Effects of the Appropriate Addition of Antioxidants from Pinus densiflora and Mentha canadensis Extracts on Methane Emission and Rumen Fermentation
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. In Vitro Batch Fermentation
2.3. Total Phenolic and Flavonoid Content of Pinus densiflora and Mentha canadensis Extracts
2.4. Total DPPH, ABTS and HO, NO Radical Assay
2.5. Analysis of In Vitro End-Products
2.6. Relative Quantification of Specific Ruminal Microbes
2.7. Statistical Analysis
3. Results
3.1. Total Phenol and Flavonoid Contents, DPPH and ABTS Assays, OH and NO Scavenging Activities
3.2. Effect of Terpene-Based Plant Extracts on Rumen Fermentation and Dry Matter Degradability
3.3. Effect of Terpene-Based Plant Extracts on Gas Profiles
3.4. Effect of Terpene-Based Plant Extracts on VFA Profiles
3.5. Relative Quantification of Specific Ruminal Microbes
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Content | Terpene-Based Plant Extract | |
---|---|---|
Pinus densiflora | Mentha canadensis | |
Total polyphenol content (µg GAE/g) | 925 ± 113.24 | 469 ± 14.27 |
Total flavonoid content (µg QE/g) | 680.90 ± 67.49 | 715.77 ± 61.01 |
Total antioxidant capacity | ||
DPPH | ||
200 (µg/mL) | 49.38 ± 2.54 | 48.25 ± 1.28 |
100 (µg/mL) | 43.27 ± 2.66 | 47.56 ± 1.63 |
50 (µg/mL) | 21.90 ± 1.09 | 37.95 ± 4.63 |
10 (µg/mL) | 16.63 ± 3.39 | 17.38 ± 1.53 |
ABTS | ||
200 (µg/mL) | 78.85 ± 0.98 | 91.36 ± 0.29 |
100 (µg/mL) | 67.34 ± 0.85 | 90.51 ± 0.68 |
50 (µg/mL) | 52.20 ± 3.41 | 82.68 ± 1.20 |
10 (µg/mL) | 23.11 ± 1.16 | 79.50 ± 1.97 |
HO | ||
200 (µg/mL) | 2336.36 ± 100.39 | 299.97 ± 43.00 |
100 (µg/mL) | 2251.17 ± 137.16 | 179.81 ± 30.84 |
50 (µg/mL) | 1707.47 ± 115.93 | 102.46 ± 7.81 |
10 (µg/mL) | 790.53 ± 50.97 | 37.11 ± 6.80 |
NO | ||
200 (µg/mL) | 59.31 ± 1.36 | 54.93 ± 2.92 |
100 (µg/mL) | 32.42 ± 2.01 | 32.50 ± 2.52 |
50 (µg/mL) | 20.59 ± 3.14 | 20.35 ± 3.71 |
10 (µg/mL) | 7.94 ± 2.59 | 12.04 ± 3.74 |
Incubation Time (h) | Treatment | Extract Concentration (1) (mg/L) | SEM (2) | p Value (3) | |||||
---|---|---|---|---|---|---|---|---|---|
0 | 30 | 50 | 70 | T | L | Q | |||
pH | |||||||||
12 | Herb (4) | 6.58 a | 6.60 a | 6.48 b | 6.62 a | 0.01 | 0.0003 | 0.7580 | 0.0099 |
Pine (5) | 6.79 a | 6.80 a | 6.69 b | 6.82 a | 0.02 | 0.0055 | 0.7644 | 0.0338 | |
24 | Herb | 6.24 | 6.53 | 6.49 | 6.51 | 0.11 | 0.2922 | 0.1268 | 0.3111 |
Pine | 6.70 a | 6.73 a | 6.56 b | 6.71 a | 0.03 | 0.0136 | 0.2929 | 0.2027 | |
Microbial growth rate, OD at 550 nm | |||||||||
12 | Herb | 0.42 a | 0.30 b | 0.41 a | 0.39 ab | 0.03 | 0.0792 | 0.7914 | 0.0901 |
Pine | 0.34 | 0.34 | 0.40 | 0.35 | 0.06 | 0.8686 | 0.7922 | 0.7290 | |
24 | Herb | 0.35 ab | 0.37 ab | 0.37 a | 0.31 b | 0.02 | 0.1179 | 0.2543 | 0.0433 |
Pine | 0.35 | 0.29 | 0.38 | 0.35 | 0.03 | 0.1862 | 0.5431 | 0.3383 | |
Glucose (mg∙mL−1) | |||||||||
12 | Herb | 0.07 a | 0.06 bc | 0.06 c | 0.07 ab | 0.00 | 0.0244 | 0.1329 | 0.0086 |
Pine | 0.06 | 0.08 | 0.06 | 0.05 | 0.01 | 0.2376 | 0.1892 | 0.1667 | |
24 | Herb | 0.05 b | 0.07 a | 0.05 b | 0.06 ab | 0.01 | 0.0242 | 0.2306 | 0.0446 |
Pine | 0.06 | 0.06 | 0.05 | 0.05 | 0.01 | 0.3502 | 0.1135 | 0.4862 | |
Protein (mg∙mL−1) | |||||||||
12 | Herb | 0.20 a | 0.17 ab | 0.18 ab | 0.16 b | 0.01 | 0.0962 | 0.0273 | 0.8446 |
Pine | 0.20 | 0.20 | 0.23 | 0.21 | 0.01 | 0.1537 | 0.2807 | 0.4698 | |
24 | Herb | 0.19 b | 0.19 b | 0.20 ab | 0.21 a | 0.01 | 0.0473 | 0.0105 | 0.2879 |
Pine | 0.22 | 0.22 | 0.23 | 0.21 | 0.01 | 0.3240 | 0.7081 | 0.1946 | |
Ammonia nitrogen (mg∙mL−1) | |||||||||
12 | Herb | 3.18 a | 3.93 a | 1.18 b | 2.93 a | 0.44 | 0.0123 | 0.1272 | 0.6570 |
Pine | 3.98 | 7.78 | 4.16 | 6.33 | 1.15 | 0.1324 | 0.4231 | 0.3784 | |
24 | Herb | 2.60 ab | 2.27 b | 4.31 a | 2.89 ab | 0.52 | 0.0983 | 0.2542 | 0.5618 |
Pine | 4.80 b | 4.78 b | 5.51 ab | 6.53 a | 0.46 | 0.0854 | 0.0260 | 0.1910 |
Incubation Time (h) | Treatment | Extract Concentration (1) (mg/L) | SEM (2) | p Value (3) | |||||
---|---|---|---|---|---|---|---|---|---|
0 | 30 | 50 | 70 | T | L | Q | |||
DM degradability, % | |||||||||
3 | Herb (4) | 19.33 | 18.67 | 17.89 | 19.56 | 0.76 | 0.4468 | 0.8850 | 0.1900 |
Pine (5) | 22.11 | 21.22 | 23.00 | 21.33 | 1.29 | 0.7519 | 0.9133 | 0.8601 | |
6 | Herb | 20.11 | 19.78 | 19.00 | 19.67 | 1.14 | 0.9159 | 0.6732 | 0.7375 |
Pine | 23.00 | 22.00 | 23.11 | 22.22 | 0.82 | 0.7196 | 0.7093 | 0.8722 | |
9 | Herb | 21.44 | 21.22 | 21.22 | 20.00 | 1.01 | 0.7416 | 0.3842 | 0.5983 |
Pine | 23.00 | 23.89 | 23.44 | 23.11 | 1.14 | 0.9444 | 0.9673 | 0.5882 | |
12 | Herb | 27.11 | 26.33 | 27.11 | 27.33 | 0.38 | 0.332 | 0.5261 | 0.1647 |
Pine | 29.89 | 29.78 | 29.11 | 31.00 | 0.85 | 0.505 | 0.5529 | 0.2887 | |
24 | Herb | 30.78 | 33.78 | 34.67 | 29.44 | 2.00 | 0.2816 | 0.8716 | 0.0784 |
Pine | 32.44 | 33.22 | 31.44 | 33.33 | 1.25 | 0.6992 | 0.8741 | 0.7543 | |
48 | Herb | 41.22 | 38.33 | 39.78 | 33.78 | 1.77 | 0.0776 | 0.0314 | 0.3655 |
Pine | 36.89 | 37.67 | 39.11 | 36.56 | 0.97 | 0.3167 | 0.8298 | 0.1564 | |
72 | Herb | 49.37 | 52.46 | 50.79 | 50.96 | 1.43 | 0.5362 | 0.5485 | 0.3099 |
Pine | 48.67 | 48.56 | 55.22 | 51.89 | 1.63 | 0.0594 | 0.0568 | 0.6463 |
Incubation Time (h) | Treatment | Extract Concentration (1) (mg/L) | SEM (2) | p Value (3) | |||||
---|---|---|---|---|---|---|---|---|---|
0 | 30 | 50 | 70 | T | L | Q | |||
Total gas, mL/g DM | |||||||||
12 | Herb (4) | 190.27 b | 189.64 b | 204.58 a | 187.32 b | 2.01 | 0.0011 | 0.4233 | 0.0094 |
Pine (5) | 167.94 ab | 163.24 bc | 175.01 a | 159.49 c | 2.37 | 0.0088 | 0.2539 | 0.0950 | |
24 | Herb | 212.13 | 207.75 | 214.03 | 205.00 | 4.75 | 0.5535 | 0.4962 | 0.6923 |
Pine | 176.97 b | 177.92 b | 187.32 a | 176.97 b | 2.79 | 0.0811 | 0.4221 | 0.1367 | |
Methane, mL/g DM | |||||||||
12 | Herb | 10.83 b | 9.40 b | 13.33 a | 9.49 b | 0.54 | 0.0029 | 0.9572 | 0.1585 |
Pine | 11.95 a | 8.68 b | 11.13 a | 8.36 b | 0.51 | 0.0024 | 0.0048 | 0.5125 | |
24 | Herb | 19.33 | 16.84 | 18.39 | 17.24 | 1.10 | 0.4242 | 0.3173 | 0.5270 |
Pine | 17.22 | 12.90 | 19.83 | 11.30 | 3.23 | 0.2955 | 0.4683 | 0.6308 | |
Carbon dioxide, mL/g DM | |||||||||
12 | Herb | 61.30 b | 64.26 b | 88.00 a | 64.66 b | 3.20 | 0.0012 | 0.0348 | 0.0116 |
Pine | 45.47 b | 41.36 bc | 52.44 a | 37.38 c | 2.06 | 0.0047 | 0.2098 | 0.0553 | |
24 | Herb | 89.69 | 93.75 | 99.07 | 94.21 | 4.37 | 0.5405 | 0.3307 | 0.4248 |
Pine | 56.73 | 50.79 | 57.75 | 48.01 | 4.72 | 0.4465 | 0.3809 | 0.7546 |
Incubation Time (h) | Treatment | Extract Concentration (1) (mg/L) | SEM (2) | p Value (3) | |||||
---|---|---|---|---|---|---|---|---|---|
0 | 30 | 50 | 70 | T | L | Q | |||
Total VFA, mM | |||||||||
12 | Herb (4) | 53.85 c | 54.74 b | 59.77 a | 54.79 b | 0.27 | <0.0001 | 0.0001 | <0.0001 |
Pine (5) | 53.98 | 60.36 | 59.37 | 55.70 | 1.95 | 0.1458 | 0.4766 | 0.0339 | |
24 | Herb | 62.40 | 63.15 | 65.17 | 62.46 | 1.50 | 0.5527 | 0.6974 | 0.3463 |
Pine | 61.52 | 60.88 | 64.91 | 62.49 | 1.22 | 0.1815 | 0.2500 | 0.7364 | |
Acetate, mM | |||||||||
12 | Herb | 36.58 b | 36.90 b | 40.80 a | 37.04 b | 0.28 | <0.0001 | 0.0019 | 0.0004 |
Pine | 36.20 | 39.68 | 39.22 | 36.40 | 1.79 | 0.4255 | 0.8482 | 0.1172 | |
24 | Herb | 42.62 | 42.91 | 44.49 | 42.60 | 0.91 | 0.4475 | 0.6737 | 0.3417 |
Pine | 40.90 | 40.99 | 42.46 | 41.48 | 0.93 | 0.6371 | 0.4530 | 0.7138 | |
Propionate, mM | |||||||||
12 | Herb | 10.56 b | 10.77 b | 11.88 a | 10.61 b | 0.13 | 0.0003 | 0.0367 | 0.0015 |
Pine | 10.93 b | 11.74 a | 11.92 a | 10.92 b | 0.17 | 0.0049 | 0.5144 | 0.0009 | |
24 | Herb | 12.59 | 12.64 | 13.28 | 12.47 | 0.36 | 0.4306 | 0.8262 | 0.3357 |
Pine | 12.52 b | 12.04 b | 13.44 a | 12.17 b | 0.28 | 0.0269 | 0.7993 | 0.3740 | |
Butyrate, mM | |||||||||
12 | Herb | 6.71 | 7.07 | 7.09 | 7.14 | 0.21 | 0.4762 | 0.1712 | 0.5345 |
Pine | 6.84 | 8.94 | 8.23 | 8.38 | 0.48 | 0.0750 | 0.0744 | 0.0800 | |
24 | Herb | 7.20 b | 7.59 a | 7.41 ab | 7.40 ab | 0.24 | 0.7228 | 0.6249 | 0.4097 |
Pine | 8.11 | 7.85 | 9.01 | 8.84 | 0.36 | 0.1424 | 0.0828 | 0.6160 |
Item | Extract Concentration (1) (mg/L) | SEM (2) | p Value (3) | |||||
---|---|---|---|---|---|---|---|---|
0 | 30 | 50 | 70 | T | L | Q | ||
Pinus densiflora (12 h) | ||||||||
Ruminococcus albus | 1.00 | 1.15 | 0.93 | 1.00 | 0.07 | 0.1930 | 0.5809 | 0.3892 |
Fibrobacter succinogenes | 1.00 a | 0.85 a | 0.27 b | 1.02 a | 0.09 | 0.0010 | 0.1474 | 0.0025 |
Ruminococcus flavefaciens | 1.00 b | 0.86 b | 1.46 a | 0.93 b | 0.07 | 0.0032 | 0.2266 | 0.0945 |
Ciliate associated methanogen | 1.00 b | 1.42 b | 3.06 a | 2.03 ab | 0.35 | 0.0165 | 0.0155 | 0.1927 |
Pinus densiflora (24 h) | ||||||||
Ruminococcus albus | 1.00 | 1.07 | 0.88 | 1.09 | 0.09 | 0.3989 | 0.8606 | 0.5979 |
Fibrobacter succinogenes | 1.00 b | 1.31 b | 0.45 c | 2.18 a | 0.17 | 0.0020 | 0.0143 | 0.0085 |
Ruminococcus flavefaciens | 1.00 | 0.93 | 1.36 | 1.11 | 0.11 | 0.1553 | 0.1643 | 0.7306 |
Ciliate associated methanogen | 1.00 | 0.83 | 1.16 | 1.22 | 0.23 | 0.6440 | 0.4032 | 0.5144 |
Mentha Canadensis (12 h) | ||||||||
Ruminococcus albus | 1.00 | 1.17 | 1.22 | 1.03 | 0.13 | 0.6194 | 0.7263 | 0.2397 |
Fibrobacter succinogenes | 1.00 a | 0.88 a | 0.17 b | 0.94 a | 0.06 | 0.0001 | 0.0061 | 0.0004 |
Ruminococcus flavefaciens | 1.00 | 1.02 | 1.35 | 0.95 | 0.14 | 0.2357 | 0.7238 | 0.2457 |
Ciliate associated methanogen | 1.00 b | 1.27 b | 3.39 a | 1.60 b | 0.26 | 0.0008 | 0.0083 | 0.0192 |
Mentha Canadensis (24 h) | ||||||||
Ruminococcus albus | 1.00 | 1.06 | 0.99 | 1.2 | 0.0766 | 0.2605 | 0.1699 | 0.3500 |
Fibrobacter succinogenes | 1.00 a | 0.93 a | 0.15 c | 0.49 b | 0.0691 | 0.0003 | 0.0003 | 0.2000 |
Ruminococcus flavefaciens | 1.00 | 1.16 | 2.39 | 1.34 | 0.202 | 0.0050 | 0.0313 | 0.0500 |
Ciliate associated methanogen | 1.00 b | 0.82 b | 1.12 a | 0.92 b | 0.2288 | 0.8152 | 0.9909 | 0.9500 |
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Lee, S.J.; Lee, Y.J.; Eom, J.S.; Kim, H.S.; Choi, Y.Y.; Jo, S.U.; Kang, S.N.; Park, H.Y.; Kim, D.H.; Lee, S.S. Effects of the Appropriate Addition of Antioxidants from Pinus densiflora and Mentha canadensis Extracts on Methane Emission and Rumen Fermentation. Animals 2020, 10, 1888. https://doi.org/10.3390/ani10101888
Lee SJ, Lee YJ, Eom JS, Kim HS, Choi YY, Jo SU, Kang SN, Park HY, Kim DH, Lee SS. Effects of the Appropriate Addition of Antioxidants from Pinus densiflora and Mentha canadensis Extracts on Methane Emission and Rumen Fermentation. Animals. 2020; 10(10):1888. https://doi.org/10.3390/ani10101888
Chicago/Turabian StyleLee, Shin Ja, Ye Jun Lee, Jun Sik Eom, Hyun Sang Kim, You Young Choi, Seong Uk Jo, Suk Nam Kang, Ha Young Park, Do Hyung Kim, and Sung Sill Lee. 2020. "Effects of the Appropriate Addition of Antioxidants from Pinus densiflora and Mentha canadensis Extracts on Methane Emission and Rumen Fermentation" Animals 10, no. 10: 1888. https://doi.org/10.3390/ani10101888