Effects of two Bacillus species on the micro-silage of cereal straw
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摘要:
利用微生物处理作物秸秆是秸秆饲料化利用的重要途径,对于实现秸秆饲料资源的高效安全利用、促进草食家畜生产具有重要意义。本研究以玉米、水稻和小麦秸秆为研究对象,利用枯草芽孢杆菌和贝莱斯芽孢杆菌及其组合对这三种作物秸秆进行45天发酵试验,以期评价两种芽孢杆菌对不同秸秆纤维降解和微贮效果的影响。结果表明,贝莱斯芽孢杆菌处理对三种秸秆纤维降解有显著影响,与对照组相比,贝莱斯芽孢杆菌使玉米秸秆中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)和酸性洗涤木质素(ADL)分别降低了7.31%、12.0%和21.4%,水稻秸秆分别降低了9.25%、16.0%和43.0%,小麦秸秆ADF下降6.52%;枯草芽孢杆菌处理使玉米秸秆NDF、ADF和ADL分别下降7.19%、13.1%和24.8%;小麦秸秆ADF下降6.96%,但其对水稻秸秆无显著影响,两种芽孢杆菌混合处理没有单菌处理效果显著。两种芽孢杆菌处理秸秆发酵产物中挥发性脂肪酸(VFAs)以乙酸和丁酸为主,未检测到乳酸。贝莱斯芽孢杆菌和枯草芽孢杆菌均能显著降低玉米和小麦秸秆NDF和ADF含量,但由于发酵产物丁酸的存在降低了发酵品质,因此,为了改善微贮质量,未来需探讨两种芽孢杆菌与其它菌种复合效果。
Abstract:Microbial treatment of crop straws is an important approach to feed utilization of straws, which is of great significance for realizing efficient and safe utilization of straws and promoting the production of herbivorous livestock. To evaluate the effects of two kinds of Bacillus on degradation and microsilage of straw fiber, a 45-day fermentation experiment was performed on corn, rice and wheat straws treated with Bacillus subtilis and Bacillus velezensis and their combinations. The results showed that Bacillus velezensis had a significant effect on the degradation of the three fibers, Bacillus velezensis treatment reduced neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL) by 7.31%, 12.0% and 21.4% for corn straw, and 9.25%, 16.0% and 43.0% for rice straw, respectively. ADF of wheat straw by 6.52%. The Bacillus subtilis treatments reduced NDF, ADF and ADL of corn straw by 7.19%, 13.1% and 24.8%, respectively. ADF of wheat straw by 6.96%, but it had no significant effect on rice straw, and the effect of mixed treatment of the two Bacillus species was not as significant as the single Bacillus specie treatment. The fermentation products of straw treated with both Bacillus species were mainly acetic acid and butyric acid, and lactic acid was not detected. Overall, both Bacillus velezensis and Bacillus subtilis could significantly reduce the NDF and ADF contents of corn and wheat straw, but the presence of butyric acid in the fermentation products decreased the fermentation quality. Therefore, in order to enhance the quality of micro-silage of straw, the complex microbial inoculants effect of both Bacillus species with other strains needs to be explored in the future.
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Key words:
- Bacillus velezensis /
- Bacillus subtilis /
- corn straw /
- rice straw /
- wheat straw
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表 1 试验处理
Table 1. Experimental treatment
处理
Treatment组合方式
Composition(A) 对照组
Control group秸秆(50 g)+无菌培养基(18 mL无菌LB培养基∶72 mL无菌水)
Straw (50 g)+ sterile medium (18 mL sterile LB medium∶72 mL sterile water)(B) 贝莱斯芽孢杆菌组
Bacillus velezensis group秸秆(50 g)+贝莱斯芽孢杆菌稀释液(18 mL原液∶72 mL无菌水)
Straw (50 g) + Bacillus velezensis diluent (18 mL stock solution∶72 mL sterile water)(C) 枯草芽孢杆菌组
Bacillus subtilis group秸秆(50 g)+枯草芽孢杆菌稀释液(18 mL原液∶72 mL无菌水)
Straw (50 g) + Bacillus subtilis diluent (18 mL stock solution∶72 mL sterile water)(D) 贝莱斯芽孢杆菌+枯草芽孢杆菌组
Bacillus velezensis and Bacillus subtilis group秸秆(50 g)+混合菌液稀释液(18 mL(1∶1)混合菌液∶72 mL无菌水)
Straw (50 g) + diluent of mixed bacterial solution (18 mL(1∶1) mixed
bacterial solution∶72 mL sterile water)表 2 三种秸秆原料的化学成分
Table 2. Chemical composition of corn, rice and wheat straw
化学成分
Chemical component玉米秸秆
Corn straw水稻秸秆
Rice straw小麦秸秆
Wheat straw干物质 Dry matter/% 93.96 92.87 93.83 总碳 Total carbon/% 42.64 37.45 41.09 总氮 Total nitrogen/% 1.34 2.15 0.77 中性洗涤纤维 Neutral detergent fiber/% 85.13 73.05 88.93 酸性洗涤纤维 Acid detergent fiber/% 53.61 45.32 59.21 酸性洗涤木质素 Acid detergent lignin/% 12.78 6.95 13.54 表 3 缓冲液成分
Table 3. Composition of buffers
g·L−1 缓冲液A
Buffer A含量
Content缓冲液B
Buffer B含量
ContentKH2PO4 10 Na2CO3 15 MgSO4·7H2O 0.5 Na2S·9H2O 1 NaCl 0.5 CaCl2·2H2O 0.1 Urea 0.5 表 4 不同芽孢杆菌条件下三种秸秆的化学组成
Table 4. Chemical composition of three kinds of straw under different Bacillus conditions
玉米秸秆
Corn straw水稻秸秆
Rice straw小麦秸秆
Wheat strawA B C D P A B C D P A B C D P DM/% 33.7±0.16 33.6±0.16 34.3±0.23 34.3±0.34 0.093 32.7±0.09a 31.4±0.42b 31.0±0.12b 32.8±0.08a 0.001 34.0±0.04 34.3±0.01 34.6±0.10 35.4±0.91 0.273 TC/% 43.8±1.04 42.6±0.36 41.9±0.52 43.0±0.07 0.221 40.1±0.15 40.0±0.16 40.0±0.10 39.8±0.05 0.477 42.4±0.15 42.0±0.30 41.8±0.46 42.2±0.12 0.526 TN/% 1.50±0.07 1.52±0.10 1.56±0.12 1.51±0.02 0.962 2.35±0.11a 1.45±0.03b 1.63±0.09b 2.49±0.03a <0.01 0.66±0.05 0.65±0.01 0.62±0.04 0.64±0.02 0.871 NDF/% 86.2±0.68a 79.9±0.61b 80.0±0.61b 84.6±0.81a <0.01 84.3±0.52a 76.5±1.60b 86.8±1.81a 81.6±0.81ab 0.003 76.8±0.59b 79.4±0.55a 80.7±0.42a 80.6±0.41a 0.001 ADF/% 56.5±0.63a 49.7±0.36c 49.1±0.58c 52.8±0.95b <0.01 54.3±0.86a 45.6±0.49c 51.0±0.91ab 50.1±1.08b 0.001 67.5±0.81a 63.1±0.51b 62.8±0.80b 64.7±0.23ab 0.003 ADL/% 14.5±0.86a 11.4±0.04b 10.9±0.61b 13.0±0.53ab 0.01 14.2±1.16a 8.10±0.61b 11.0±0.28ab 11.7±1.09ab 0.007 19.2±0.90 17.6±1.33 19.1±1.15 17.0±1.10 0.470 IVDMD/% 66.1±2.38 62.1±2.08 66.0±0.72 62.9±1.17 0.307 64.1±1.02 65.1±1.41 63.6±0.86 63.8±0.44 0.748 52.0±3.38 50.4±2.17 49.4±0.14 49.7±1.03 0.808 注:A:对照组;B:贝莱斯芽孢杆菌处理组;C:枯草芽孢杆菌处理组;D:两种芽孢杆菌混合处理组;P:判定假设检验结果的参数。DM代表干物质;TC代表总碳;TN代表总氮;NDF代表中性洗涤纤维;ADF代表酸性洗涤纤维;ADL代表酸性洗涤木质素;IVDMD代表体外干物质消化率。不同小写字母代表同种作物秸秆下不同处理间差异达到显著水平(P<0.05)。下同。 Note: A: Control group; B: Bacillus velezensis treatment group; C: Bacillus subtilis treatment group; D: Two kinds of Bacillus mixed treatment group; P: The parameters that determine the results of the hypothesis test. DM indicates dry matter; TC indicates total carbon; TN indicates total nitrogen; NDF indicates neutral detergent fiber;ADF indicates acid detergent fiber;ADL indicates acid detergent lignin; IVDMD indicates In vitro dry matter digestibility. Different small letters indicate significant differences between treatments in the same crop straw at P<0.05. The same is as below. 表 5 不同芽孢杆菌对三种秸秆发酵品质的影响
Table 5. Fermentation quality of three kinds of straw under different Bacillus conditions
玉米秸秆
Corn straw水稻秸秆
Rice straw小麦秸秆
Wheat strawA B C D P A B C D P A B C D P pH 4.61±0.06 4.55±0.05 4.56±0.12 4.44±0.07 0.566 5.07±0.10 4.80±0.06 4.74±0.06 4.89±0.07 0.064 4.77±0.03ab 4.92±0.07a 4.68±0.05b 4.65±0.05b 0.021 铵态氮/总氮
NH3-N/TN /%2.67±0.39 2.47±0.44 2.80±0.37 2.47±0.32 0.177 4.85±0.78a 3.43±0.28b 2.18±0.56c 3.86±0.02b 0.001 4.90±0.85b 6.11±0.21a 3.89±0.46c 3.87±0.30c 0.001 乙酸
Acetic acid/%0.58±0.37 0.43±0.19 0.80±0.66 0.44±0.51 0.741 0.59±0.27 0.86±0.04 0.73±0.28 0.87±0.21 0.413 N.D. 0.51±0.15a 0.19±0.09c 0.40±0.09ab 0.001 丁酸
Butyric acid/%1.05±0.12 1.81±0.29 1.87±1.33 0.91±0.71 0.348 0.43±0.17b 1.12±0.40a 1.18±0.27a 1.18±0.24a 0.031 0.11±0.18b 1.54±0.16a 1.04±0.30a 1.48±0.82a 0.017 注:N.D.代表未检出。 Note: N.D. indicates not detected. -
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