【作者】 罗燕；

【导师】 闫艳红；

【作者基本信息】 四川农业大学 ， 草学， 2017， 硕士

【摘要】 本试验以多花黑麦草（Lolium multiflorum Lam.）和玉米秸秆为试验材料,探究多花黑麦草与玉米秸秆混贮比例分别为10:0（S0）和7:3（S1）添加植物乳杆菌（Lactobacillus plantment）、布氏乳杆菌（Lactobacillus buchneri）以及纤维素酶对青贮饲料的发酵过程的动态影响。添加剂设6个处理,分别为:对照、B（布氏乳杆菌）、BC（布氏乳杆菌+纤维素酶）、P（植物乳杆菌）、PC（植物乳杆菌+纤维素酶）和BPC（布氏乳杆菌+植物乳杆菌+纤维素酶）,在发酵3d、7d、21d、42d、75d、105d后分析比较不同贮藏时间以及添加剂对青贮料的p H值、发酵品质、V-score评分、营养品质、干物质回收率以及微生物种群的动态影响。结果如下:1.青贮饲料的发酵品质不同添加剂对青贮饲料的发酵品质影响显著（P<0.05）。发酵105d后,S0的p H值、氨态氮/总氮（NH₃-N/TN）比值、挥发性脂肪酸（VFA）显著高于S1,乳酸（LA）含量显著低于S1,S0的V-score评分为12.42分,饲料发酵不良。S0BPC的p H值在发酵7d后降至最低4.2,至发酵105d后p H值升至4.7,LA、AA含量显著高于S0,PA、BA含量、NH₃-N/TN比值显著低于添加植物乳杆菌的处理,但均高于布氏乳杆菌的处理,V-score评分为43.71,饲料发酵不良。S1PC的p H值、NH₃-N/TN比值、VFA含量显著低于其它处理,LA含量显著高于其它处理,其V-score评分为81.83,发酵品质为优;S1BC的p H值在发酵42d后达最低4.7,发酵105d后PA、BA含量、NH₃-N/TN比值显著低于其它处理,AA含量显著高于其它处理,V-score评分78.66。S1BPC的p H值在发酵75d后达最低,发酵105d后PA、BA含量、NH₃-N/TN比值显著低于其它处理,在整个发酵过程中,其LA含量最高,AA含量显著低于添加布氏乳杆菌的处理,V-score评分为83.12,可调制优质的青贮料。2.青贮饲料的化学成分及干物质回收率（DMR）不同添加剂对青贮饲料的化学成分及干物质回收率影响显著（P<0.05）。发酵105d后,S0的可溶性碳水化合物（WSC）、粗蛋白（CP）和干物质（DM）含量显著低于S1,S1的中性洗涤纤维（NDF）和酸性洗涤纤维（ADF）含量显著高于S0。发酵105d后,S0PC较S0显著降低WSC、DM及CP含量,其DMR仅为38.17%,饲料营养成分损失严重。S0BC较S0显著提高WSC、DM含量,显著降低CP、ADF和NDF含量,其DMR仅为52.41%,优于S0PC和S0。S0BPC较S0显著提高WSC、DM含量,降低CP、NDF、ADF含量（差异不显著）,其DMR为49.81%;S1PC较S1显著增加青贮饲料的DM、显著降低WSC、NDF和ADF含量,其DMR为72.07%,饲料保存良好。S1BC较S1显著增加青贮饲料的DM、WSC和CP含量,显著降低NDF、ADF含量,其DMR为77.11%,饲料保存良好。S1BPC较S1显著提高饲料的CP、DM含量,降低WSC、NDF、ADF含量（差异不显著）,其DMR为77.74%,饲料保存良好。3.青贮饲料的微生物的动态变化当多花黑麦草与玉米秸秆的混贮比为10:0时,随贮藏时间的延长,各处理的乳酸菌、酵母菌均呈现先增高后降低的趋势,梭菌数呈递增趋势;当混贮比为7:3时,各处理的乳酸菌、梭菌数变化趋势较弱,酵母菌呈现递减趋势;两个混贮比霉菌数几乎为0,仅在发酵3d后检测到S0霉菌数为10²CFU/g,S1为10⁴CFU/g,S1B为10³CFU/g。饲料在整个贮藏过程中,S0PC的乳酸菌数量在发酵21d后最高,酵母菌数为发酵7d最高,梭菌数最低,随贮藏时间的延长乳酸菌、酵母菌数降低到10⁴ CFU/g、而梭菌的数量增加至10⁶ CFU/g。S1P、S1PC的处理的乳酸菌、酵母菌数量在发酵7d时均达最高,发酵105d后分别逐渐降低到10⁶ CFU/g和10²CFU/g,梭菌的变化趋势较弱。S1B、S1BC的乳酸菌在发酵75d后最高,为10⁶ CFU/g,酵母菌的数量最低,梭菌在整个发酵过程中变化趋势较弱。S1BPC在发酵7～75d后乳酸菌数量基本保持在10⁸ CFU/g,变化趋势较小,梭菌的数量变化较弱,而酵母菌数从发酵3d的10⁶CFU/g逐渐降低到10² CFU/g。综上所述,当多花黑麦草玉米秸秆混贮比为10:0时,各添加剂在发酵前三周表现出良好的效果,但发酵42～105d后梭菌数量增多、青贮料腐败严重,营养成分损失较大。当混贮比为7:3时,各添加剂在整个发酵过程中均起到良好的作用,植物乳杆菌、植物乳杆菌+纤维素酶在发酵前期大量产乳酸使p H值迅速降低,并一直持续到发酵后期,梭菌、酵母菌数量、VFA含量、NH₃-N/TN比值较低,营养成分损失较少。当植物乳杆菌、布氏乳杆菌和纤维素酶混合使用时表现出良好的发酵品质,具有较高的DM、WSC和CP含量,纤维含量较低、干物质回收率高,表现较佳。因此,综合我国南方地区气候条件以及结合实际生产,建议黑麦草玉米秸秆混贮比为7:3时可将3种添加剂混合使用,可调制出优质的多花黑麦草青贮料,若不添加纤维素酶,S1P的发酵品质较佳且保存了更多的化学成分也可进一步推广利用。更多还原

【Abstract】 The Italian ryegrass（Lolium multiflorum Lam.）and Maise straw were the raw material to explore the dynamic effect on fermentation process of two storage proportion of 10:0（S0）and 7:3（S1）and three different addictives which were Lactobacillus plantarum,Lactobacillus buchneri and cellulase.Six treatments were organised in this study which contained control,B（Lactobacillus buchneri）,BC（Lactobacillus buchneri+cellulase）,P（Lactobacillus plantarum）,PC（Lactobacillus plantarum+cellulase）andBPC（Lactobacillus plantarum+Lactobacillus buchneri+cellulase）to analyse the dynamic effects on p H degree,fermentation quality,V-score,nutritional quality,recovery rate of dry matter（DMR）and microbiological composition,which were under the different addictives and ensiling periods of 3d,7d,21d,42d,75d,105d.The results were shown below.1.The fermentation quality of the silageDifferent additives influenced p H degree and fermentation quality of the silage,significantly（P<0.05）.The p H degree,the ratio of ammoniacal nitrogen/total nitrogen（NH₃-N/TN）,volatile fatty acid（VFA）of S0 was higher than S1 and the V-score was 12.42 after ensiled 105 days.The p H degree of S0BPC dropped to the lowest to 4.2after ensiled 7 days and increased to 4.7 after ensiled 105 days,the content of lactic acid（LA）and acetic acid（AA）was higher than S0,the content of PA,BA and NH₃-N/TN were significantly lower than the treatment of Lactobacillus plantarum,but they were higher than the treatment of Lactobacillus buchneri,the V-score was 43.71 and the silage corrupted seriously after ensiled 105 days.The p H degree,NH₃-N/TN and the content of VFA were significantly higher than other treatments,the content of AA was siginificantly higher than other treatments and the V-score was 81.83,the fermentation quality was good.The p H degree of S1BC dropped to the lowest to 4.7 after ensiled 42d and the content of PA,BA and NH₃-N/TN were significantly lower than other treatments,the content of AA was significantly higher than other treatments,the V-score was 78.66.The p H degree of S1BPC dropped to the lowest 4.7 after ensiled 75 days.After ensiled 105 days,the PA,BA and NH₃-N/TN was significantly lower than other treatments.In the whole process of ensiling,the LA content was the highest,the content of AA was lower than the treatment of Lactobacillus buchneri,the V-score was 83.12,witch can be modulated well.2.The chemical components and recovery of dry matter（DMR）of the silageDifferent additives influenced the chemical components and the recovery of dry matter significantly（P<0.05）.After ensiled 105 days,the content of water soluble carbohydrates（WSC）,dry matter（DM）and crude protein（CP）of S0 was siginificantly lower than S1,the content of acid detergent fiber（ADF）and neutral detergent fiber（NDF）of S1 was significantly higher than S0.The DMR was 52.41 witch was better than S0PC and S0.The content of WSC,DM of S0BPC was significantly higher than S0,the DMR was 49.81.The content of DM of S1PC increased significantly and the content of WSC,NDF and ADF reduced significantly,the DMR was 72.07%and the silage was preserved well.The content of DM,WSC and CP of S1BC was increased significantly while the content of ADF,NDF were reduced significantily and the DMR was 77.11%,the silage was preserved well.The content of DM,CP of S1BPC was increased significantly while the content of WSC,NDF and ADF was reduced significantly,the DMR was 77.74%,the silage was preserved well.3.The dynamic change of microorganisms composition of the silageThe number of lactic acid bacteria and yeast of each treatment showed the trend of increasing firstly then decreased and the number of clostridium showed the increasing trend.When the proportion was 7:3,the changing trend of the amount of lactic acid bacteria and clostridium was light while the number of yeast showed a trend of decline.In the whole process of ensiling,the number of lactic acid bacteria of S0PC reached the peak after fermented 21 days,the number of yeast was the highest after fermented 7 days while the number of clostridium was the lowest,the number of yeast,lactic acid bacteria was reduced to 10⁴ CFU/g and the number of clostridium increased to 10⁶CFU/g with the extension of ensiling period.The number of lactic acid bacteria and yeast of S1P and S1PC reached the peak after preserved 7 days,witch were gradually decreased to 10⁶ CFU/g and10²CFU/g,the trend of the number of clostridium changed lightly.The number of lactic acid bacteria of S1BPC maitained the original account of 10⁸ CFU/g between 7 and 75days of fermentation.while the number of clostridium changed lightly from 3 to 75 days of fermentation and the number of yeast reduced from 10⁶ CFU/g to 10² CFU/g.To sum up,when the proportion of Italy ryegrass and Maise straw was 10:0,the additives worked well before three weeks of fermentation.However,it showed bad quality and enormous nutrient loss after ensiled 105 days.When the ratio of Italy ryegrass and Maise straw was 7:3,the additives played a good role in the whole fermentation process.The Lactobacillus plantarum and Lactobacillus plantarum+celluase generated lactic acid quickly in the early stage of the fermentation and reduced the p H degree of the silage and maintained until the later stage of fermentation,the amount of clostridium,yeast and the content of VFA and NH₃-N/TN was lower and the loss of nuritional composition was small.When the three additives were mixed to use,the silage showed the large amount of lactic acid bacteria and small number of harmful bacteria,good fermentation quality and high level of the content of DM,WSC and CP,the less content of fiber and the DMR was higher,witch was better compared with other treatments.Therefore,taking into consideration of regional climate condition comprehensively and combining with the actual practice in south China,we suggested that the ratio of 7:3 witch was combined a mixture of the three additives can produce high-quality silige of Italy ryegrass.S1P can be popularized and utilised futher because of better fermentation quality and less loss of nutrition when the cellulase was removed.更多还原