菜籽饼固态发酵菌株筛选及其发酵产品在猪日粮中的应用

发布时间：2018-04-21 11:34

本文选题：菜籽饼 + 硫代葡萄糖苷　；参考：《安徽农业大学》2015年硕士论文

【摘要】：微生物发酵可提高饲料的利用效率,是目前改善低质蛋白质饲用品质的有效方法之一。目前有关低质蛋白质饲料——菜籽粕的饲用品质改良研究较多,而有关菜籽饼的饲用品质改良研究较少。因此,本研究通过筛选高效硫苷降解菌株,对菜籽饼进行饲用品质的发酵改良,进而通过代谢试验和饲养试验探讨该菌种发酵菜籽饼饲喂猪的营养价值,以及在猪日粮中的最适添加量。1硫苷高效降解菌株的筛选、鉴定和培养条件优化。本研究以菜籽饼厂表层(1-3cm)和深层(3-6cm)土壤为分离源,利用牛肉膏蛋白胨和MRS培养基分离和纯化菌株,分别通过好氧和厌氧筛选得到菌株Y4和R1,再经16S rRNA基因序列分析确定菌株的分类地位。采用单因素和正交试验优化菌株Y4和R1的培养基碳氮源和培养条件。结果显示,Y4和R1分别与蜡样芽孢杆菌(Bacillus cereus)和植物乳杆菌(Lactobacillus plantarum)亲缘关系最近。菌株Y4和R1最优碳源为玉米粉,添加量均为10g/L;最优氮源为发酵芝麻粕,添加量均为15g/L。菌株Y4最优培养条件为初始pH值为7.0,培养温度为37℃,培养时间为24 h,震荡频率为150r/min。菌株R1最优培养条件为初始pH值为6.5,培养温度为35℃,培养时间为32 h。2菜籽饼发酵条件优化。本研究采用单因素和正交试验优化菌株Y4和R1固态好氧发酵条件:接种量、发酵温度、发酵时间、料水比(w/v)和麸皮添加量。结果显示,混菌发酵最优组合为Y4接种量为10%,R1接种量为9%,料水比(w/v)为1:1,麸皮与菜籽饼质量比为1:10(以干物质计),发酵温度为30℃,发酵时间为72 h。本研究混菌发酵效果优于单菌发酵,验证试验测得硫苷、异硫氰酸酯(ITC)和VA唑烷硫酮(OZT)的降解率分别为67.26%、60.38%和53.24%,粗蛋白和小肽含量提高了2.74和2.4个百分点。Y4和R1混合固态发酵菜籽饼能够有效降解硫苷等抗营养因子,可大大改善菜籽饼的饲用品质。3发酵菜籽饼营养价值评定。选择3头健康回肠末端安装“T”型瘘管的杂交阉公猪(杜×长×大,75±1.34 kg),饲喂玉米淀粉—蛋白半纯合日粮,分别以菜籽饼、发酵菜籽饼和酪蛋白作为唯一蛋白源,酪蛋白用于测定内源氮损失,外源指示剂三氧化二铬(Cr2O3)的添加量为0.3%,对菜籽饼发酵前后的营养价值进行评定。结果显示,发酵菜籽饼的消化能(DE)和代谢能(ME)分别为12.11和10.99 MJ/kg,显著高于菜籽饼(P0.05)。发酵菜籽饼的干物质和粗蛋白质的表观消化率较未发酵的菜籽饼分别提高了7.67%和13.66%(P0.05);回肠粗蛋白质表观(真)消化率为79.38%(86.67%),均显著高于菜籽饼(P0.05)。除缬氨酸(Val)、苯丙氨酸(Phe)、精氨酸(Arg)和胱氨酸(Cys)外,发酵菜籽饼中必需氨基酸和非必需氨基酸的回肠表观和真消化率均显著高于菜籽饼(P0.05)。混菌固态发酵有效地改善菜籽饼的饲用品质,显著提高了菜籽饼养分消化利用率。4发酵菜籽饼在生长肥育猪日粮中的应用。选择健康、体重均匀的三元杂交猪(杜×长×大,48±2.32kg)48头,分为3组,每组2圈,每圈8头(3公5母),均采用全自动饲喂系统饲喂,试验日粮采用发酵菜籽饼等能等蛋白替代豆粕。试验猪分为生长阶段(48~65kg)和育肥阶段(66~90kg)。对照组饲喂基础日粮,生长阶段:试验1组发酵菜籽饼替代30%豆粕,试验2组发酵菜籽饼替代60%豆粕。育肥阶段:试验1组发酵菜籽饼替代50%豆粕,试验2组发酵菜籽饼替代100%豆粕。分别检测两阶段猪的生长性能、血清生化指标和肠道大肠杆菌和乳酸菌变化。结果显示,生长阶段发酵菜籽饼替代30%豆粕和育肥阶段发酵菜籽饼替代50%豆粕对猪的生长性能无不良影响,且改善了机体蛋白质和糖代谢及肠道大肠杆菌和乳酸菌组成,促进机体生长发育。发酵菜籽饼能够有效改善菜籽饼的饲用品质,提高了菜籽饼在日粮中的添加比例。
[Abstract]:Microbial fermentation can improve the utilization efficiency of feed and is one of the effective methods to improve the feeding quality of low quality protein. At present, there are many studies on the improvement of the feeding quality of the low quality protein feed - rapeseed meal, and the research on the improvement of the feeding quality of the rapeseed cake is less. The feeding quality of rapeseed cake was improved by fermentation, and then the nutritional value of the rapeseed cake fed by this strain was studied by metabolic test and feeding test, and the optimum addition of.1 glucosinolates in the pig's diet was screened, the identification and culture conditions were optimized. The research was based on the surface (1-3cm) and the deep (3-6cm) soil of the rapeseed cake factory. The soil was separated and purified by beef extract peptone and MRS medium. Strains Y4 and R1 were screened by aerobic and anaerobic screening, and the classification status of the strain was determined by 16S rRNA gene sequence analysis. The medium carbon and nitrogen sources and culture conditions were optimized by single factor and orthogonal test. The results showed that Y4 and R1 scores were found. The closest relationship with Bacillus cereus (Bacillus cereus) and Lactobacillus plantarum (Lactobacillus plantarum). The optimum carbon source of the strain Y4 and R1 is corn flour, the addition amount is 10g/L, the optimal nitrogen source is fermented sesame seed meal. The optimum conditions for the Y4 optimum culture of 15g/L. strain are the initial pH value of 7, the culture temperature of 37, and the culture time of 24. H, the optimal culture condition of 150r/min. strain R1 was the initial pH value of 6.5, the culture temperature of 35 C and the fermentation conditions of 32 H.2 rapeseed cake. The single factor and orthogonal experiment were used to optimize the aerobic fermentation conditions of strain Y4 and R1: inoculation amount, fermentation temperature, fermentation time, feed water ratio (w/v) and bran addition. The results showed that the optimum combination of Y4 inoculation was 10%, R1 inoculation was 9%, the ratio of feed to water (w/v) was 1:1, the mass ratio of bran and rapeseed cake was 1:10 (with dry matter), fermentation temperature was 30, and the fermentation time was 72 h.. The fermentation effect of mixed bacteria was better than single bacteria fermentation, and the test was made of thiocyanate, ITC and OZT (OZT). The degradation rates were 67.26%, 60.38% and 53.24% respectively. The crude protein and small peptide content increased by 2.74 and 2.4 percentage points.Y4 and R1 mixed solid fermented rapeseed cake to effectively degrade the anti nutritional factors such as glucosinolates, which could greatly improve the nutritional value of the rapeseed cake with the feeding quality.3 fermented rapeseed cake. A "T" type fistula was installed at the end of 3 healthy ileum. The hybrid castration pig (DU x long x big, 75 + 1.34 kg) was fed corn starch protein semi homozygous diet with rapeseed cake, fermented rapeseed cake and casein as the only egg white source. Casein was used to determine the loss of endogenous nitrogen. The addition of Cr2O3 was 0.3%. The nutritional value of the rapeseed cake before and after fermentation was carried out. The results showed that the digestion energy (DE) and metabolic energy (ME) of the fermented rapeseed cake were 12.11 and 10.99 MJ/kg, respectively, which were significantly higher than the rapeseed cake (P0.05). The apparent digestibility of the dried rapeseed cake was 7.67% and 13.66% (P0.05) higher than that of the unfermented rapeseed cake, and the apparent digestibility of the ileum crude protein was 79.38% (86.67). Except valine (Val), phenylalanine (Phe), arginine (Arg) and cystine (Cys), the ileum apparent and true digestibility of essential amino acids and non essential amino acids in the fermented rapeseed cake were significantly higher than those of the rapeseed cake (P0.05). The solid fermentation of mixed bacteria effectively improved the feeding quality of the rapeseed cake and significantly improved the vegetable cake (P0.05). The application of seed cake nutrient digestion and utilization rate.4 fermented rapeseed cake in growing fattening pig's diet, the three yuan hybrid pig (DU x long x big, 48 2.32kg) was selected for 48 heads, which were divided into 3 groups, each group was 2 laps, 8 heads per circle (3 male 5 mother), all of them were fed with full automatic feeding system, and the experimental diet used fermented rapeseed cake and other proteins instead of soybean meal. The experimental pigs were divided into the growth stage (48~65kg) and the fattening stage (66~90kg). The control group was fed the basal diet and the growth stage: 1 groups of fermented rapeseed cakes instead of 30% soybean meal, and 2 groups of fermented rapeseed cakes instead of 60% soybean meal. The fattening stage was replaced by 1 groups of rapeseed cakes instead of 50% soybean meal, and 2 groups of fermented rapeseed cakes replaced 100% soybean meal. The two order was detected respectively. The growth performance, serum biochemical indexes and intestinal colibacilli and lactic acid bacteria in the intestine were changed. The results showed that the replacement of 30% soybean meal and 50% soybean meal by the fermentation rapeseed cake in the growing stage had no adverse effect on the growth performance of the pig, and improved the body protein and sugar metabolism, intestinal Escherichia coli and lactic acid bacteria, and promoted the composition of the intestinal bacteria and lactic acid bacteria. The fermented rapeseed cake can effectively improve the feeding quality of rapeseed cake and increase the proportion of rapeseed cake in the diet.

【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828.5

【相似文献】