溶菌酶对瘤胃体外发酵、甲烷生成及微生物菌群结构的影响

发布时间：2018-04-28 07:10

本文选题：溶菌酶 + 体外发酵　；参考：《微生物学报》2017年05期

【摘要】：【目的】通过体外静态模拟瘤胃发酵法研究溶菌酶对瘤胃发酵、甲烷生成及微生物菌群结构的影响。【方法】采用单因素多水平试验设计,溶菌酶添加水平分别为0(L-0,对照组)、0.1 mg/100 m L(L-0.1)、1 mg/100 m L(L-1)、10 mg/100 m L(L-10)和100 mg/100 m L(L-100),定时测定产气量和甲烷产量,培养24 h后,发酵液用于发酵参数和微生物菌群数量的q PCR测定,其中L-0、L-1和L-100三个组发酵液同时进行16S r RNA基因Illumina高通量测序。【结果】与对照组相比,低剂量溶菌酶添加(L-0.1组)不影响甲烷产量、氨氮浓度、干物质消失率、有机物消失率和总挥发性脂肪酸等瘤胃发酵参数(P0.05);随着剂量提高,L-1处理组甲烷产量、氨氮浓度显著降低(P0.05),丙酸浓度显著增加(P0.05),并且干物质消失率、有机物消失率和总挥发性脂肪酸不受影响(P0.05);而较高剂量组(L-10和L-100组)虽然甲烷产量显著降低,丙酸浓度显著增加(P0.05),但干物质消失率和有机物消失率也显著降低(P0.05)。q PCR结果显示高剂量组(L-100组)总菌、原虫、甲烷菌数量与对照组相比显著降低(P0.05),而L-0.1、L-1和L-10组总菌、真菌和原虫数量与对照组相比均无显著变化(P0.05)。高通量测序主成分分析(PCA)显示对照组与溶菌酶添加组间瘤胃细菌组成的明显区分,说明添加溶菌酶显著改变了瘤胃细菌菌群结构。溶菌酶通过增加月形单胞菌和琥珀酸弧菌等丙酸生成菌的相对丰度,使更多的氢被用于生成丙酸,导致甲烷产量降低;溶菌酶可抑制普雷沃氏菌和拟杆菌属等蛋白降解菌的生长,进而减少蛋白质过度降解,降低氨氮浓度。【结论】添加适宜浓度(1 mg/100 m L)的溶菌酶可通过调控瘤胃微生态改变瘤胃发酵模式,降低瘤胃甲烷和氨的生成,短期内并不影响饲料消化。
[Abstract]:[objective] to study the effects of lysozyme on rumen fermentation, methane production and microbial flora by static rumen fermentation in vitro. The levels of lysozyme were 0 ~ (-1), 0 ~ (-1) mg/100 路L ~ (-1) and 10 mg/100 ~ (-1) L ~ (-1) and 10 mg/100 / L ~ (-1) L ~ (-1), respectively. The gas production and methane production were measured periodically. After 24 h culture, the fermentation broth was used to determine the fermentation parameters and the number of microflora. The high throughput sequencing of 16s r RNA gene Illumina was carried out in the fermentation broth of L-0 and L-100 groups simultaneously. [results] compared with the control group, low dose lysozyme supplemented with L-0.1 had no effect on methane production, ammonia concentration and dry matter disappearance rate. The rumen fermentation parameters such as organic matter disappearance rate and total volatile fatty acid were P0.05, and with the increase of dosage, the concentration of ammonia nitrogen decreased significantly, the concentration of propionic acid increased P0.05N, and the dry matter disappeared rate. The disappearance rate of organic compounds and total volatile fatty acids were not affected by P0.05N, while the higher dose groups (L-10 and L-100) decreased methane production significantly, The concentration of propionic acid increased significantly, but the disappearance rate of dry matter and organic matter also decreased significantly. The results showed that the total bacteria, protozoa and methane bacteria in high dose group were significantly lower than those in the control group, while the total bacteria in L-0.1 L ~ (-1) and L ~ (10) groups were significantly lower than those in the control group. The number of fungi and protozoa had no significant change compared with the control group (P 0.05). High-throughput sequencing principal component analysis (PCAA) showed that the composition of rumen bacteria was distinguished between the control group and lysozyme added group, which indicated that the addition of lysozyme significantly changed the rumen bacterial flora structure. Lysozyme can increase the relative abundance of propionic acid producing bacteria such as Monomonas monocytogenes and Vibrio succinate so that more hydrogen can be used to produce propionic acid resulting in the reduction of methane production. Lysozyme can inhibit the growth of protein-degrading bacteria, such as Prevos and Bacteroides, and thus reduce the excessive degradation of proteins. [conclusion] the lysozyme supplemented with the appropriate concentration of 1 mg/100 mL can change the rumen fermentation mode and reduce the production of methane and ammonia in the rumen by regulating the rumen microecology, and does not affect feed digestion in the short term.
【作者单位】：江苏省消化道营养与动物健康重点实验室南京农业大学消化道微生物研究室;俄亥俄州立大学动物科学系;
【基金】：国家自然科学基金青年基金(31402101) 江苏省自然科学基金青年基金(BK20140696) 中央高校基本科研业务费专项资金(Y020150023)~~
【分类号】：S816

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