一株蛋鸡肠道枯草芽孢杆菌的分离鉴定与益生菌对鸡的应用效果

发布时间：2018-06-16 23:36

本文选题：益生素 + 16s　；参考：《山东农业大学》2015年硕士论文

【摘要】：我国是个畜牧养殖大国,在养殖过程中畜禽细菌性疾病基本上都是用抗生素治疗,随着抗生素的大量使用,一系列的问题也随之产生:抗生素在解决病原菌、抑制杀灭病原菌的同时,也使病原微生物产生了对抗生素的耐药性,据农业部报道,目前,几乎没有一种抗生素不存在耐药现象,病原菌耐药菌株的形成已经成了一个全球性热烈讨论及难以解决的问题,耐药性的形成不仅使抗生素的疗效降低,治疗用药物剂量增大、疾病的复发率升高、治疗周期延长,而会引起疾病并发症,导致动物死亡率升高等。寻找一种抗生素的替代品成为了比较迫切的问题,其中微生态制剂最先被推到了最前沿,那么,微生态是否能起到人们期待的效果呢?因此笔者从饲喂益生菌后的蛋鸡肠道分离了有益菌---枯草芽孢杆菌,还研究了饲喂益生菌后对肉鸡和蛋鸡的生产性能、粪便排泄指标以及其他各指标的影响并进行差异显著性分析。本实验通过对蛋鸡肠道有益菌的分离、染色镜检、生化试验、16s rRNA测序并通过NCBI的Blast检索系统进行序列同源性分析,然后与从GenBank数据库中获得的枯草芽孢杆菌的16s rRNA序列进行多序列匹配,采用邻接法构建系统进化树,还对肉鸡的料肉比,蛋鸡的料蛋比、蛋重、抗体效价、血细胞成分以及粪便排泄指标中的粗蛋白、灰分、钙、磷等指标进行测定,试验中主要采用凯式定氮法、高锰酸钾法等。试验结果表明:蛋鸡肠道分离的有益菌菌株经16s rRNA基因测序序列与枯草芽孢杆菌D53株的同源性为99%以上,表明有益菌能在鸡肠道存在并分离,从而发挥作用。蛋鸡试验结果:使用益生菌后,试验组2组和3组的蛋重与对照组的差异显著(P0.05),试验组的蛋重比对照组的蛋重大0.96-2.5g;在次(破、软)蛋率上,实验2组和3组与对照组相比差异显著(P0.05),试验组比对照组降低23.9%-28.4%;在料蛋比上,试验1组和3组与对照组相比差异显著(P0.05),试验组比对照组降低4.65%,蛋壳厚度和蛋长宽比差异不显著;粪便排泄指标成分测定结果显示:试验组灰分、Ca的含量与对照组差异显著(P0.05),试验组的灰分百分含量比对照组高了4.4个百分点;试验组的Ca百分含量小于对照组,比对照组低了1个百分点;磷的含量差异不显著(P0.05),但从数据上看,试验组的百分含量小于对照组;试验组的ND、H5、H9的抗体效价比对照组效价高;实验组的淋巴细胞数与对照组差异显著(P0.05),白细胞总数、淋巴细胞数、中间细胞以及粒细胞数与对照组差异不显著,但数值上实验组比对照组数量大。从以上试验结果表明:蛋鸡使用益生素后能增加蛋的质量和品质,降低料蛋比,降低次(破、软)蛋率;能提高饲料吸收率;提高鸡体的抗病力、免疫保护力。在肉鸡实验中,使用益生菌后能降低肉鸡的料肉比,能保持肠道菌群平衡,增加饲料中蛋白、钙、磷的吸收,减少营养物质的排放,减少生产成本,提高养殖业的经济效益。
[Abstract]:China is a big animal breeding country. In the process of breeding, the bacterial diseases of livestock and poultry are basically treated with antibiotics. With the use of antibiotics, a series of problems are also produced: antibiotics are used to solve pathogenic bacteria, inhibit the killing of pathogenic bacteria, and also cause the pathogenic microorganism to produce antibiotic resistance. According to the Ministry of agriculture, the Ministry of Agriculture reported that At present, there is almost no antibiotic resistance, and the formation of drug-resistant strains of pathogens has become a global and difficult problem. The formation of drug resistance not only reduces the efficacy of antibiotics, increases the dosage of the drug, increases the recurrence rate of the disease, and prolongs the treatment cycle, which will cause the disease and cause the disease. It is an urgent problem to find an alternative to an antibiotic, and the microecology is the first to be pushed to the forefront. Then, can microecology be expected to be expected? So I isolated the beneficial bacteria, Bacillus subtilis, from the gut of the egg chicken fed by the probiotics. The effect of probiotics on the production performance, fecal excretion index and other indexes after feeding probiotics was studied and the difference was analyzed. This experiment was carried out by sequencing of the beneficial bacteria in the intestinal tract, staining microscopy, biochemical test, 16S rRNA sequencing and the sequence homology analysis through the Blast retrieval system of NCBI, and then, The 16S rRNA sequence of Bacillus subtilis obtained from the GenBank database was matched by multiple sequences, and the phylogenetic tree was constructed by adjacency method. The indexes of meat and meat ratio, egg weight, egg weight, antibody titer, blood cell composition and fecal excretion index were measured. The results showed that the homology of the beneficial bacteria isolated from the gut of the laying hens was more than 99% by the sequence of 16S rRNA gene sequencing and D53 strain of Bacillus subtilis D53. The results showed that the beneficial bacteria could exist and separate in the chicken gut. The results of the egg laying test: after the use of probiotics, 2 groups and 3 groups of the test group were used. The egg weight of the test group was significantly different from the control group (P0.05). The egg weight of the test group was significantly higher than that of the control group. On the second (broken, soft) egg rate, the 2 and 3 groups were significantly different from the control group (P0.05), and the experimental group was lower than the control group (23.9%-28.4%); the 1 groups and 3 groups were significantly different from the control group (P0.05), and the test group was compared with the control group (P0.05). The control group decreased by 4.65%, the thickness of eggshell and the egg length width ratio of the control group was not significant. The results of the fecal excretion indexes showed that the ash content of the test group was significantly different from the control group (P0.05), the ash content of the test group was 4.4 percentage points higher than that of the control group, and the Ca percent content of the test group was less than the control group, which was 1 100% lower than the control group. The difference of phosphorus content was not significant (P0.05), but from the data, the percentage of test group was less than that of the control group; the antibody titer of ND, H5 and H9 in the experimental group was higher than the control group; the number of lymphocyte in the experimental group was significantly different from the control group (P0.05), the total number of leukocytes, the number of lymphocytes, the intermediate cells and the number of granulocytes were different from the control group. It is not significant, but the number of the experimental group is larger than the control group. The results from the above test show that the use of probiotics can increase the quality and quality of eggs, reduce the ratio of egg to eggs, reduce the rate of secondary (broken, soft) eggs, improve the feed absorption rate, improve the resistance and immunity of the chicken body. In the broiler experiment, the use of probiotics can reduce the broiler. The ratio of meat and meat can maintain the balance of intestinal flora, increase the absorption of protein, calcium and phosphorus in the feed, reduce the emission of nutrients, reduce the production cost, and improve the economic benefit of the breeding industry.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831.5

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