Table 7.
Relative abundance of bacterial taxa (phyla and genera level, sequenced targeting the V4 region of the 16s rRNA gene) associated rumen content (solid and liquid phase) collected at different sampling times (0, 6, and 12 h) from beef heifers fed a high forage diet (45% finely chopped grass hay and 45% barley silage) supplemented without 3-nitrooxypropanol (3-NOP, control) and with 3-NOP (150 mg/kg DM, 3-NOP)
| Item | Treatment (treat) | Sampling time (hour, H)4 | Rumen content (RC) | P-value3 | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Control | 3-NOP | SEM | 0 | 6 | 12 | SEM | Liquid | Solid | SEM | Treat | H | RC | |
| Observed amplicon sequence variants (ASV) | 695 | 611 | 17.4 | 675a | 654ab | 630b | 15.8 | 633 | 673 | 12.9 | <0.01 | 0.02 | <0.01 |
| Chao1 value | 696 | 612 | 17.5 | 676a | 655ab | 630b | 15.9 | 634 | 674 | 13.0 | <0.01 | 0.02 | <0.01 |
| Shannon index | 5.56 | 5.32 | 0.089 | 5.47 | 5.44 | 5.41 | 0.029 | 5.36 | 5.52 | 0.024 | 0.04 | 0.09 | <0.001 |
| Relative abundances1, % | |||||||||||||
| P Bacteroidates | 50.8 | 50.9 | 3.31 | 49.2 | 50.4 | 53.0 | 1.65 | 58.4 | 43.3 | 1.51 | 0.98 | 0.07 | <0.001 |
| g Prevotella | 39.0 | 35.3 | 3.20 | 34.5a | 37.3ab | 39.5b | 1.45 | 43.7 | 30.5 | 1.18 | 0.32 | <0.01 | <0.001 |
| g Rikenellaceae RC9 gut group | 3.83 | 5.05 | 0.290 | 5.35a | 4.32ab | 3.65b | 0.390 | 4.24 | 4.64 | 0.318 | 0.01 | <0.001 | 0.20 |
| g Prevotellaceae UCG-001 | 3.12 | 3.25 | 1.001 | 3.50 | 2.61 | 3.46 | 0.482 | 3.35 | 3.03 | 0.394 | 0.90 | 0.12 | 0.42 |
| g Prevotellaceae UCG-003 | 1.08 | 1.68 | 0.355 | 1.11 | 1.52 | 1.50 | 0.360 | 1.89 | 0.87 | 0.296 | 0.12 | 0.43 | <0.01 |
| g Prevotellaceae NK3B31 group | 0.78 | 0.96 | 0.346 | 1.18 | 0.70 | 0.73 | 0.285 | 0.79 | 0.95 | 0.227 | 0.62 | 0.16 | 0.48 |
| g Prevotellaceae UCG-004 | 1.14 | 2.13 | 0.425 | 1.14 | 1.64 | 2.12 | 0.345 | 1.62 | 1.65 | 0.260 | 0.06 | 0.02 | 0.92 |
| g Bacteroidales BS11 gut group | 0.33 | 0.37 | 0.074 | 0.41 | 0.34 | 0.31 | 0.097 | 0.33 | 0.38 | 0.074 | 0.57 | 0.47 | 0.92 |
| g Bacteroidates RF16 group | 1.27 | 1.47 | 0.180 | 0.97b | 1.45a | 1.68a | 0.176 | 1.58 | 1.16 | 0.141 | 0.32 | <0.01 | <0.01 |
| g Muribaculaceae | 0.53 | 0.63 | 0.102 | 0.76a | 0.27b | 0.70a | 0.141 | 0.62 | 0.54 | 0.093 | 0.40 | <0.01 | 0.40 |
| g Paraprevotella | 1.04 | 1.85 | 1.424 | 1.09 | 1.33 | 1.91 | 0.863 | 2.28 | 0.60 | 0.681 | 0.58 | 0.64 | 0.02 |
| g Uncultured | 0.73 | 0.86 | 0.070 | 0.86 | 0.76 | 0.77 | 0.111 | 0.86 | 0.74 | 0.089 | 0.11 | 0.62 | 0.17 |
| P Firmicutes | 32.6 | 31.1 | 1.49 | 33.1a | 32.9a | 29.6b | 1.20 | 27.6 | 36.1 | 0.98 | 0.36 | 0.01 | <0.001 |
| g Succiniclasticum | 15.6 | 18.0 | 1.34 | 17.9 | 16.6 | 15.8 | 1.13 | 16.0 | 17.6 | 0.92 | 0.10 | 0.18 | 0.10 |
| g Christensenellaceae R-7 group | 2.98 | 2.67 | 0.303 | 3.11 | 2.81 | 2.56 | 0.282 | 2.12 | 3.53 | 0.228 | 0.34 | 0.16 | <0.001 |
| g Ruminococcaceae | 0.61 | 0.55 | 0.159 | 0.62 | 0.60 | 0.53 | 0.137 | 0.40 | 0.76 | 0.105 | 0.69 | 0.77 | <0.01 |
| g Lachnospiraceae NK3A20 group | 1.60 | 1.80 | 0.284 | 1.60 | 1.94 | 1.52 | 0.307 | 1.14 | 2.26 | 0.250 | 0.50 | 0.38 | <0.001 |
| g Saccharofermentans | 1.34 | 0.79 | 0.240 | 1.44a | 1.09a | 0.67b | 0.146 | 0.75 | 1.38 | 0.116 | 0.04 | <0.001 | <0.001 |
| g uncultured f Ruminococcaceae | 2.14 | 0.26 | 0.636 | 0.88 | 1.44 | 1.27 | 0.685 | 1.20 | 1.19 | 0.484 | 0.02 | 0.60 | 0.98 |
| g Lachnospiraceae XPB1014 group | 1.29 | 0.67 | 0.246 | 0.90 | 1.19 | 0.84 | 0.201 | 0.65 | 1.30 | 0.160 | 0.06 | 0.16 | <0.001 |
| g Lachnospiraceae AC2044 group | 0.81 | 0.52 | 0.115 | 0.77 | 0.70 | 0.51 | 0.132 | 0.41 | 0.91 | 0.108 | 0.04 | 0.14 | <0.001 |
| g NK4A214 group f Oscillospiraceae | 1.50 | 1.04 | 0.163 | 1.35 | 1.39 | 1.07 | 0.194 | 1.11 | 1.43 | 0.158 | 0.02 | 0.21 | 0.05 |
| g Pseudobutyrivibrio | 0.94 | 0.78 | 0.135 | 1.02 | 0.79 | 0.77 | 0.126 | 0.47 | 1.25 | 0.098 | 0.29 | 0.09 | <0.001 |
| g Veillonellaceae UCG-001 | 0.79 | 0.71 | 0.093 | 0.86a | 0.80a | 0.60b | 0.093 | 0.71 | 0.80 | 0.075 | 0.46 | 0.02 | 0.26 |
| g Mogibacterium | 1.08 | 0.58 | 0.066 | 0.70 | 0.95 | 0.84 | 0.098 | 0.58 | 1.08 | 0.082 | 0.01 | 0.05 | <0.001 |
| P Proteobacteria | 2.42 | 2.83 | 0.963 | 0.32c | 2.42b | 5.13a | 0.671 | 3.67 | 1.57 | 0.533 | 0.68 | <0.001 | <0.001 |
| P Fibrobacteres | 7.55 | 8.73 | 1.852 | 10.54a | 7.97b | 5.91b | 1.260 | 4.81 | 11.5 | 1.02 | 0.54 | <0.01 | <0.001 |
| P Spirochaetae | 4.82 | 4.70 | 1.667 | 3.68b | 4.66b | 5.95a | 0.517 | 3.81 | 5.72 | 0.422 | 0.95 | <0.001 | <0.001 |
| g Treponema | 4.59 | 4.60 | 1.593 | 3.45b | 4.50ab | 5.82a | 0.524 | 3.65 | 5.54 | 0.427 | 0.99 | <0.001 | <0.001 |
| P Verrucomicrobia | 0.95 | 0.68 | 0.170 | 1.14a | 0.71b | 0.59b | 0.165 | 1.04 | 0.59 | 0.133 | 0.15 | <0.01 | 0.001 |
| g WCHB1 41 | 0.90 | 0.53 | 0.087 | 1.03a | 0.60b | 0.52b | 0.140 | 0.90 | 0.54 | 0.110 | 0.01 | 0.001 | 0.002 |
| P Planctomycetota | 0.43 | 0.49 | 0.111 | 0.65a | 0.41b | 0.38b | 0.118 | 0.42 | 0.54 | 0.107 | 0.97 | 0.04 | 0.26 |
| P Patescibacteria | 1.67 | 1.39 | 0.692 | 2.08a | 1.26b | 1.25b | 0.297 | 1.74 | 1.32 | 0.232 | 0.69 | <0.01 | 0.08 |
| Protozoa, 105 cell/mL2 | 4.71 | 5.73 | 0.612 | 6.52a | 4.65b | 4.53b | 0.314 | na | na | na | 0.14 | <0.001 | na |
1 p = phylum, g = genus, f = family.
2Protozoa were identified and counted under microscope following Ogimoto and Imai (1981). Identified ruminal protozoa genera included: Isotricha spp., Dasytricha spp., Entodinium spp., Diplodinium spp., Ostracodinium spp., Metadinium spp., Polyplastron spp., and Osphyrscolex spp.
3Significance was declared at P ≤ 0.05 and a tendency to significance was declared at 0.05 < P ≤ 0.10.
4Mean values with a different superscripted letter differ significantly among sampling times.