness. In addition, the activities of protease, amylase, lipase, and cellulase in the FF group were significantly increased (P<0.05). The feed of IBF group had no significant effect on the improvement of digestive enzyme activity (P>0.05). In the IFF group, the activities of protease, cellulase, and lipase were significantly increased (P<0.05). (3) The effect of fermented feed with JAP on immunity and the correlation analysis between immune enzyme activities and intestinal flora: Compared with the control group, the feed of FF and IFF groups significantly increased the activity of phenol oxidase (PO), but significantly reduced the activity of nitric oxide synthase (NOS) (P<0.05). The total number of coelomic cells (TCC) and phagocytic activity were significantly increased in the IBF and IFF groups (P<0.05). The activities of acid phosphatase (ACP) and alkaline phosphatase (AKP) in the IFF group were also significantly increased (P<0.05). The feed of experimental groups had no significant effect on superoxide dismutase (SOD) activity (P>0.05). The correlation analysis between immune enzymes activities and antioxidant enzymes activities with intestinal flora showed that NOS activity was negatively correlated with the abundance of Pseudoruegeria, Bacillaceae, Actibacter, and Staphylococcus and positively correlated with the abundance of Ruegeria, Halocynthiibacter, and Vermiphilaceae (P<0.05). The activities of SOD and PO were positively correlated with the abundance of Tropicibacter and Pseudoruegeria, respectively (P<0.05). However, ACP and AKP activities were negatively correlated with Halocynthiibacter (P<0.05). (4) The effect of fermented feed with JAP on growth performance and body wall nutritional composition: In terms of growth performance, compared with the control group, the feed of experimental groups all significantly increased the weight gain rate (WGR) and special growth rate (SGR), reduced the feed coefficient (FC) (P<0.05), and improved the feed utilization rate of Apostichopus japonicus. In the large-scale feeding trial, the fermented feed with JAP increased WGR significantly (P<0.05). In terms of body nutritional composition, the feed of experimental groups had no significant effect on moisture and ash content (P>0.05) but increased the level of crude protein to a certain extent, and significantly reduced the content of crude lipid (P<0.05). The results of this study showed that the fermented feed with JAP can improve the intestinal microflora, promote the proliferation of probiotics such as Bacillus and increase the intestinal short-chain fatty acid content. At the same time, it can promote the development of intestinal morphology and digestive enzyme activities, improve immunity and growth performance and improve the quality of Apostichopus japonicus. Moreover, the combination uses of 1.5% JAP and fermented feed showed better synergistic effects. This study provides a data support for the application of the fermented feed with JAP in the cultivation of sea cucumbers.
Pages113
Language中文
Document Type学位论文
Identifierhttp://ir.yic.ac.cn/handle/133337/30832
Collection中国科学院烟台海岸带研究所
Recommended Citation
GB/T 7714
杨燕云. 菊芋全粉发酵饲料在仿刺参养殖中的应用效果研究[D]. 中国科学院烟台海岸带研究所,2022.
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