You Sihan, Du Shuai, Ge Gentu, Wan Tao, Jia Yushan
Key Laboratory of Forage Cultivation, Processing and High Efficient Utilization, Ministry of Agriculture, Key Laboratory of Grassland Resources, Ministry of Education, College of Grassland, Resources and Environment, Inner Mongolia Agricultural University, Hohhot, China.
National Engineering Laboratory of Biological Feed Safety and Pollution Prevention and Control, Key Laboratory of Molecular Nutrition, Ministry of Education, Key Laboratory of Animal Nutrition and Feed, Ministry of Agriculture and Rural Affairs, Key Laboratory of Animal Nutrition and Feed Science of Zhejiang Province, Institute of Feed Science, Zhejiang University, Hangzhou, China.
Front Microbiol. 2021 Oct 1;12:731770. doi: 10.3389/fmicb.2021.731770. eCollection 2021.
This study aimed to isolate and identify lactic acid bacteria (LAB) from the native grass and naturally fermented silage from the Mongolian Plateau. The effect of selected strains on bacterial community and quality of native grass silage was also studied. Strains XM2, 265, and 842 could grow normally at 15°C-30°C, pH 4.0-8.0, and NaCl 3 and 6.5%; they were identified as subsp. , , and , by sequencing 16S rRNA, respectively. The three strains (XM2, 265, and 842) and one commercial additive (L) were used as inoculants and singularly added to the native grass. Compared to the control, the dry matter content was significantly ( < 0.05) lower in L and XM2 groups. The water-soluble carbohydrate content was significantly ( < 0.05) higher in control than in other groups. Compared with the control, the crude protein and ammonia nitrogen contents were significantly ( < 0.05) higher and lower in the LAB-treated groups, and the acid and detergent fiber contents were significantly ( < 0.05) reduced in the L and XM2 groups than those in other groups. There was a significant ( < 0.05) difference in the pH value, lactic acid content, and lactic acid-to-acetic acid ratio in L and XM2 groups than in other groups. Compared with the control, the number of LAB was significantly ( < 0.05) higher in LAB-treated silages, whereas no significant ( > 0.05) differences were observed in yeast and aerobic bacteria in all groups. Compared to the control, the Shannon index was significantly ( < 0.05) reduced. Simpson and Chao1 were significantly ( < 0.05) increased. Principal coordinate analysis based on the unweighted UniFrac distance showed clear separation of the bacterial community in fresh materials and LAB-treated silages. Besides, compared to the control, the principal coordinate analysis of LAB-treated silages was also separate. After 30 days of fermentation, the relative abundance of increased and was the primary phylum in all silages. Compared with the control, the abundance of and was significantly ( < 0.05) higher and lower in L and XM2 groups. In contrast, no significant differences were observed among control, 265, and 842 groups. At the genus level, the relative abundance of , , , and was increased and dominated the native grass fermentation. Compared with the control, the abundance of was significantly ( < 0.05) higher in L, XM2, and 842 groups, while no significant ( > 0.05) differences were observed between the control and 265 groups. The abundance of was higher than that in other groups. Consequently, the results demonstrated that LAB significantly influenced silage fermentation by reconstructing microbiota, and was the dominant genus in the native grass silages. Furthermore, the results showed that strain XM2 could effectively improve the silage quality, and it is considered a potential starter for the native grass silage.
本研究旨在从蒙古高原的天然牧草及自然发酵青贮饲料中分离和鉴定乳酸菌(LAB)。还研究了所选菌株对天然牧草青贮饲料细菌群落和品质的影响。菌株XM2、265和842在15℃ - 30℃、pH 4.0 - 8.0以及NaCl含量为3%和6.5%的条件下能正常生长;通过对16S rRNA进行测序,它们分别被鉴定为 亚种、 亚种和 亚种。将这三株菌株(XM2、265和842)和一种商业添加剂(L)用作接种剂,单独添加到天然牧草中。与对照组相比,L组和XM2组的干物质含量显著降低(P < 0.05)。对照组的水溶性碳水化合物含量显著高于其他组(P < 0.05)。与对照组相比,LAB处理组的粗蛋白和氨氮含量显著升高和降低(P < 0.05),L组和XM2组的酸性洗涤纤维和中性洗涤纤维含量显著低于其他组(P < 0.05)。L组和XM2组的pH值、乳酸含量以及乳酸与乙酸的比值与其他组相比存在显著差异(P < 0.05)。与对照组相比,LAB处理的青贮饲料中LAB数量显著增加(P < 0.05),而所有组的酵母和好氧细菌数量均未观察到显著差异(P > 0.05)。与对照组相比,香农指数显著降低(P < 0.05)。辛普森指数和Chao1指数显著升高(P < 0.05)。基于非加权UniFrac距离的主坐标分析表明,新鲜物料和LAB处理的青贮饲料中的细菌群落明显分离。此外,与对照组相比,LAB处理的青贮饲料的主坐标分析也相互分离。发酵30天后, 的相对丰度增加,且是所有青贮饲料中的主要门类。与对照组相比,L组和XM2组中 和 的丰度显著升高和降低(P < 0.05)。相比之下,对照组、265组和842组之间未观察到显著差异。在属水平上, 、 、 和 的相对丰度增加,并主导了天然牧草的发酵过程。与对照组相比,L组、XM2组和842组中 的丰度显著升高(P < 0.05),而对照组和265组之间未观察到显著差异(P > 0.05)。 的丰度高于其他组。因此,结果表明LAB通过重建微生物群对青贮饲料发酵有显著影响,且 是天然牧草青贮饲料中的优势属。此外,结果表明菌株XM2能有效提高青贮饲料品质,被认为是天然牧草青贮饲料的潜在发酵剂。