【作者】 刘旭；

【导师】 罗燕； 程建国；

【作者基本信息】 四川农业大学 ， 兽医硕士（专业学位）， 2020， 硕士

【摘要】 由于滥捕乱杀,林麝濒临灭绝,已被列为我国一级保护动物。人工圈养虽然取得了一定的成功,但健康问题一直制约着人工养麝的发展。菌群失调是引起林麝肠道疾病的重要因素,饲喂微生态制剂有利于宿主肠道菌群平衡的改善和恢复。本研究通过测定对照组(4头)、试验组A(4头)和试验组B(4头)的平均日增重(Average Daily Gain,ADG)和血清中几种免疫相关因子含量分析林麝复合微生态制剂对育成林麝体重及免疫的影响;并利用16S rRNA高通量测序探究林麝复合微生态制剂对育成林麝粪便菌群的影响;此外,采用活菌计数法测定粪便中乳酸菌的数量。主要结果如下:1.林麝复合微生态制剂对育成林麝体重、免疫及粪便中乳酸菌数量的影响:饲喂林麝复合微生态制剂后,对照组与试验组育成林麝平均日采食量无显著差异,试验组ADG显著高于对照组(P<0.05),料重比显著低于对照组(P<0.05)。试验组B血清中Ig A和Ig G的增长量显著高于对照组(P<0.05),TNF-α、IFN-γ、Ig M和IL-2含量没有显著变化(P>0.05)。试验组B粪便中的乳酸菌数量显著高于对照组(P<0.05),试验组A与对照组粪便中的乳酸菌数量无显著性差异(P>0.05)。2.林麝复合微生态制剂对育成林麝肠道菌群的影响:粪便样品共检出的细菌有21门,37纲,63目,132科,256属。饲喂林麝复合微生态制剂后,对照组和试验组的共有OTU数量增加;试验组B的Chao1、Shannon和Simpson指数均升高且具有统计学意义(P<0.05),其中Shannon指数差异极显著(P<0.01),ACE指数无明显变化;对照组和试验组A的Chao1、ACE、Shannon和Simpson指数均无明显变化(P>0.05),且对照组的Chao1和ACE指数均呈下降趋势。在主成分分析图中,试验组A和试验组B样出现聚类现象,对照组仍然较分散。饲喂林麝复合微生态制剂后试验组的肠道菌群在门水平和属水平上出现较大变化。在门水平上,变形杆菌(Proteobacteria)的相对丰度均显著降低(P<0.05),厚壁菌门(Firmicutes)和疣微菌门(Verrucomicrobia)相对丰度均显著增加(P<0.05);在属水平上,假单胞菌(Pseudomonas)相对丰度均显著降低(P<0.05),瘤胃球菌(Ruminococcus)、阿克曼菌属(Akkermansia)、颤螺旋菌属(Oscillospira)、粪球菌属(Coprococcus)和梭菌属(Clostridium)相对丰度显著升高(P<0.05)。综上所述,林麝复合微生态制剂可以促进育成林麝体重的增长,提高育成林麝机体免疫球蛋白含量,平衡育成林麝肠道菌群,增加肠道菌群多样性和丰富度,促进肠道有益菌群的生长。为复合微生态制剂在林麝人工养殖中的广泛应用奠定重要基础。更多还原

【Abstract】 Forest musk deer(FMD,Moschus berezovskii)are endangered due to indiscriminate hunting and killing and FMD were listed as the first-class protected animal.Some success has been achieved in captivity,but the health problems of FMD restrict the development of captive FMD.Dysbacteriosis is an important factor causing intestinal diseases of FMD.Probiotics are intended to provide health benefits when consumed,generally by improving or restoring the gut flora.This study was conducted with the aim of analyzing the effects of forest musk deer compound probiotics(FMDPs)on weight and immunity in growing FMD by measuring average daily weight gain(ADG)and immune-related factors and by using high-throughput16 S rRNA sequencing to investigate differences in the gut microbiota among the control group(4 samples),treatment group A(4 samples)and treatment group B(4 samples).In addition,the number of lactic acid bacteria in feces was measured by the viable plate count method.1.Effects of compound probiotics on the weight,immunity performance and the number of lactic acid bacteria in feces of forest musk deer: After FMDP feeding,there were no significant differences in the average daily feed intake of three groups and the average daily weight gain(ADG)of the treatment groups was significantly higher than that of the control group(P<0.05).The feed/gain(F/G)ratio was significantly lower than that of the control group(P<0.05).The increase in Ig A and Ig G levels in treatment group B was significantly higher than that in the control group(P<0.05),and there was no significant change in TNF-α,IFN-γ,Ig M and IL-2 levels(P>0.05).2.Effects of compound probiotics on the gut microbiota of forest musk deer: The bacteria that could be tested were classified into 21 phyla,37 classes,63 orders,132 families,and 256 genera.After FMDP feeding,the number of components shared by the control group and treatment group A increased,the Chao1,Shannon and Simpson indices of treatment group B increased after FMDP feeding,and the difference was statistically significant(P<0.05).The Shannon index showed a marked difference(P<0.01),but there was no significant change in the ACE index.There was no significant change in the Chao1,ACE,Shannon and Simpson indices of the control group and treatment group A after FMDP feeding(P>0.05),and the Chao1 and ACE indices of the control group both showed a downward trend.In PCo A,samples of treatment group A and treatment group B exhibited obvious clustering after FMDP feeding,and samples of control group were still scattered.The gut microbiota of the treatment groups were significantly changed at phylum level and genus level after FMDP feeding.At the phylum level the relative abundance of Proteobacteria was significantly lower than on day 0(P<0.05),the relative abundances of Firmicutes and Verrucomicrobia were significantly higher than those on day 0(P<0.05).At the genus level,the relative abundance of Pseudomonas was significantly decreased(P<0.05),and the relative abundances of Ruminococcus,Akkermansia,Oscillospira,Coprococcus and Clostridium were significantly increased(P<0.05).In summary,the results indicated that FMDPs could promote the growth of growing FMD,improve immunity,balance the role of intestinal microbes,increase the diversity and richness of gut microbiota and promote the growth of beneficial bacteria in the intestinal tract.It lays an important foundation for the extensive application of FMDP in artificial breeding of forest musk deer更多还原