草业学报 ›› 2022, Vol. 31 ›› Issue (5): 169-177.DOI: 10.11686/cyxb2021091
• 研究论文 • 上一篇
戴东文1,2,3(), 庞凯悦1,2,3(), 王迅1,2,3, 杨英魁1,2,3, 柴沙驼1,2,3(), 王书祥1,2,3()
收稿日期:
2021-03-15
修回日期:
2021-07-12
出版日期:
2022-05-20
发布日期:
2022-03-30
通讯作者:
柴沙驼,王书祥
作者简介:
Corresponding author. E-mail: chaishatuo@163.com, 841271640@qq.com基金资助:
Dong-wen DAI1,2,3(), Kai-yue Pang1,2,3(), xun WANG1,2,3, Ying-kui YANG1,2,3, Sha-tuo CHAI1,2,3(), Shu-xiang WANG1,2,3()
Received:
2021-03-15
Revised:
2021-07-12
Online:
2022-05-20
Published:
2022-03-30
Contact:
Sha-tuo CHAI,Shu-xiang WANG
摘要:
为了研究精料补饲水平对暖季放牧牦牛瘤胃发酵和菌群结构的影响,试验选取体重相近、体况良好的公牦牛48头,随机分为4组,每组12头,以放牧为对照组,3个补饲组分别补饲0.5(Ⅰ)、1.5(Ⅱ)和2.5 kg·d-1(Ⅲ)精料。预试期10 d,正式期60 d。结果表明:1)随着精料补饲水平的提高,瘤胃液pH值,乙酸浓度和乙酸/丙酸值呈线性和二次降低(P<0.05);瘤胃总挥发性脂肪酸、丙酸、丁酸、异丁酸、戊酸和异戊酸浓度呈线性和二次提高(P<0.05);其中补饲Ⅱ和Ⅲ组的微生物蛋白浓度显著高于其他两组(P<0.05)。2)4组共产生3138个OTU,其中共有OTU为1337个,占总OTU数目的42.61%,对照组与补饲Ⅲ组独有OTU分别为183和106个;瘤胃液细菌的物种数、Chao1指数和香农(Shannon)指数随精料补饲的提高呈线性和二次降低(P<0.05)。3)在门水平上,瘤胃拟杆菌门(Bacteroidetes)相对丰度呈线性和二次提高(P<0.05);而厚壁菌门(Firmicutes)相对丰度随精料补饲水平提高呈线性和二次降低(P<0.05);放线菌门(Actinobacteria)相对丰度随精料补饲水平的提高呈线性提高(P<0.05)。在属水平上,随精料补饲水平的提高,瘤胃普雷沃菌属_1(Prevotella_1)、克里斯滕森菌科_R-7(Christensenellaceae_R-7)及普雷沃氏菌科_UCG-001(Prevotellaceae_UCG-001)相对丰度呈线性和二次提高(P<0.05);而解琥珀酸菌属(Succiniclasticum)和瘤胃球菌科_UCG-005(Ruminococcaceae_UCG-005)相对丰度随着精料补饲水平的提高呈线性和二次降低(P<0.05)。综上所述,适宜提高精料补饲水平能够促进暖季放牧牦牛瘤胃发酵,提高瘤胃部分淀粉相关降解菌的丰度。
戴东文, 庞凯悦, 王迅, 杨英魁, 柴沙驼, 王书祥. 精料补饲水平对暖季放牧牦牛瘤胃发酵和菌群结构的影响[J]. 草业学报, 2022, 31(5): 169-177.
Dong-wen DAI, Kai-yue Pang, xun WANG, Ying-kui YANG, Sha-tuo CHAI, Shu-xiang WANG. Effects of different concentrate supplement levels on rumen fermentation and microbial community structure of grazing yaks in the warm season[J]. Acta Prataculturae Sinica, 2022, 31(5): 169-177.
原料 Ingredients | 精料 Concentrate supplement | |
---|---|---|
玉米Zea mays | 44.27 | |
麸皮Wheat bran | 12.06 | |
菜籽粕Rapeseed meal | 12.88 | |
小麦Triticum aestivum | 4.98 | |
豆粕Soybean meal | 12.28 | |
棉籽粕Cottonseed meal | 8.57 | |
碳酸氢钙Ca(HCO3)2 | 1.22 | |
小苏打NaHCO3 | 0.87 | |
氯化钠NaCl | 0.87 | |
预混料Premix1 | 2.00 | |
合计Total | 100.00 | |
营养水平 Nutrient levels2 | 牧草 Pasture | 精料 Concentrate supplement |
粗蛋白质Crude protein (CP) | 11.35 | 18.76 |
粗脂肪Crude fat (EE) | 2.53 | 2.63 |
中性洗涤纤维Neutral detergent fiber (NDF) | 54.16 | 16.03 |
酸性洗涤纤维Acid detergent fiber (ADF) | 26.74 | 7.83 |
钙Calcium (Ca) | 2.15 | 0.60 |
磷Phosphorus (P) | 0.08 | 0.73 |
表1 牧草营养水平、精料组成以及精料营养水平(干物质基础)
Table 1 Nutrient level of the pasture, ingredient and nutrient level of the concentrate supplement (DM basis) (%)
原料 Ingredients | 精料 Concentrate supplement | |
---|---|---|
玉米Zea mays | 44.27 | |
麸皮Wheat bran | 12.06 | |
菜籽粕Rapeseed meal | 12.88 | |
小麦Triticum aestivum | 4.98 | |
豆粕Soybean meal | 12.28 | |
棉籽粕Cottonseed meal | 8.57 | |
碳酸氢钙Ca(HCO3)2 | 1.22 | |
小苏打NaHCO3 | 0.87 | |
氯化钠NaCl | 0.87 | |
预混料Premix1 | 2.00 | |
合计Total | 100.00 | |
营养水平 Nutrient levels2 | 牧草 Pasture | 精料 Concentrate supplement |
粗蛋白质Crude protein (CP) | 11.35 | 18.76 |
粗脂肪Crude fat (EE) | 2.53 | 2.63 |
中性洗涤纤维Neutral detergent fiber (NDF) | 54.16 | 16.03 |
酸性洗涤纤维Acid detergent fiber (ADF) | 26.74 | 7.83 |
钙Calcium (Ca) | 2.15 | 0.60 |
磷Phosphorus (P) | 0.08 | 0.73 |
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析ANOVA | 线性Line | 二次Quadratic | |||
pH | 7.32a | 7.23ab | 6.95b | 6.89b | 0.07 | 0.002 | <0.001 | <0.001 |
微生物蛋白MCP (mg·mL-1) | 2.16c | 2.23c | 2.38b | 2.47a | 0.08 | 0.017 | 0.090 | 0.239 |
氨态氮NH3-N (mg·dL-1) | 11.32a | 10.87ab | 10.65bc | 10.09c | 0.16 | 0.013 | 0.042 | 0.124 |
总挥发性脂肪酸TVFA (mmoL·L-1) | 69.41b | 70.80b | 73.07a | 76.88a | 1.45 | 0.031 | <0.001 | <0.001 |
乙酸Acetic acid (mmoL·L-1) | 51.76a | 49.38a | 44.76b | 42.84c | 1.15 | <0.001 | <0.001 | <0.001 |
丙酸Propionic acid (mmoL·L-1) | 9.73d | 11.89c | 15.46b | 19.91a | 0.51 | <0.001 | <0.001 | <0.001 |
丁酸Butyric acid (mmoL·L-1) | 4.48b | 5.24b | 7.87a | 8.92a | 0.35 | 0.036 | <0.001 | <0.001 |
异丁酸Isobutyric acid (mmoL·L-1) | 0.45b | 0.72b | 0.99a | 1.03a | 0.06 | <0.001 | <0.001 | <0.001 |
戊酸Valeric acid (mmoL·L-1) | 1.35b | 1.71ab | 1.93a | 2.05a | 0.02 | 0.039 | <0.001 | <0.001 |
异戊酸Isovaleric acid (mmoL·L-1) | 1.64c | 1.86b | 2.06a | 2.13a | 0.03 | 0.021 | <0.001 | <0.001 |
乙酸/丙酸Acetic acid/propionic acid | 5.32a | 4.15b | 2.90c | 2.15d | 0.08 | <0.001 | <0.001 | <0.001 |
表2 精料补饲水平对暖季放牧牦牛瘤胃发酵参数的影响
Table 2 Effect of different concentrate supplement levels on rumen fermentation parameters of grazing yaks in the warm season
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析ANOVA | 线性Line | 二次Quadratic | |||
pH | 7.32a | 7.23ab | 6.95b | 6.89b | 0.07 | 0.002 | <0.001 | <0.001 |
微生物蛋白MCP (mg·mL-1) | 2.16c | 2.23c | 2.38b | 2.47a | 0.08 | 0.017 | 0.090 | 0.239 |
氨态氮NH3-N (mg·dL-1) | 11.32a | 10.87ab | 10.65bc | 10.09c | 0.16 | 0.013 | 0.042 | 0.124 |
总挥发性脂肪酸TVFA (mmoL·L-1) | 69.41b | 70.80b | 73.07a | 76.88a | 1.45 | 0.031 | <0.001 | <0.001 |
乙酸Acetic acid (mmoL·L-1) | 51.76a | 49.38a | 44.76b | 42.84c | 1.15 | <0.001 | <0.001 | <0.001 |
丙酸Propionic acid (mmoL·L-1) | 9.73d | 11.89c | 15.46b | 19.91a | 0.51 | <0.001 | <0.001 | <0.001 |
丁酸Butyric acid (mmoL·L-1) | 4.48b | 5.24b | 7.87a | 8.92a | 0.35 | 0.036 | <0.001 | <0.001 |
异丁酸Isobutyric acid (mmoL·L-1) | 0.45b | 0.72b | 0.99a | 1.03a | 0.06 | <0.001 | <0.001 | <0.001 |
戊酸Valeric acid (mmoL·L-1) | 1.35b | 1.71ab | 1.93a | 2.05a | 0.02 | 0.039 | <0.001 | <0.001 |
异戊酸Isovaleric acid (mmoL·L-1) | 1.64c | 1.86b | 2.06a | 2.13a | 0.03 | 0.021 | <0.001 | <0.001 |
乙酸/丙酸Acetic acid/propionic acid | 5.32a | 4.15b | 2.90c | 2.15d | 0.08 | <0.001 | <0.001 | <0.001 |
图1 OTU韦恩图GY、GYSⅠ、GYSⅡ和GYSⅢ分别表示对照组、补饲Ⅰ、Ⅱ和Ⅲ组,下同。GY, GYSⅠ, GYSⅡ and GYSⅢ represent control group, supplementary Ⅰ, Ⅱ and Ⅲ group, the same below.
Fig.1 OTU venn diagram
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析ANOVA | 线性Line | 二次Quadratic | |||
物种数Observed species | 1693.82a | 1499.76ab | 1488.15b | 1294.37c | 26.50 | 0.021 | <0.001 | 0.003 |
Chao1指数Chao1 index | 2091.67a | 1798.12b | 1858.96b | 1650.29b | 55.81 | 0.010 | 0.009 | 0.035 |
Shannon指数Shannon index | 8.61a | 8.53a | 8.37a | 7.80b | 0.10 | <0.001 | <0.001 | <0.001 |
谱系多样性PD whole tree | 137.83a | 128.35a | 125.34a | 113.93a | 2.78 | 0.275 | <0.001 | 0.001 |
覆盖率Coverage (%) | 98.36a | 98.53a | 98.51a | 98.45a | 0.16 | 0.314 | 0.063 | 0.111 |
表3 精料补饲水平对暖季放牧牦牛瘤胃细菌Alpha多样性的影响
Table 3 Effect of different concentrate supplement levels on alpha diversity of rumen bacteria of grazing yaks in the warm season
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析ANOVA | 线性Line | 二次Quadratic | |||
物种数Observed species | 1693.82a | 1499.76ab | 1488.15b | 1294.37c | 26.50 | 0.021 | <0.001 | 0.003 |
Chao1指数Chao1 index | 2091.67a | 1798.12b | 1858.96b | 1650.29b | 55.81 | 0.010 | 0.009 | 0.035 |
Shannon指数Shannon index | 8.61a | 8.53a | 8.37a | 7.80b | 0.10 | <0.001 | <0.001 | <0.001 |
谱系多样性PD whole tree | 137.83a | 128.35a | 125.34a | 113.93a | 2.78 | 0.275 | <0.001 | 0.001 |
覆盖率Coverage (%) | 98.36a | 98.53a | 98.51a | 98.45a | 0.16 | 0.314 | 0.063 | 0.111 |
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析ANOVA | 线性Line | 二次Quadratic | |||
拟杆菌门Bacteroidetes | 53.38c | 59.70b | 65.59a | 68.24a | 1.76 | <0.001 | <0.001 | <0.001 |
厚壁菌门Firmicutes | 39.50a | 33.95b | 25.94c | 25.96c | 1.80 | <0.001 | <0.001 | <0.001 |
放线菌门Actinobacteria | 1.13b | 1.28b | 1.61b | 2.16a | 0.20 | 0.007 | 0.009 | 0.996 |
变形菌门Proteobacteria | 0.55a | 0.86a | 0.61a | 0.57a | 0.07 | 0.431 | 0.455 | 0.479 |
软壁菌门Tenericutes | 1.25a | 1.20a | 0.91a | 0.65a | 0.11 | 0.205 | 0.732 | 0.586 |
疣微菌门Verrucomicrobia | 0.87a | 0.83a | 0.81a | 0.63a | 0.13 | 0.962 | 0.361 | 0.945 |
螺旋体门Spirochaete | 0.53a | 0.45a | 0.19a | 0.21a | 0.06 | 0.091 | 0.231 | 0.068 |
蓝藻菌门Cyanobacteria | 0.51a | 0.43a | 0.39a | 0.36a | 0.07 | 0.079 | 0.133 | 0.317 |
糖细菌门Saccharibacteria | 0.12a | 0.16a | 0.20a | 0.21a | 0.03 | 0.136 | 0.416 | 0.547 |
黏胶球形菌门Lentisphaerae | 0.07a | 0.05a | 0.03a | 0.02a | 0.01 | 0.181 | 0.328 | 0.861 |
表4 精料补饲水平对暖季放牧牦牛瘤胃细菌组成门水平的影响
Table 4 Effect of different concentrate supplement phylum levels on the microbe composition of grazing yaks in the warm season (%)
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析ANOVA | 线性Line | 二次Quadratic | |||
拟杆菌门Bacteroidetes | 53.38c | 59.70b | 65.59a | 68.24a | 1.76 | <0.001 | <0.001 | <0.001 |
厚壁菌门Firmicutes | 39.50a | 33.95b | 25.94c | 25.96c | 1.80 | <0.001 | <0.001 | <0.001 |
放线菌门Actinobacteria | 1.13b | 1.28b | 1.61b | 2.16a | 0.20 | 0.007 | 0.009 | 0.996 |
变形菌门Proteobacteria | 0.55a | 0.86a | 0.61a | 0.57a | 0.07 | 0.431 | 0.455 | 0.479 |
软壁菌门Tenericutes | 1.25a | 1.20a | 0.91a | 0.65a | 0.11 | 0.205 | 0.732 | 0.586 |
疣微菌门Verrucomicrobia | 0.87a | 0.83a | 0.81a | 0.63a | 0.13 | 0.962 | 0.361 | 0.945 |
螺旋体门Spirochaete | 0.53a | 0.45a | 0.19a | 0.21a | 0.06 | 0.091 | 0.231 | 0.068 |
蓝藻菌门Cyanobacteria | 0.51a | 0.43a | 0.39a | 0.36a | 0.07 | 0.079 | 0.133 | 0.317 |
糖细菌门Saccharibacteria | 0.12a | 0.16a | 0.20a | 0.21a | 0.03 | 0.136 | 0.416 | 0.547 |
黏胶球形菌门Lentisphaerae | 0.07a | 0.05a | 0.03a | 0.02a | 0.01 | 0.181 | 0.328 | 0.861 |
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析 ANOVA | 线性 Line | 二次 Quadratic | |||
普雷沃菌属_1 Prevotella_1 | 23.81c | 28.84b | 31.96a | 33.97a | 1.19 | <0.001 | <0.001 | <0.001 |
克里斯滕森菌科_R-7 Christensenellaceae_R-7 | 10.83b | 16.78ab | 16.25ab | 18.67a | 1.15 | 0.032 | 0.001 | 0.002 |
理研菌科_RC9 Gikenellaceae_RC9 | 8.30a | 8.56a | 8.01a | 9.22a | 0.61 | 0.435 | 0.689 | 0.901 |
产粪甾醇真细菌Eubacterium_coprostanoligenes | 3.42a | 3.50a | 3.97a | 4.43a | 0.17 | 0.052 | 0.210 | 0.329 |
解琥珀酸菌属Succiniclasticum | 5.29a | 2.26b | 2.16b | 1.15c | 0.59 | 0.046 | 0.004 | 0.005 |
普雷沃氏菌科_UCG-003 Prevotellaceae_UCG-003 | 1.26a | 1.20a | 1.71a | 1.08a | 0.34 | 0.058 | 0.699 | 0.327 |
瘤胃球菌科_NK4A214 Ruminococcaceae_NK4A214 | 2.95a | 3.08a | 3.39a | 3.24a | 0.32 | 0.087 | 0.134 | 0.295 |
瘤胃球菌科_UCG-005 Ruminococcaceae_UCG-005 | 2.43a | 2.32a | 1.24b | 0.85b | 0.22 | 0.006 | <0.001 | <0.001 |
帕匹杆菌属Papillibacter | 1.07a | 0.74a | 0.80a | 0.91a | 0.11 | 0.381 | 0.401 | 0.769 |
普雷沃氏菌科_UCG-001 Prevotellaceae_UCG-001 | 0.90b | 0.99b | 1.77a | 2.15a | 0.17 | 0.002 | 0.027 | 0.038 |
表5 精料补饲水平对暖季放牧牦牛瘤胃细菌组成属水平的影响
Table 5 Effect of different concentrate supplement genus levels on the microbe composition of grazing yaks in the warm season (%)
项目 Items | 对照组 Control group | 补饲组Supplementary groups | SEM | P值P-value | ||||
---|---|---|---|---|---|---|---|---|
Ⅰ | Ⅱ | Ⅲ | 方差分析 ANOVA | 线性 Line | 二次 Quadratic | |||
普雷沃菌属_1 Prevotella_1 | 23.81c | 28.84b | 31.96a | 33.97a | 1.19 | <0.001 | <0.001 | <0.001 |
克里斯滕森菌科_R-7 Christensenellaceae_R-7 | 10.83b | 16.78ab | 16.25ab | 18.67a | 1.15 | 0.032 | 0.001 | 0.002 |
理研菌科_RC9 Gikenellaceae_RC9 | 8.30a | 8.56a | 8.01a | 9.22a | 0.61 | 0.435 | 0.689 | 0.901 |
产粪甾醇真细菌Eubacterium_coprostanoligenes | 3.42a | 3.50a | 3.97a | 4.43a | 0.17 | 0.052 | 0.210 | 0.329 |
解琥珀酸菌属Succiniclasticum | 5.29a | 2.26b | 2.16b | 1.15c | 0.59 | 0.046 | 0.004 | 0.005 |
普雷沃氏菌科_UCG-003 Prevotellaceae_UCG-003 | 1.26a | 1.20a | 1.71a | 1.08a | 0.34 | 0.058 | 0.699 | 0.327 |
瘤胃球菌科_NK4A214 Ruminococcaceae_NK4A214 | 2.95a | 3.08a | 3.39a | 3.24a | 0.32 | 0.087 | 0.134 | 0.295 |
瘤胃球菌科_UCG-005 Ruminococcaceae_UCG-005 | 2.43a | 2.32a | 1.24b | 0.85b | 0.22 | 0.006 | <0.001 | <0.001 |
帕匹杆菌属Papillibacter | 1.07a | 0.74a | 0.80a | 0.91a | 0.11 | 0.381 | 0.401 | 0.769 |
普雷沃氏菌科_UCG-001 Prevotellaceae_UCG-001 | 0.90b | 0.99b | 1.77a | 2.15a | 0.17 | 0.002 | 0.027 | 0.038 |
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