In Vitro Fermentation Characteristics and Methane Mitigation Responded to Flavonoid Extract Levels from Alternanthera sissoo and Dietary Ratios
Abstract
:1. Introduction
2. Material and Methods
2.1. Experiment 1: Preliminary Study
2.1.1. Location and Plantation
2.1.2. Yield Collection and Chemical Analysis
2.1.3. In Vitro Degradability
Animal Ethics
Sample Processing
Ruminal Fluid Donors and Preparation of Inoculum
2.2. Experiment 2: In Vitro Gas Production Technique (Main Experiment)
2.2.1. Flavonoid Extract
2.2.2. Study Design and In Vitro Fermentation
2.2.3. In Vitro Gas Production Sampling and Analysis
3. Statistics
4. Results
4.1. Experiment 1: Preliminary Study
4.1.1. Production of Brazilian Spinach
4.1.2. Chemical Composition and Digestibility of Brazilian Spinach
4.2. Experiment 2: In Vitro Gas Technique (Main Experiment)
4.2.1. Gas Production Kinetics and In Vitro True Dry Matter Digestibility (DMDvt)
4.2.2. Rumen Fermentation, Volatile Fatty Acids (VFAs) and Protozoal Population
5. Discussion
5.1. Brazilian Spinach Yield
5.2. Chemical Composition and Digestibility of Brazilian Spinach
5.3. Gas Production Kinetics and In Vitro Dry Matter Degradability (DMDvt)
5.4. Rumen Fermentation, VFAs and Protozoal Population
6. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Items | Concentrate | Rice Straw |
---|---|---|
Ingredients (g/kg DM) | ||
Cassava chip | 530 | |
Soybean meal | 165 | |
Rice bran | 120 | |
Palm kernel meal | 136 | |
Urea | 10 | |
Premix | 10 | |
Molasses | 14 | |
Sulfur | 5 | |
Salt | 10 | |
Chemical composition | ||
Dry matter (DM), g/kg | 870.7 | 926 |
Crude protein, g/kg DM | 143.8 | 29 |
Neutral detergent fiber, g/kg DM | 216.1 | 716 |
Acid detergent fiber, g/kg DM | 121.4 | 558 |
Ash, g/kg DM | 49.7 | 133 |
Items | Leaf | SEM | p-Value | Leaf + Leaf-Stalk | SEM | p-Value | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
15 | 30 | 45 | 60 | 15 | 30 | 45 | 60 | |||||
Chemical Composition (g/kg) | ||||||||||||
DM | 118.2 b | 126.1 a | 128.2 a | 129.9 a | 1.14 | 0.004 | 148.5 d | 153.2 c | 159.9 b | 168.9 a | 0.14 | 0.001 |
OM | 973.0 b | 973.6 b | 976.7 a | 976.9 a | 0.03 | 0.001 | 958.8 | 958.6 | 957.4 | 955.3 | 0.10 | 0.092 |
CP | 256.4 a | 233.8 b | 214.7 c | 212.0 c | 0.17 | 0.000 | 196.2 a | 170.1 b | 163.0 b | 149.6 c | 0.33 | 0.001 |
NDF | 343.2 b | 303.4 a | 406.1 a | 417.0 a | 0.71 | 0.002 | 477.5 b | 481.9 b | 485.7 b | 497.3 a | 0.35 | 0.019 |
ADF | 173.8 c | 192.7 b | 200.6 b | 210.9 a | 0.33 | 0.002 | 207.1 | 209.9 | 212.1 | 212.9 | 0.68 | 0.202 |
TF (g/kg) | 82.16 a | 72.21 b | 50.73 c | 49.57 c | 0.48 | 0.000 | 78.14 a | 71.34 b | 50.04 c | 45.52 d | 1.06 | 0.000 |
Items (g/kg) | Leaf | SEM | p-Value | Leaf and Leaf + Leaf-Stalk | SEM | p-Value | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
15 | 30 | 45 | 60 | 15 | 30 | 45 | 60 | |||||
12 h | ||||||||||||
DM | 559.4 | 556.0 | 551.4 | 542.1 | 0.70 | 0.220 | 468.5 a | 406.4 b | 366.0 c | 346.6 d | 0.37 | 0.000 |
OM | 871.2 a | 863.9 b | 857.0 c | 839.2 a | 0.76 | 0.000 | 862.2 a | 844.1 b | 841.9 b | 839.9 b | 0.15 | 0.002 |
NDF | 398.1 a | 389.3 a | 364.9 b | 358.7 b | 0.43 | 0.002 | 341.6 a | 312.6 b | 262.6 c | 234.0 d | 0.07 | 0.000 |
ADF | 229.0 a | 218.1 b | 207.6 c | 186.0 d | 0.32 | 0.001 | 177.1 a | 134.7 b | 130.1 c | 129.2 c | 0.04 | 0.000 |
24 h | ||||||||||||
DM | 629.4 a | 615.8 b | 611.0 c | 602.8 d | 0.08 | 0.000 | 606.0 a | 600.0 b | 574.6 c | 542.7 d | 0.07 | 0.000 |
OM | 934.6 a | 927.6 b | 912.7 c | 900.6 d | 0.07 | 0.000 | 906.0 a | 899.4 b | 881.7 c | 879.2 d | 0.07 | 0.000 |
NDF | 458.8 a | 457.2 a | 449.3 b | 438.9 c | 0.07 | 0.000 | 423.1 a | 396.9 b | 385.5 bc | 374.7 c | 0.71 | 0.009 |
ADF | 259.3 a | 257.4 a | 246.6 b | 238.8 c | 0.07 | 0.000 | 247.0 a | 224.8 b | 212.2 c | 207.0 d | 0.07 | 0.000 |
Treatments | R:C | FE (mg) | a (mL) | B (mL) | c (mL) | a + b (mL) | Gas Production at 96 h (mL/0.5 g Substrate) | DMDvt (%) | |
---|---|---|---|---|---|---|---|---|---|
12 h | 24 h | ||||||||
T1 | 50:50 | 0 | −6.17 | 109.17 | 0.039 | 103.03 | 93.1 | 51.72 | 63.65 |
T2 | 10 | −3.1 | 127.8 | 0.02 | 124.7 | 99.5 | 51.58 | 65.2 | |
T3 | 20 | −1.6 | 116.9 | 0.027 | 115.43 | 102.7 | 51.13 | 67.9 | |
T4 | 30 | −2.3 | 110.13 | 0.024 | 107.93 | 88.9 | 46.93 | 68.52 | |
T5 | 40 | −1.33 | 107.43 | 0.078 | 106.03 | 101.7 | 57.23 | 68.9 | |
T6 | 40:60 | 0 | −1.3 | 117.53 | 0.019 | 116.4 | 105.4 | 60 | 62.98 |
T7 | 10 | −4.23 | 121.07 | 0.018 | 116.83 | 94.4 | 62.33 | 72.25 | |
T8 | 20 | −5.53 | 111.93 | 0.032 | 108.4 | 100.9 | 61.22 | 70.48 | |
T9 | 30 | −4.2 | 130.6 | 0.019 | 126.4 | 101.5 | 65.05 | 72.02 | |
T10 | 40 | −2 | 107.7 | 0.018 | 105.7 | 82.4 | 64.77 | 71.6 | |
T11 | 30:70 | 0 | −1.25 | 126 | 0.038 | 124.75 | 113.1 | 68.8 | 78.3 |
T12 | 10 | −2.07 | 119.53 | 0.022 | 117.7 | 98.7 | 67.28 | 79.37 | |
T13 | 20 | −3.4 | 112.75 | 0.021 | 119.3 | 95.6 | 70.2 | 86.17 | |
T14 | 30 | −3.57 | 100.5 | 0.03 | 97 | 88.7 | 74.57 | 84.88 | |
T15 | 40 | −3.1 | 113.73 | 0.028 | 110.63 | 97.3 | 70.6 | 74.8 | |
SEM | 3.098 | 17.87 | 0.01 | 15.9 | 14.18 | 12.46 | 18 | ||
R:C | 50:50 | −1.29 | 114.31 | 0.029 | 111.41 | 97.2 | 51.72 b | 66.84 b | |
40:60 | −2.88 | 112.17 | 0.027 | 114.55 | 94.51 | 62.68 ab | 69.86 b | ||
30:70 | −2.89 | 111.71 | 0.028 | 110.41 | 98.71 | 68.83 a | 80.73 a | ||
FE (mg) | 0 | −1 | 113.12 | 0.034 a | 112.13 | 103.87 | 60.18 | 68.31 | |
10 | −3.04 | 122.80 | 0.020 b | 119.73 | 97.54 | 60.40 | 72.28 | ||
20 | −2.43 | 113.64 | 0.028 ab | 111.2 | 99.73 | 60.85 | 74.84 | ||
30 | −3.36 | 113.78 | 0.024 ab | 110.43 | 93.07 | 59.75 | 75.14 | ||
40 | −1.93 | 100.29 | 0.032 a | 107.13 | 89.81 | 64.20 | 71.80 | ||
Orthogonal polynomial | Lin | 0.510 | 0.075 | 0.985 | 0.259 | 0.621 | 0.651 | 0.678 | |
Quad | 0.172 | 0.107 | 0.030 | 0.485 | 0.620 | 0.722 | 0.548 | ||
Cub | 0.925 | 0.784 | 0.359 | 0.424 | 0.264 | 0.746 | 0.924 | ||
R:C × FE | 0.061 | 0.234 | 0.832 | 0.533 | 0.153 | 0.999 | 0.999 |
Treatments | R:C | FE (mg) | pH | NH3-N (mg/dL) | Protozoa (×105 Cells/mL) |
---|---|---|---|---|---|
T1 | 50:50 | 0 | 6.85 | 10.82 | 3.50 |
T2 | 10 | 6.87 | 17.03 | 3.25 | |
T3 | 20 | 6.91 | 10.63 | 3.00 | |
T4 | 30 | 6.94 | 17.13 | 1.25 | |
T5 | 40 | 6.91 | 17.92 | 1.50 | |
T6 | 40:60 | 0 | 6.61 | 20.02 | 3.50 |
T7 | 10 | 6.66 | 18.43 | 2.75 | |
T8 | 20 | 6.60 | 20.63 | 3.25 | |
T9 | 30 | 6.64 | 16.52 | 2.75 | |
T10 | 40 | 6.65 | 21.42 | 2.00 | |
T11 | 30:70 | 0 | 6.49 | 22.92 | 5.50 |
T12 | 10 | 6.40 | 22.22 | 4.52 | |
T13 | 20 | 6.50 | 24.32 | 3.50 | |
T14 | 30 | 6.41 | 22.22 | 4.00 | |
T15 | 40 | 6.60 | 21.82 | 1.87 | |
SEM | 0.013 | 0.255 | 0.052 | ||
R:C | 50:50 | 6.90 a | 14.71 b | 2.50 b | |
40:60 | 6.63 b | 19.41 b | 2.85 b | ||
30:70 | 6.48 b | 22.70 a | 3.88 a | ||
FE (mg) | 0 | 6.65 | 17.92 | 4.17 a | |
10 | 6.65 | 19.23 | 3.51 ab | ||
20 | 6.67 | 18.53 | 3.25 ab | ||
30 | 6.66 | 18.62 | 2.67 bc | ||
40 | 6.72 | 20.39 | 1.79 b | ||
Orthogonal polynomial | Lin | 0.556 | 0.394 | 0.000 | |
Quad | 0.773 | 0.773 | 0.534 | ||
Cub | 0.895 | 0.467 | 0.502 | ||
R:C × FE | 0.995 | 0.437 | 0.299 |
Treatments | R:C | FE (mg) | C2:C3 Ratio | Molar Proportions of VFA (mmol/L) | Total VFA (mmol/L) | CH4 (mmol/L) | ||
---|---|---|---|---|---|---|---|---|
C2 | C3 | C4 | ||||||
T1 | 50:50 | 0 | 4.14 a | 74.75 | 18.06 | 10.43 | 94.85 | 24.96 |
T2 | 10 | 3.92 a | 72.44 | 18.46 | 10.62 | 94.53 | 21.99 | |
T3 | 20 | 3.08 bc | 72.59 | 24.24 | 10.71 | 102.12 | 20.93 | |
T4 | 30 | 3.17 bcd | 72.65 | 24.48 | 12.12 | 107.31 | 20.30 | |
T5 | 40 | 3.86 a | 72.28 | 20.16 | 10.56 | 99.09 | 19.59 | |
T6 | 40:60 | 0 | 3.21 b | 69.97 | 21.79 | 12.51 | 104.28 | 19.42 |
T7 | 10 | 2.87 bcde | 68.36 | 23.86 | 11.41 | 103.62 | 16.30 | |
T8 | 20 | 3.25 bc | 68.18 | 21.57 | 11.01 | 102.76 | 11.55 | |
T9 | 30 | 2.84 bcde | 66.24 | 24.52 | 10.3 | 104.55 | 10.74 | |
T10 | 40 | 2.83 bcde | 63.48 | 25.62 | 12.02 | 110.13 | 10.04 | |
T11 | 30:70 | 0 | 2.8 bcde | 66.35 | 23.73 | 11.52 | 109.99 | 16.29 |
T12 | 10 | 2.65 de | 65.45 | 24.73 | 11.71 | 108.88 | 13.25 | |
T13 | 20 | 2.43 e | 65.34 | 27.66 | 11.84 | 114.08 | 12.55 | |
T14 | 30 | 2.66 cde | 65.22 | 26.58 | 12.59 | 116.81 | 11.91 | |
T15 | 40 | 2.49 e | 64.87 | 27.5 | 12.61 | 117.89 | 10.82 | |
SEM | 0.25 | 0.100 | 1.93 | 1.076 | 4.25 | 0.156 | ||
R:C | 50:50 | 3.64 a | 72.94 a | 21.22 c | 10.89 b | 99.61 c | 21.55 a | |
40:60 | 3.02 b | 67.25 b | 23.47 b | 11.45 ab | 104.87 b | 13.61 b | ||
30:70 | 2.61 c | 65.45 c | 26.24 a | 12.06 a | 113.53 a | 12.97 b | ||
FE (mg) | 0 | 3.41 a | 70.36 ab | 20.59 b | 11.49 | 103.04 b | 20.22 a | |
10 | 3.15 ab | 68.75 ab | 22.87 ab | 11.25 | 101.44 b | 17.18 b | ||
20 | 2.94 b | 68.70 ab | 24.49 a | 11.19 | 106.38 ab | 15.01 bc | ||
30 | 2.89 b | 68.04 b | 25.19 a | 11.67 | 109.56 a | 14.32 bc | ||
40 | 3.06 b | 66.88 b | 24.43 a | 11.74 | 109.04 a | 13.48 c | ||
Orthogonal polynomial | Lin | 0.010 | 0.001 | 0.002 | 0.523 | 0.003 | 0.000 | |
Quad | 0.019 | 0.904 | 0.120 | 0.493 | 0.932 | 0.121 | ||
Cub | 0.595 | 0.308 | 0.265 | 0.670 | 0.091 | 0.743 | ||
R:C × FE | 0.042 | 0.356 | 0.122 | 0.398 | 0.483 | 0.785 |
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Sommai, S.; Cherdthong, A.; Suntara, C.; So, S.; Wanapat, M.; Polyorach, S. In Vitro Fermentation Characteristics and Methane Mitigation Responded to Flavonoid Extract Levels from Alternanthera sissoo and Dietary Ratios. Fermentation 2021, 7, 109. https://doi.org/10.3390/fermentation7030109
Sommai S, Cherdthong A, Suntara C, So S, Wanapat M, Polyorach S. In Vitro Fermentation Characteristics and Methane Mitigation Responded to Flavonoid Extract Levels from Alternanthera sissoo and Dietary Ratios. Fermentation. 2021; 7(3):109. https://doi.org/10.3390/fermentation7030109
Chicago/Turabian StyleSommai, Sukruthai, Anusorn Cherdthong, Chanon Suntara, Sarong So, Metha Wanapat, and Sineenart Polyorach. 2021. "In Vitro Fermentation Characteristics and Methane Mitigation Responded to Flavonoid Extract Levels from Alternanthera sissoo and Dietary Ratios" Fermentation 7, no. 3: 109. https://doi.org/10.3390/fermentation7030109