【作者】 曾正清；

【导师】 孙振钧；

【作者基本信息】 中国农业大学 ， 生态学， 2003， 博士

【摘要】 本试验主要研究了日粮中添加乳酸链球菌(Streptococcus lactic)和蚯蚓(Eisenia fetida)粉对猪粪尿中臭气浓度和粘膜免疫机能的影响，以及乳酸菌、蚯蚓粪和牛粪对体外厌氧发酵猪粪尿中臭气浓度的影响。 试验过程：经过对猪粪中分离出的多株乳酸菌菌株进行产酸、耐酸、耐胆盐和耐酶试验，获得了一株产酸能力强，耐酸能力、耐胆盐能力和耐酶能力都较强的一株乳酸链球菌，命名为L4。以L4作为实验用菌株；然后，选用144头杜长大三元杂交仔猪，随机分为四组，以玉米豆粕型日粮为基础日粮，分别添加乳酸菌，蚯蚓粉，乳酸菌+蚯蚓粉，测定其生产性能、粪尿的理化指标、粪尿中正常微生物数量、粪尿中的臭气化合物含量、肠绒毛度、腺窝深度、肠上皮淋巴细胞、杯状细胞以及肥大细胞的数量；同时，直接在猪粪尿中添加乳酸菌、乙酸钠、乳酸菌+乙酸钠、牛粪和蚯蚓粪，32℃下厌氧发酵30天，分别在发酵开始初期、中期和后期采样测定猪粪尿pH、干物质、生物产气量、大肠杆菌、挥发性氨、铵氮、总氮以及粪尿中挥发性脂肪酸、甲酚、吲哚和三甲基吲哚等臭气化合物浓度。 动物试验结果： 对仔猪的影响：1)乳酸菌降低了仔猪粪尿pH、大肠杆菌的数量以及异戊酸、对甲酚含量(P＜0.05)，提高乙酸、丙酸的含量(P＜0.05)；还能增加回肠绒毛长度，小肠各段绒毛长度与腺窝深度的比值，降低回肠的杯状细胞数量和空肠上皮淋巴细胞数量；2)蚯蚓粉能降低仔猪粪尿pH、干物质和粪尿中吲哚的含量(P＜0.05)，能提高十二指肠绒毛长度(P＜0.05)，降低回肠腺窝深度(P＜0.05)，增加血清球蛋白含量的趋势(P＞0.05)，但增加粪尿中乙酸、丙酸和3-甲基吲哚的浓度(P＜0.05)；3)同时添加乳酸菌和蚯蚓粉也明显降低了粪尿中的p-甲酚、吲哚的含量(P＜0.05)，还能够增加十二指肠绒毛长度，绒毛长度与腺窝深度的比例，以及空肠杯状细胞数量(P＜0.05)，降低空肠腺窝深度和肥大细胞的数量(P＜0.05)，而对粪尿pH、挥发性脂肪酸和3-甲基吲哚的含量均无明显影响(P＞0.05)。 对生长猪的影响：1)乳酸菌能降低生长猪粪中大肠杆菌的数量(P＜0.05)，降低对甲酚含量(P＜0.05)；2)蚯蚓粉能降低粪尿干物质和吲哚的含量(P＜0.05)：同时添加乳酸菌和蚯蚓粉能提高了粪尿中挥发性氨的含量(P＜0.05)，但也明显降低了铵氮、大肠杆菌、p-甲酚和吲哚的含量(P＜0.05)，提高粪尿pH、降低了粪尿中乙酸、丙酸、异戊酸、对甲酚和吲哚的含量(P＜0.05)。从以上结果中可看出，乳酸菌提高仔猪的健康水平，很可能是通过减少有害微生物及其他有害因子对肠道的刺激作用，提高动物肠道的健康状况，从而改善了动物肌体健康状态；蚯蚓通过其活性成分增强动物机体的免疫系统，特别是肠道免疫系统的功能，提高动物机体抵抗疾病的能力，从而提高动物的健康状态。 厌氧发酵试验结果： 乳酸菌的作用：乳酸菌能够提高发酵中期猪粪尿中异戊酸的含量(P＜0.05)，降低发酵后期丁酸、3-甲基吲哚的含量(P＜0.05)；同时添加乳酸菌和乙酸钠能明显提高发酵中期丁酸的含量(P＜0.05)；明显降低发酵中期3-甲基吲哚和后期猪粪尿中的丁酸含量(P＜0.05)。 牛粪和蚯蚓粪的作用：牛粪能降低发酵中期猪粪尿中乙酸、丁酸等短链挥发性脂肪酸的浓度 中国农业人学博士学位论义 摘 要 一 （尸＜0刀5），但能增加异成酸、戊酸等链略长挥发性脂肪酸的浓度（尸＜O刀5），随着试验的进行， 异成酸和成酸的浓度才逐渐降低；在所有被测的挥发性脂肪酸中，牛粪只增加了丙酸的浓度，另 外，牛粪还能明显提高猪粪尿的生物产气量（尸＜0．05），降低猪粪尿中3一甲基则保的浓度（尸＜0刀5）； 蚯蚓粪也能降低厌氧发酵早期、中期猪粪尿中3一甲基乳深的浓度（尸＜0刀5），虽然也能降低挥发 性脂肪酸的浓皮，但不如牛粪那么明显，对生物产气量也无明显影响（P＞ 0．05）；牛粪和蚯蚓类 均能提高猪粪尿的PH值（P＜0刀5），同时降低按氮和总氮的含量（P＜0刀5），但对M跺浓度和氨 气的挥发率没有明显影响（N 0乃5）：从以上结果中可看山，牛粪是通过增加猪粪尿中挥发性脂 肪酸转变成生物气体的量，从而减少厌氧发酵的猪粪尿中一些臭气化合物——并发性脂肪酸的含 — — ＆。 从上述试验结果可看出：1）乳酸菌和蚯蚓粉均能提高仔猪的健康水平，乳酸菌和蚯蚓粉同 时添加可降低猪粪尿中臭气化合物的浓度；2）乳酸菌对猪粪尿中的人多数臭气化合物没有明显 的降低作用：牛粪和蚯蚓粪能明显降低厌氧发酵猪粪尿中人多数挥发性脂肪酸的浓度，同时能够 增加生物产气量。更多还原

【Abstract】 In this experiment, effect of lactobacillus (Streptococcus lactic) and earthworm (Eisenia fetidd) supplemented in pig feed on the odorous compounds and pig immunity, and effects of lactobacillus, vermicompost, and cattle feces on the odorous compounds of anaerobically fermented pig slurry were investigated.A strain of lactobacillus was selected and proved to be effective in acid production and have excellent capacity of tolerating acid, bile salt and enzyme as probiotics in the experiment. Four dietary treatments, consisting of the basal diet (CS), basal diet + lactobacillus, basal diet + earthworm and basal diet + lactobacilfus + earthworm, were fed to 144 piglets (Large White * Landrace * Duroc). Growth performance, chemical composition, normal bacteria, odorous compounds in feces, villi height, crypt depth, intraepithelial cell, goblet cell and mast cell were determined in the experiment. Lactobacillus, sodium acetic, lactobacillus + sodium acetic, cattle feces and vermicompost were supplemented in pig slurry fermented anaerobically at 32"C for 30 days, the slurry were sampled for pH, dry mater, biogas production, coliform, ammonia emission, ammonium, total nitrogen, and odorous compounds including volatile fatty acids, p-cresol, indole, and 3-methylindole determination at 10 days, 20 days and 30 days respectively in the experiment.Results from animal trial showed that:Effect on piglets: 1) feces pH, E.coil counts, the concentration of I-valeric acid, p-cresol in the feces, goblet cell in ileum and intraepithelial lymphocytes in jejunum were decreased (P<0.05) by lactobacillus consumption, while the concentration of acetic acid, propionic acid in feces, ileum villi height and the ratio of villi to crypt were increased significantly (P<0.05). 2) Feces DM, pH, the concentration of indole and crypt depth in ileum were decreased significantly (P<0.05), whereas the concentration of acetic, propionic, 3-methylindole in feces, villi height in duodenum and serum globin were increased significantly (P<0.05) by earthworm supplementation. 3) Slurry ammonia emission, serum urea, villi height, ratio of villi to crypt in duodenum and goblet cells in jejunum were increased significantly (P<0.05), slurry p-cresol, indole, jejunum crypt depth and duodenum intraepithelial lymphocytes were decreased significantly (P<0.05), whereas no significant effects on feces pH, volatile fatty acids and 3-methylindole in the feces were observed (P>0.05) when piglets fed with diets containing with both lactobacillus and earthworm.Effect on growers: 1) When the grower diets was supplemented with lactobacillus, p-cresol concentration in the feces were reduced significantly (P<0.05) . 2) Dry material of feces and the indole concentration were decreased significantly (P<0.05 ) , whereas the concentration of 3-methylindole in the feces was increased significantly (P<0.05), when the piglets fed with diets containing earthworm; 3) E.coli counts, the ammonium concentration, acetic, propionic, p-cresol and indole in the feces also were decreased significantly (P<0.05) ; increase of slurry pH, slurry ammonia emission and no significant effect on , other volatile fatty acids and 3-methylindole in the feces (P>0.05) were observed whenpiglets fed with diets containing with both lactobacillus and earthworm. It seems mostly tha lactobacillus enhanced gut health and health status of pig by reducing the impact of harmful bacteria and other harmful factors in feces on pig gastro-intestine, while earthworm bettered the health status of pig by improving the immunity, especially the intestinal mucosal immunity.Results from anaerobic fermentation trial in vitro indicated that:I-valeric concentration in the mid-stages of anaerobic fermentation was significant enhanced by lactobacillus supplementation (P<0.05) butyric concentration during late stage fermentation was lowered significantly by lactobacillus supplementation (P<0.05), Lactobacillus in combination with sodium acetic increased butyric concentration in the mid stage of anaerobic fe更多还原