不同青贮菌、纤维分解酶及碳源组合对稻秸青贮发酵品质的影响
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摘要
本试验旨在研究不同添加物组合对稻秸青贮发酵品质的影响,进而筛选出最适于稻秸青贮的添加物组合。采用3因素3水平正交设计,将不同种类的青贮菌、纤维分解酶和碳源添加到稻秸中,1组为地衣芽孢杆菌+黑曲霉+葡萄糖,2组为地衣芽孢杆菌+绿色木霉+麸皮,3组为地衣芽孢杆菌+纤维素酶+淀粉,4组为植物乳杆菌+黑曲霉+麸皮,5组为植物乳杆菌+绿色木霉+淀粉,6组为植物乳杆菌+纤维素酶+葡萄糖,7组为粪肠球菌+黑曲霉+淀粉,8组为粪肠球菌+绿色木霉+葡萄糖,9组为粪肠球菌+纤维素酶+麸皮,同时设置空白对照组,共10组,每组3个重复,于青贮15、30和45 d分别采集青贮饲料样品进行青贮发酵品质分析。结果表明:1) 2、4组氨态氮(NH_3-N)/总氮(TN)平均值显著高于空白对照组(P <0.05),9组NH_3-N/TN平均值与空白对照组差异不显著(P>0.05),其他各组均显著低于空白对照组(P<0.05)。2)除1、3组外,其他各组乳酸含量平均值均显著高于空白对照组(P<0.05),除1、3组外,其他各组乙酸含量平均值均显著低于空白对照组(P<0.05),除6、7、8组外,空白对照组丁酸含量平均值显著高于其他各组(P<0.05)。3) 4～9组总评分平均值显著高于空白对照组和其他各组(P<0.05),1、3组总评分平均值与空白对照组没有显著差异(P>0.05)。4)正交分析中,青贮15、30和45 d极差值青贮菌>纤维分解酶,青贮15和45 d极差值纤维分解酶>碳源,且青贮15和30 d都以植物乳杆菌和绿色木霉水平最优,而青贮30和45 d都以麸皮水平最优。综上,对稻秸青贮品质的影响青贮菌>纤维分解酶>碳源,正交筛选最适于稻秸青贮的添加物组合为植物乳杆菌、绿色木霉和麸皮。
The purpose of this experiment was to study the effects of different additives on the fermentation quality of rice straw silage,and then select the most suitable additive combination for rice straw silage. This experiment is an orthogonal test included 3 factors and 3 levels,adding different silage bacteria( Bacillus licheniformis,Lactobacillus plantarum and Enterococcus faecalis),cellulolytic enzymes( Aspergillus niger,Trichoderma viride and cellulase) and carbon source( glucose,bran and starch) to freshly harvested rice straw for silage 45 days,and a blank control group was included. Group 1 was Bacillus licheniformis+Aspergillus niger+glucose,group 2 was Bacillus licheniformis+Trichoderma viride + bran,group 3 was Bacillus licheniformis + cellulase+ starch,group 4 was Lactobacillus plantarum+Aspergillus niger+bran,group 5 was Lactobacillus plantarum+Trichoderma viride+starch,group 6 was Lactobacillus plantarum + cellulase + glucose,group 7 was Enterococcus faecalis + Aspergillus niger +starch,group 8 was Enterococcus faecalis + Trichoderma viride + glucose,and group 9 was Enterococcus faecalis +cellulase+bran. Samples were collected for analyzing the fermentation quality on 15,30 and 45 days of silage.The results showed as follows: 1) the mean of ammoniacal nitrogen( NH_3-N)/total nitrogen( TN) in groups 2 and 4 was significantly higher than that in blank control group( P < 0.05),the mean of NH_3-N/TN in group 9 had no significant difference compared with blank control group( P>0.05),and the mean in the other groups was significantly lower than that in blank control group( P<0.05). 2) Except groups 1 and 3,the mean of lactic acid content in other groups was significantly higher than that in blank control group( P<0.05); except groups 1 and3,the mean of acetic acid content in other groups was significantly lower than that in blank control group( P<0.05); except groups 6,7,8 groups,the mean of butyric acid content in blank control group was significantly higher than that in other groups( P<0.05). 3) The mean of total scores in groups 4 to 9 was significantly higher than that in blank control group and other groups( P<0.05),that in groups 1 and 3 had no significant difference compared with blank control group( P> 0.05). 4) As for orthogonal analysis,the R value was silage bacteria>cellulolytic enzymes at 15,30 and 45 days of silage. At 15 and 45 days of silage,the R value was cellulolytic enzymes>carbon sources,and Lactobacillus plantarum and Trichoderma viride were the excellent level at 15 and 30 days of silage,while bran was the excellent level at 30 and 45 days of silage. In summary,the effect on silage quality is silage bacteria>cellulolytic enzymes>carbon source,and the most suitable additives combination for rice straw silage is Lactobacillus plantarum,Trichoderma viride and bran.
The purpose of this experiment was to study the effects of different additives on the fermentation quality of rice straw silage,and then select the most suitable additive combination for rice straw silage. This experiment is an orthogonal test included 3 factors and 3 levels,adding different silage bacteria( Bacillus licheniformis,Lactobacillus plantarum and Enterococcus faecalis),cellulolytic enzymes( Aspergillus niger,Trichoderma viride and cellulase) and carbon source( glucose,bran and starch) to freshly harvested rice straw for silage 45 days,and a blank control group was included. Group 1 was Bacillus licheniformis+Aspergillus niger+glucose,group 2 was Bacillus licheniformis+Trichoderma viride + bran,group 3 was Bacillus licheniformis + cellulase+ starch,group 4 was Lactobacillus plantarum+Aspergillus niger+bran,group 5 was Lactobacillus plantarum+Trichoderma viride+starch,group 6 was Lactobacillus plantarum + cellulase + glucose,group 7 was Enterococcus faecalis + Aspergillus niger +starch,group 8 was Enterococcus faecalis + Trichoderma viride + glucose,and group 9 was Enterococcus faecalis +cellulase+bran. Samples were collected for analyzing the fermentation quality on 15,30 and 45 days of silage.The results showed as follows: 1) the mean of ammoniacal nitrogen( NH_3-N)/total nitrogen( TN) in groups 2 and 4 was significantly higher than that in blank control group( P < 0.05),the mean of NH_3-N/TN in group 9 had no significant difference compared with blank control group( P>0.05),and the mean in the other groups was significantly lower than that in blank control group( P<0.05). 2) Except groups 1 and 3,the mean of lactic acid content in other groups was significantly higher than that in blank control group( P<0.05); except groups 1 and3,the mean of acetic acid content in other groups was significantly lower than that in blank control group( P<0.05); except groups 6,7,8 groups,the mean of butyric acid content in blank control group was significantly higher than that in other groups( P<0.05). 3) The mean of total scores in groups 4 to 9 was significantly higher than that in blank control group and other groups( P<0.05),that in groups 1 and 3 had no significant difference compared with blank control group( P> 0.05). 4) As for orthogonal analysis,the R value was silage bacteria>cellulolytic enzymes at 15,30 and 45 days of silage. At 15 and 45 days of silage,the R value was cellulolytic enzymes>carbon sources,and Lactobacillus plantarum and Trichoderma viride were the excellent level at 15 and 30 days of silage,while bran was the excellent level at 30 and 45 days of silage. In summary,the effect on silage quality is silage bacteria>cellulolytic enzymes>carbon source,and the most suitable additives combination for rice straw silage is Lactobacillus plantarum,Trichoderma viride and bran.
引文
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[2]GAO L J,YANG H Y,WANG X F,et al.Rice straw fermentation using lactic acid bacteria[J].Bioresource Technology,2008,99(8):2742-2748.
[3]郑会超,杨金勇,应如朗,等.凋萎和添加剂对稻草裹包青贮发酵品质和营养价值的影响[J].动物营养学报,2017,29(4):1312-1318.
[4]QUEIROZ O C M,ARRIOLA K G,DANIEL J L P,et al.Effects of 8 chemical and bacterial additives on the quality of corn silage[J].Journal of Dairy Science,2013,96(9):5836-5843.
[5]BOLSEN K K,ASHBELL G,WEINBERG Z G.Silage fermentation and silage additives-review[J].Asian:Australasian Journal of Animal Sciences,1996,9(5):483-494.
[6]宋鸽,朱小清,张诗,等.绿汁发酵液和纤维素酶对稻草青贮及稻草、甘蔗梢混合青贮品质的影响[J].中国畜牧兽医,2017,44(12):3512-3518.
[7]WANG J,CHEN L,YUAN X,et al.Effects of molasses on the fermentation characteristics of mixed silage prepared with rice straw,local vegetable by-products and alfalfa in Southeast China[J].Journal of Integrative Agriculture,2017,16(3):664-670.
[8]卢强,于浩然,任志花,等.苜蓿青贮添加剂研究进展[J].草原与草业,2018(3):1-4.
[9]郭海明,夏天婵,朱雯,等.青贮添加剂对稻草青贮品质和有氧稳定性的影响[J].草业学报,2017,26(2):190-196.
[10]中国农业部.青贮饲料质量评定标准(试行)[J].饲料工业,1996(21):5-7.
[11]BRODERICK G A,KANG J H.Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media[J].Journal of Dairy Science,1980,63(1):64-75.
[12]AOAC.Official methods of analysis of AOAC international[M].18th ed.Washington,D.C.:AOAC,2005.
[13]KAWAMOTO H,OGAWA M,HAYASAKA K,et al.Effects of formic acid-propionic acid-ammonium complex on fermentation quality and palatability of wrapped round bale silage[J].Bulletin of the National Grassland Research Institute,2000(59):18-23.
[14]KHAN N A,CONE J W,HENDRIKS W H.Stability of fatty acids in grass and maize silages after exposure to air during the feed out period[J].Animal Feed Science and Technology,2009,154(3/4):183-192.
[15]刘瑞江,张业旺,闻崇炜,等.正交试验设计和分析方法研究[J].实验技术与管理,2010,27(9):52-55.
[16]夏友国.产纤维素酶菌株的筛选及不同添加剂在青贮饲料中的应用研究[D].硕士学位论文,福州:福建农林大学,2010.
[17]HOFVENDAHL K,HAHN-HGERDAL B.Factors affecting the fermentative lactic acid production from renewable resources[J].Enzyme and Microbial Technology,2000,26(2/3/4):87-107.
[18]韩吉雨.青贮发酵体系中乳酸菌多样性的研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2009.
[19]KARUNARATNE A,WEZENBERG E,BULLERMANL B.Inhibition of mold growth and aflatoxin production by Lactobacillus spp.[J].Journal of Food Protection,1990,53(3):230-236.
[20]KUNG L,Jr,TAYLOR C C,LYNCH M P,et al.The effect of treating alfalfa with Lactobacillus buchneri40788 on silage fermentation,aerobic stability,and nutritive value for lactating dairy cows[J].Journal of Dairy Science,2003,86(1):336-343.
[21]葛文中,李楠.绿色木霉应用的研究进展[J].黑龙江八一农垦大学学报,2005,17(2):75-80.
[22]VISSERS M M M,DRIEHUIS F,TE GIFFEL M C,et al.Concentrations of butyric acid bacteria spores in silage and relationships with aerobic deterioration[J].Journal of Dairy Science,2007,90(2):928-936.
[23]ZHAO J,DONG Z H,LI J F,et al.Ensiling as pretreatment of rice straw:the effect of hemicellulase and Lactobacillus plantarum on hemicellulose degradation and cellulose conversion[J].Bioresource Technology,2018,266:158-165.
[24]周健民,沈仁芳.土壤学大辞典[M].北京:科学出版社,2013.
[2]GAO L J,YANG H Y,WANG X F,et al.Rice straw fermentation using lactic acid bacteria[J].Bioresource Technology,2008,99(8):2742-2748.
[3]郑会超,杨金勇,应如朗,等.凋萎和添加剂对稻草裹包青贮发酵品质和营养价值的影响[J].动物营养学报,2017,29(4):1312-1318.
[4]QUEIROZ O C M,ARRIOLA K G,DANIEL J L P,et al.Effects of 8 chemical and bacterial additives on the quality of corn silage[J].Journal of Dairy Science,2013,96(9):5836-5843.
[5]BOLSEN K K,ASHBELL G,WEINBERG Z G.Silage fermentation and silage additives-review[J].Asian:Australasian Journal of Animal Sciences,1996,9(5):483-494.
[6]宋鸽,朱小清,张诗,等.绿汁发酵液和纤维素酶对稻草青贮及稻草、甘蔗梢混合青贮品质的影响[J].中国畜牧兽医,2017,44(12):3512-3518.
[7]WANG J,CHEN L,YUAN X,et al.Effects of molasses on the fermentation characteristics of mixed silage prepared with rice straw,local vegetable by-products and alfalfa in Southeast China[J].Journal of Integrative Agriculture,2017,16(3):664-670.
[8]卢强,于浩然,任志花,等.苜蓿青贮添加剂研究进展[J].草原与草业,2018(3):1-4.
[9]郭海明,夏天婵,朱雯,等.青贮添加剂对稻草青贮品质和有氧稳定性的影响[J].草业学报,2017,26(2):190-196.
[10]中国农业部.青贮饲料质量评定标准(试行)[J].饲料工业,1996(21):5-7.
[11]BRODERICK G A,KANG J H.Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media[J].Journal of Dairy Science,1980,63(1):64-75.
[12]AOAC.Official methods of analysis of AOAC international[M].18th ed.Washington,D.C.:AOAC,2005.
[13]KAWAMOTO H,OGAWA M,HAYASAKA K,et al.Effects of formic acid-propionic acid-ammonium complex on fermentation quality and palatability of wrapped round bale silage[J].Bulletin of the National Grassland Research Institute,2000(59):18-23.
[14]KHAN N A,CONE J W,HENDRIKS W H.Stability of fatty acids in grass and maize silages after exposure to air during the feed out period[J].Animal Feed Science and Technology,2009,154(3/4):183-192.
[15]刘瑞江,张业旺,闻崇炜,等.正交试验设计和分析方法研究[J].实验技术与管理,2010,27(9):52-55.
[16]夏友国.产纤维素酶菌株的筛选及不同添加剂在青贮饲料中的应用研究[D].硕士学位论文,福州:福建农林大学,2010.
[17]HOFVENDAHL K,HAHN-HGERDAL B.Factors affecting the fermentative lactic acid production from renewable resources[J].Enzyme and Microbial Technology,2000,26(2/3/4):87-107.
[18]韩吉雨.青贮发酵体系中乳酸菌多样性的研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2009.
[19]KARUNARATNE A,WEZENBERG E,BULLERMANL B.Inhibition of mold growth and aflatoxin production by Lactobacillus spp.[J].Journal of Food Protection,1990,53(3):230-236.
[20]KUNG L,Jr,TAYLOR C C,LYNCH M P,et al.The effect of treating alfalfa with Lactobacillus buchneri40788 on silage fermentation,aerobic stability,and nutritive value for lactating dairy cows[J].Journal of Dairy Science,2003,86(1):336-343.
[21]葛文中,李楠.绿色木霉应用的研究进展[J].黑龙江八一农垦大学学报,2005,17(2):75-80.
[22]VISSERS M M M,DRIEHUIS F,TE GIFFEL M C,et al.Concentrations of butyric acid bacteria spores in silage and relationships with aerobic deterioration[J].Journal of Dairy Science,2007,90(2):928-936.
[23]ZHAO J,DONG Z H,LI J F,et al.Ensiling as pretreatment of rice straw:the effect of hemicellulase and Lactobacillus plantarum on hemicellulose degradation and cellulose conversion[J].Bioresource Technology,2018,266:158-165.
[24]周健民,沈仁芳.土壤学大辞典[M].北京:科学出版社,2013.