不同氮源对猪肠道微生物多样性及主要产蛋白酶菌株的影响
发布时间：2018-10-31 19:21
【摘要】：为了探究低蛋白日粮中,不同蛋白源对猪肠道微生物的影响,本研究以棉籽粕、鱼粉和玉米胚芽粕等作为主要蛋白来源,考察了低蛋白日粮对仔猪和育肥猪肠道微生物多样性和主要产蛋白酶菌株的影响。本研究共包括三个试验。试验一考察低蛋白日粮不同氮源对断奶仔猪小肠微生物的影响。将48头断奶仔猪(14.62±1.46 kg),随机分为4组,每组3个重复,每个重复4头仔猪,仔猪采用单笼饲养。试验处理为对照组(SBM组)、棉籽粕(CSM)、豆粕-棉籽粕(SC)和鱼粉(FM)组,其中CSM组和SC组日粮中棉籽粕分别替代对照组豆粕提供粗蛋白的100%和50%。日粮以净能体系配制,补充平衡各组必需氨基酸。饲养试验结束时,每个重复随机选取1头仔猪进行屠宰、取样利用Illumina Miseq测序技术测定十二指肠、空肠前段、中段、后段和回肠微生物多样性。研究结果表明,不同蛋白源组仔猪小肠微生物多样性和主要微生物组成差异均较大。其中,Firmicutes、Proteobacteria、Bacteroidetes和Acidobacteria在各组丰度均较高,添加棉籽粕两组中有益菌Lactobacillus和Clostridium丰度较其他组更高,而FM组中Escherichia/Shigella丰度较高。由此得出,不同蛋白源对仔猪小肠微生物多样性有较大影响,棉籽粕替代豆粕日粮提高了仔猪小肠有益菌比例。试验二研究了低蛋白日粮不同氮源对育肥猪生长性能和肠道微生物的影响。选取36头杜长大育肥阉公猪(58.65±3.71 kg)随机分在4组中,每个处理3个重复,每个重复3头猪。处理组为对照组(SBM15)组和三个低蛋白(11%CP)日粮组,其中豆粕(SBM11)、棉籽粕和玉米胚芽粕50%等氮替代豆粕(SCCM)、棉籽粕和玉米胚芽粕100%等氮替代豆粕(CCM)分别作为三种低蛋白日粮中主要蛋白源。日粮以净能体系配制,补充平衡各组必需氨基酸。经过28 d饲喂后,取回肠内容物和粪便作为肠道微生物分析样本,通过Illumina Miseq测序技术测定样本微生物多样性。结果显示,SBM11与CCM组生长性能显著低于SBM15组(P0.05),但SCCM组与SBM15组生长性能没有显著差异(P0.05)。对16SrRNA V3-V4区进行扩增测序后发现,SCCM组日粮显著增加了育肥猪肠道微生物多样性,并提高了有益菌(如Lactobacillus和Bifidobacterium)丰度。除此之外,Ruminococcus、Prevotellaceae和Bacteroides相对丰度在低蛋白日粮组中也有显著提升。因此,豆粕、棉籽粕和玉米胚芽粕作为育肥猪日粮主要蛋白源即使在低蛋白的情况下,可以调节肠道微生物,维持育肥猪的正常生长。试验三从健康仔猪小肠内筛选一株产蛋白酶菌2L6,经形态、生理生化和分子生物学鉴定确定该菌株为蜡样芽胞杆菌(Bacillus cereus)。分别在豆粕、棉籽粕、豆粕-棉籽粕和鱼粉发酵培养基中对其进行发酵培养,并在不同时间检测蛋白酶活性和底物蛋白消化率。结果表明,菌株2L6在不同发酵培养基中蛋白酶活性及底物蛋白消化率有较大差异。其中,豆粕-棉籽粕混合发酵培养基中蛋白酶活性在相对较短时间内即达到最大值21.31±0.09U/m L,而底物蛋白消化率也在36 h即达到最大值64.42%(P0.01)。因此,豆粕-棉籽粕混合蛋白源可较大提高仔猪小肠主要蛋白酶菌(Bacillus cereus strain 2L6)产蛋白酶活力及其对蛋白质利用能力。综上所述,低蛋白日粮中以棉籽粕、玉米胚芽粕等杂粕作为主要蛋白来源,可以改善仔猪和育肥猪的肠道微生物组成,增高有益菌比例,提高肠道中主要产蛋白酶菌株活性,可作为低蛋白日粮中豆粕的合理替代选择。
[Abstract]:In order to explore the effect of different protein sources on the intestinal microorganisms in pigs, the effects of the diets on the microbial diversity and the major protease-producing strains of piglets and fattening pigs were studied by using cottonseed meal, fish meal and corn germ meal as the main source of protein. This study consisted of three trials. Effects of different nitrogen sources on small intestinal microorganisms in weanling piglets. Forty-eight weanling piglets (14.62. 1.46 kg) were randomly divided into 4 groups, 3 replicates in each group, 4 piglets were repeated, and the piglets were fed with single cage. The test was treated as control group (SBM group), cottonseed meal (CSM), soybean meal-cottonseed meal (SC) and fish meal (FM) group, among which, the cottonseed meal in the CSM group and SC group was 100% and 50% of the crude protein in the control group respectively. Daily grain is formulated in net energy system to supplement essential amino acids in each group. At the end of the breeding trial, 1 piglets were randomly selected for slaughter and sampling was performed using the Illumina Miseq sequencing technique to determine the diversity of the duodenum, the anterior segment, the middle segment, the posterior segment, and the ileum. The results showed that the diversity of small intestine and the main microbial composition of piglets in different protein source groups were different. Among them, Firmicutes, Proteobacteria, Bacteroidites and Acidobacter were higher in each group, and the abundance of Lactobacillus and Clostridia was higher in the two groups of cottonseed meal, while the abundance of Escherrez/ Shigella in FM group was higher. The results showed that different protein sources had a great influence on the microbial diversity of small intestine of piglets. The effects of different nitrogen sources on the growth and intestinal microorganisms of fattening pigs were studied. Thirty-six (58. 65/ 3. 71 kg) pigs were randomly divided into 4 groups, 3 replicates per process and 3 pigs per replicate. The treatment group was a control group (SBM15) group and three diets (11% CP) diet group, in which 50% of soybean meal (SBM15), cottonseed meal and corn germ meal were replaced by nitrogen instead of soybean meal (SCCM). Cotton seed meal and corn germ meal (100%) were used as the main protein sources in three diets. Daily grain is formulated in net energy system to supplement essential amino acids in each group. After 28 days feeding, ileal contents and feces were taken as intestinal microbial analysis samples, and the microbial diversity of samples was determined by Illumina Miseq sequencing technique. The results showed that the growth performance of SBM15 and CCM was significantly lower than that of SBM15 group (P0.05), but there was no significant difference between SCCM and SBM15 group (P0.05). After the amplification and sequencing of the 16SrRNA V3-V4 region, it was found that the SCCM group day grain significantly increased the microbial diversity of the fattening pigs and increased the abundance of bacteria such as Lactobacillus and Bifidobacterium. In addition, the relative abundance of Rhomoccu, Prevoteellaceae and Bactereoides was also significantly improved in the diet group. Therefore, the soybean meal, the cottonseed meal and the corn germ meal can be used as the main protein source of the daily grain of the fattening pig, the intestinal microorganism can be adjusted, and the normal growth of the fattening pig can be maintained. A protease-producing strain 2L6 was screened from the small intestine of healthy piglets, and the strain was determined to be Bacillus cereus by morphological, biochemical and molecular biological identification. fermentation culture was carried out in soybean meal, cottonseed meal, soybean meal-cottonseed meal and fish meal fermentation medium respectively, and protease activity and substrate protein digestibility were detected at different times. The results showed that the protease activity and the substrate protein digestibility of strain 2L6 were significantly different in different fermentation medium. Among them, the protease activity in the mixed fermentation medium of soybean meal-cottonseed meal reached a maximum value of 21. 31 bb0. 09U/ m L in a relatively short time, while the digestibility of the substrate protein reached 64. 42% (P0.01) at 36h. Therefore, the mixed protein source of soybean meal-cottonseed meal can greatly improve the protease activity and the ability of protein utilization in the small intestine of piglets. in conclusion, cottonseed meal, corn germ meal and other miscellaneous meal are used as main protein source in daily grain, so that intestinal micro-organism composition of piglets and fattening pigs can be improved, that proportion of pigs can be increased, and the activity of major protease-producing strains in the intestinal tract can be improved, The method can be used as a reasonable alternative to soybean meal in the food.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S828.5
本文编号：2303296
[Abstract]:In order to explore the effect of different protein sources on the intestinal microorganisms in pigs, the effects of the diets on the microbial diversity and the major protease-producing strains of piglets and fattening pigs were studied by using cottonseed meal, fish meal and corn germ meal as the main source of protein. This study consisted of three trials. Effects of different nitrogen sources on small intestinal microorganisms in weanling piglets. Forty-eight weanling piglets (14.62. 1.46 kg) were randomly divided into 4 groups, 3 replicates in each group, 4 piglets were repeated, and the piglets were fed with single cage. The test was treated as control group (SBM group), cottonseed meal (CSM), soybean meal-cottonseed meal (SC) and fish meal (FM) group, among which, the cottonseed meal in the CSM group and SC group was 100% and 50% of the crude protein in the control group respectively. Daily grain is formulated in net energy system to supplement essential amino acids in each group. At the end of the breeding trial, 1 piglets were randomly selected for slaughter and sampling was performed using the Illumina Miseq sequencing technique to determine the diversity of the duodenum, the anterior segment, the middle segment, the posterior segment, and the ileum. The results showed that the diversity of small intestine and the main microbial composition of piglets in different protein source groups were different. Among them, Firmicutes, Proteobacteria, Bacteroidites and Acidobacter were higher in each group, and the abundance of Lactobacillus and Clostridia was higher in the two groups of cottonseed meal, while the abundance of Escherrez/ Shigella in FM group was higher. The results showed that different protein sources had a great influence on the microbial diversity of small intestine of piglets. The effects of different nitrogen sources on the growth and intestinal microorganisms of fattening pigs were studied. Thirty-six (58. 65/ 3. 71 kg) pigs were randomly divided into 4 groups, 3 replicates per process and 3 pigs per replicate. The treatment group was a control group (SBM15) group and three diets (11% CP) diet group, in which 50% of soybean meal (SBM15), cottonseed meal and corn germ meal were replaced by nitrogen instead of soybean meal (SCCM). Cotton seed meal and corn germ meal (100%) were used as the main protein sources in three diets. Daily grain is formulated in net energy system to supplement essential amino acids in each group. After 28 days feeding, ileal contents and feces were taken as intestinal microbial analysis samples, and the microbial diversity of samples was determined by Illumina Miseq sequencing technique. The results showed that the growth performance of SBM15 and CCM was significantly lower than that of SBM15 group (P0.05), but there was no significant difference between SCCM and SBM15 group (P0.05). After the amplification and sequencing of the 16SrRNA V3-V4 region, it was found that the SCCM group day grain significantly increased the microbial diversity of the fattening pigs and increased the abundance of bacteria such as Lactobacillus and Bifidobacterium. In addition, the relative abundance of Rhomoccu, Prevoteellaceae and Bactereoides was also significantly improved in the diet group. Therefore, the soybean meal, the cottonseed meal and the corn germ meal can be used as the main protein source of the daily grain of the fattening pig, the intestinal microorganism can be adjusted, and the normal growth of the fattening pig can be maintained. A protease-producing strain 2L6 was screened from the small intestine of healthy piglets, and the strain was determined to be Bacillus cereus by morphological, biochemical and molecular biological identification. fermentation culture was carried out in soybean meal, cottonseed meal, soybean meal-cottonseed meal and fish meal fermentation medium respectively, and protease activity and substrate protein digestibility were detected at different times. The results showed that the protease activity and the substrate protein digestibility of strain 2L6 were significantly different in different fermentation medium. Among them, the protease activity in the mixed fermentation medium of soybean meal-cottonseed meal reached a maximum value of 21. 31 bb0. 09U/ m L in a relatively short time, while the digestibility of the substrate protein reached 64. 42% (P0.01) at 36h. Therefore, the mixed protein source of soybean meal-cottonseed meal can greatly improve the protease activity and the ability of protein utilization in the small intestine of piglets. in conclusion, cottonseed meal, corn germ meal and other miscellaneous meal are used as main protein source in daily grain, so that intestinal micro-organism composition of piglets and fattening pigs can be improved, that proportion of pigs can be increased, and the activity of major protease-producing strains in the intestinal tract can be improved, The method can be used as a reasonable alternative to soybean meal in the food.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S828.5
【参考文献】
相关期刊论文 前1条
1 房石山;P. Bikker;A. Dirkzwager;J. Fledderus;P. Trevisi;I. le Hueěrou-Luron;J.P.Lallès;A. Awati;;日粮蛋白质和可发酵碳水化合物含量对刚断奶仔猪生长性能和肠道特性的影响[J];国外畜牧学(猪与禽);2007年04期
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