Citric Acid Influences the Dynamics of the Fermentation Quality, Protease Activity and Microbial Community of Mulberry Leaf Silage
Abstract
:1. Introduction
2. Materials and Methods
2.1. Pre-Ensiled Material and Silage Preparation
2.2. Microorganism and Fermentation Parameter Analyses
2.3. Protease Activity Analysis
2.4. Analysis of Microbial Communities
2.5. Statistical Analysis
3. Results and Discussion
3.1. Characteristics of Pre-Ensiled ML
3.2. The Fermentation Parameters, Organic Acids and Microbial Population of ML Silages
3.3. The Protein Fraction of ML Silages
3.4. The Protease Activities of ML Silage
3.5. Bacterial Community of ML Silage
3.5.1. The Dynamic Variation and Alpha Diversity of the Bacterial Community
3.5.2. The Changes in the Dynamics of Relative Abundance among Bacterial Communities
3.5.3. 16S rDNA Gene-Predicted Functional Profiles
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Items | Mulberry Leaves ± SEM |
---|---|
Dry matter (g/kg FM) | 374 ± 12.8 |
Crude protein (g/kg DM) | 139 ± 1.9 |
Neutral detergent fiber (g/kg DM) | 440 ± 35.7 |
Acid detergent fiber (g/kg DM) | 353 ± 24.8 |
Water soluble carbohydrates (g/kg DM) | 114 ± 3.2 |
Lactic acid bacteria (log10 cfu/g FM) | 5.72 ± 0.1 |
Coliform bacteria (log10 cfu/g FM) | 5.21 ± 0.3 |
Yeasts (log10 cfu/g FM) | <2.00 |
Molds (log10 cfu/g FM) | <2.00 |
Item | Treatments | Ensiling Days | SEM | p-Value | |||||
---|---|---|---|---|---|---|---|---|---|
3 | 7 | 14 | 30 | D | T | D × T | |||
Crude protein (g/kg DM) | CK | 134 A,b | 147 A,a | 154 A,a | 153 A,a | 8.57 | 0.000 | 0.000 | 0.011 |
CA1 | 136 A,a | 138 A,B,a | 137 B,a | 146 A,B,a | |||||
CA2 | 133 A,b,c | 130 B,c | 143 B,a | 139 B,a,b | |||||
True protein (g/kg TN) | CK | 878 A,a | 792 A,b | 679 B,c | 620 B,d | 80.1 | 0.000 | 0.679 | 0.000 |
CA1 | 816 B,a | 748 B,b | 752 A,b | 625 A,B,c | |||||
CA2 | 808 B,a | 764 A,B,b | 678 B,c | 690 A,c | |||||
Non-protein nitrogen (g/kg TN) | CK | 122 B,d | 208 B,c | 321 A,b | 380 A,a | 80.1 | 0.000 | 0.679 | 0.000 |
CA1 | 184 A,c | 252 A,b | 248 B,b | 375 A,B,a | |||||
CA2 | 196 A,c | 236 A,b | 322 A,a | 310 B,a | |||||
Ammonia-N (g/kg TN) | CK | 4.40 A | 7.38 A | 14.6 A | 18.9 A | 5.15 | 0.000 | 0.000 | 0.000 |
CA1 | 3.71 B | 4.57 B | 7.58 B | 10.8 B | |||||
CA2 | 1.76 B | 2.45 C | 3.95 C | 6.07 C |
Item | Treatments | Ensiling Days | SEM | p-Value | |||||
---|---|---|---|---|---|---|---|---|---|
3 | 7 | 14 | 30 | D | T | D×T | |||
Carboxypeptidase (units/h/DM) | CK | 70.9 A,b | 66.9 A,b | 80.8 A,a | 83.1 A,a | 7.33 | 0.003 | 0.000 | 0.021 |
CA1 | 69.2 A,a | 74.8 B,a | 76.5 A,a | 80.8 A,a | |||||
CA2 | 66.9 A,a | 66.9 A,a | 66.2 B,a | 65.9 B,a | |||||
Aminopeptidase (units/h/DM) | CK | 86.5 A,a | 83.4 A,a | 23.9 A,b | 17.4 A,b | 28.2 | 0.000 | 0.000 | 0.000 |
CA1 | 59.6 B,a | 65.0 A,a | 18.8 A,B,b | 14.7 A,b | |||||
CA2 | 23.6 C,a | 17.7 B,a,b | 14.75 C,b | 13.5 A,b | |||||
Acid protease (units/h/DM) | CK | 51.3 A,a | 47.1 B,a | 50.5 A,a | 36.6 A,b | 6.76 | 0.000 | 0.018 | 0.360 |
CA1 | 55.2 A,a | 53.9 A,a | 53.5 A,a | 41.3 A,b | |||||
CA2 | 49.7 A,a,b | 55.2 A,a | 54.4 A,a,b | 45.8 A,b |
Item | True Protein | Non-Protein Nitrogen | Ammonia-N | Carboxypeptidase | Aminopeptidase | Acid Protease |
---|---|---|---|---|---|---|
True protein | 1 | |||||
Non-protein nitrogen | −1.00 | 1 | ||||
0.00 | ||||||
Ammonia-N | −0.65 | 0.65 | 1 | |||
0.06 | 0.06 | |||||
Carboxypeptidase | −0.81 | 0.81 | 0.80 | 1 | ||
<0.01 | <0.01 | <0.01 | ||||
Aminopeptidase | −0.51 | 0.51 | 0.56 | 0.62 | 1 | |
0.16 | 0.16 | 0.11 | 0.08 | |||
Acid protease | 0.56 | −0.56 | −0.74 | −0.53 | 0.06 | 1 |
0.11 | 0.11 | 0.02 | 0.14 | 0.87 |
Items | Treatments | Ensiling Days | |||
---|---|---|---|---|---|
3 | 7 | 14 | 30 | ||
Sobs | CK | 1605 | 1339 | 1384 | 1373 |
CA1 | 1477 | 1402 | 1102 | 1305 | |
CA2 | 1376 | 1499 | 1301 | 1337 | |
Ace | CK | 2512 | 2242 | 2263 | 2151 |
CA1 | 2384 | 2348 | 1951 | 2097 | |
CA2 | 2159 | 2417 | 2152 | 2241 | |
Chao1 | CK | 2483 | 2268 | 2204 | 2115 |
CA1 | 2308 | 2294 | 1964 | 2082 | |
CA2 | 2095 | 2453 | 2160 | 2168 | |
Shannon | CK | 3.00 | 3.18 | 5.02 | 5.22 |
CA1 | 2.97 | 4.32 | 4.36 | 4.62 | |
CA2 | 2.62 | 3.59 | 3.68 | 4.13 | |
Simpson | CK | 0.50 | 0.56 | 0.89 | 0.93 |
CA1 | 0.51 | 0.79 | 0.86 | 0.86 | |
CA2 | 0.45 | 0.66 | 0.71 | 0.76 | |
Coverage | CK | 0.99 | 0.99 | 0.99 | 0.99 |
CA1 | 0.99 | 0.99 | 0.99 | 0.99 | |
CA2 | 0.99 | 0.99 | 0.99 | 0.99 |
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Chen, D.; Zhou, W.; Guo, X.; Zheng, M.; Chen, X.; Zhang, Q. Citric Acid Influences the Dynamics of the Fermentation Quality, Protease Activity and Microbial Community of Mulberry Leaf Silage. Fermentation 2021, 7, 185. https://doi.org/10.3390/fermentation7030185
Chen D, Zhou W, Guo X, Zheng M, Chen X, Zhang Q. Citric Acid Influences the Dynamics of the Fermentation Quality, Protease Activity and Microbial Community of Mulberry Leaf Silage. Fermentation. 2021; 7(3):185. https://doi.org/10.3390/fermentation7030185
Chicago/Turabian StyleChen, Dekui, Wei Zhou, Xiang Guo, Mingyang Zheng, Xiaoyang Chen, and Qing Zhang. 2021. "Citric Acid Influences the Dynamics of the Fermentation Quality, Protease Activity and Microbial Community of Mulberry Leaf Silage" Fermentation 7, no. 3: 185. https://doi.org/10.3390/fermentation7030185