Manipulation of Rice Straw Silage Fermentation with Different Types of Lactic Acid Bacteria Inoculant Affects Rumen Microbial Fermentation Characteristics and Methane Production
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolation, Identification and Characterization of LAB
2.2. Molecular Identification
2.3. Silage Preparation and Fermentation
2.4. Chemical Analyses and Fermentation Quality for Rice Straw Silage
2.5. In Vitro Rumen Fermentation and Digestibility
2.6. Quantification of Rumen Microbial Population by Real-Time PCR
2.7. Statistical Analyses
3. Results
3.1. Chemical Analyses and Fermentation Quality of Rice Straw Silage
3.2. In Vitro Rumen Fermentation Characteristics, Methane Production and DM Digestibility
3.3. In Vitro Rumen Microbial Populations
4. Discussion
4.1. Chemical Composition and Fermentation Characteristics
4.2. In Vitro Rumen Fermentation Characteristics, Methane Production and DM Digestibility
4.3. In Vitro Rumen Microbial Populations
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Ingredients (/kg DM) | g/kg DM |
---|---|
Alfalfa Hay | 314.1 |
Corn, grain | 170.0 |
Soybean meal | 133.0 |
Palm kernel cake | 251.1 |
Rice bran | 81.8 |
Sunflower oil | 20.0 |
Mineral premix | 5.0 |
Vitamin premix | 5.0 |
Ammonium chloride | 10.0 |
Limestone | 10.0 |
Chemical composition | g/kg DM |
DM | 850.2 |
CP | 208.3 |
EE | 52.5 |
NDF | 419.0 |
ADF | 253.0 |
Target Microorganism | Primer Sequences (5′–3′) |
---|---|
Fibrobacter succinogenes F | GGTATGGGATGAGCTTGC |
Fibrobacter succinogenes R | GCCTGCCCCTGAACTATC |
Ruminococcus albus F | CCCTAAAAGCAGTCTTAGTTCG |
Ruminococcus albus R | CCTCCTTGCGGTTAGAACA |
Ruminococcus flavefaciens F | CGAACGGAGATAATTTGAGTTTACTTAGG |
Ruminococcus flavefaciens R | CGGTCTCTGTATGTTATGAGGTATTACC |
General bacteria F | CGGCAACGAGCGCAACCC |
General bacteria R | CCATTGTAGCACGTGTGTAGCC |
General anaerobic fungi F | GAGGAAGTAAAAGTCGTAACAAGGTTTC |
General anaerobic fungi R | CAAATTCACAAAGGGTAGGATGATT |
Total protozoa F | GCTTTCGWTGGTAGTGTATT |
Total protozoa R | CTTGCCCTCYAATCGTWCT |
Total methanogens F | GCTCAGTAACACGTGG |
Total methanogens R | CGGTGTGTGCAAGGAG |
Total archaea F | ATTAGATACCCSBGTAGTCC |
Total archaea R | GCCATGCACCWCCTCT |
Treatments | Day | DM (%) | CP (%) | NDF (%) | ADF (%) | GE (Kcal) | Glucose 1 | Fructose 1 | Xylose 1 |
---|---|---|---|---|---|---|---|---|---|
Control | 15 | 34.8 a | 9.3 c | 69.2 a | 56.2 a | 15.7 | 5.3 a | 0.4 a | 0.4 |
30 | 33.6 a | 9.8 c | 70.2 a | 56.8 a | 15.9 | 4.8 b | 0.4 a | 0.4 | |
L. plantarum | 15 | 30.5 bc | 11.8 b | 66.8 c | 53.3 b | 15.5 | 3.5 c | 0.3 ab | 0.4 |
30 | 29.2 c | 12.7 a | 64.1 d | 49.4 c | 14.8 | 1.2 h | 0.1 c | 0.3 | |
L. salivarius | 15 | 33.2 a | 11.2 b | 68.9 a | 53.8 b | 15.7 | 3.6 c | 0.3 ab | 0.4 |
30 | 32.8 ab | 12.1 ab | 66.6 c | 51.9 c | 15.5 | 2.5 f | 0.2 b | 0.4 | |
L. reuteri | 15 | 32.5 ab | 10.9 bc | 67.6 b | 53.7 b | 15.9 | 3.2 d | 0.3 ab | 0.4 |
30 | 31.3 b | 10.9 bc | 66.6 c | 52.5 bc | 15.0 | 2.4 f | 0.2 b | 0.3 | |
L. brevis | 15 | 33.4 a | 10.6 bc | 66.9 c | 52.9 bc | 15.3 | 3.3 d | 0.3 ab | 0.4 |
30 | 32.4 ab | 10.9 bc | 66.6 c | 50.6 c | 15.1 | 3.0 e | 0.2 b | 0.3 | |
S. bovis | 15 | 32.6 ab | 12.4 a | 67.5 b | 52.1 bc | 15.7 | 3.4 cd | 0.3 ab | 0.3 |
30 | 31.5 b | 12.5 a | 64.8 d | 50.8 c | 15.3 | 1.9 g | 0.2 b | 0.3 | |
SEM 2 | - | 0.60 | 0.43 | 0.46 | 0.97 | 0.65 | 0.83 | 0.05 | 0.08 |
Treatments | Day | pH | Lactic Acid (g/Kg DM) | Acetic Acid (g/Kg DM) | Propionic Acid (g/Kg DM) | Butyric Acid (g/Kg DM) | NH3-N (%) | LAB (log cfu/g) |
---|---|---|---|---|---|---|---|---|
Control | 15 | 5.6 a | 5.1 f | 10.5 e | 1.3 b | 5.5 a | 0.05 | 5.2 c |
30 | 5.6 a | 9.4 e | 11.6 e | 1.6 b | 4.6 a | 0.05 | 5.8 c | |
L. plantarum | 15 | 5.2 ab | 19.6 c | 20.8 b | 1.9 ab | 1.7 cd | 0.04 | 6.6 bc |
30 | 4.4 b | 36.9 a | 24.1 a | 2.9 a | 1.7 cd | 0.05 | 8.8 a | |
L. salivarius | 15 | 5.4 a | 14.6 d | 13.4 d | 1.2 b | 3.3 b | 0.05 | 6.5 bc |
30 | 4.8 b | 22.4 bc | 19.1 bc | 1.3 b | 2.4 b | 0.05 | 7.3 b | |
L. reuteri | 15 | 5.5 a | 15.5 d | 14.2 d | 1.5 b | 2.2 c | 0.05 | 6.4 bc |
30 | 4.8 b | 26.6 b | 17.4 c | 2.1 ab | 2.1 c | 0.06 | 7.1 b | |
L. brevis | 15 | 5.3 a | 16.9 c | 14.4 d | 1.4 b | 2.6 b | 0.05 | 6.6 bc |
30 | 4.9 b | 24.1 b | 18.1 c | 1.7 b | 2.1 c | 0.05 | 7.3 b | |
S. bovis | 15 | 5.3 a | 19.9 c | 19.5 bc | 1.6 b | 1.7 cd | 0.04 | 6.8 bc |
30 | 4.3 b | 35.7 a | 22.5 ab | 2.5 a | 1.2 d | 0.05 | 8.2 ab | |
SEM 1 | - | 0.31 | 1.14 | 0.88 | 0.53 | 1.01 | 0.004 | 0.48 |
Treatments | Day | DMD | Total Gas 1 | pH | NH3-N | Total VFA (mM) | Acetic Acid | Propionic Acid | CH4 | CH4/Total Gas |
---|---|---|---|---|---|---|---|---|---|---|
Control | 15 | 21.4 f | 45.5 a | 6.9 | 14.3 | 65.6 g | 45.5 g | 13.6 a | 7.8 a | 0.17 a |
30 | 22.2 f | 45.0 a | 6.9 | 15.6 | 67.5 ef | 44.5 g | 12.7 ab | 7.9 a | 0.18 a | |
L. plantarum | 15 | 25.4 de | 42.0 bc | 6.9 | 15.3 | 74.7 c | 46.1 fg | 12.4 b | 4.2 c | 0.10 cd |
30 | 29.4 a | 37.5 c | 6.9 | 15.9 | 79.7 a | 54.4 a | 12.9 ab | 4.1 c | 0.11 cd | |
L. salivarius | 15 | 22.4 f | 43.5 b | 6.8 | 15.2 | 68.8 e | 47.0 f | 12.4 b | 6.1 b | 0.14 b |
30 | 26.4 cd | 41.0 c | 6.8 | 15.8 | 76.1 b | 49.7 d | 13.0 ab | 5.9 b | 0.14 b | |
L. reuteri | 15 | 22.4 f | 40.5 c | 6.8 | 14.9 | 70.6 d | 46.7 f | 13.3 a | 4.5 c | 0.11 cd |
30 | 26.4 cd | 40.0 cd | 6.8 | 15.5 | 74.5 c | 51.6 c | 13.2 a | 4.6 c | 0.12 c | |
L. brevis | 15 | 22.1 f | 43.0 b | 6.8 | 15.3 | 67.2 ef | 48.3 de | 12.5 b | 5.2 b | 0.12 c |
30 | 27.4 bc | 42.0 bc | 6.8 | 15.6 | 78.3 ab | 52.7 b | 13.6 a | 5.4 b | 0.13 bc | |
S. bovis | 15 | 24.4 ef | 40.5 c | 6.8 | 15.1 | 74.5 c | 49.2 d | 13.8 a | 5.7 b | 0.14 b |
30 | 28.4 ab | 39.5 cd | 6.8 | 15.7 | 78.9 a | 53.5 ab | 13.3 a | 5.2 b | 0.13 bc | |
SEM 2 | - | 0.68 | 0.84 | 0.03 | 0.48 | 1.07 | 0.48 | 0.32 | 0.26 | 0.004 |
Treatment 1 | Day | Fibrobacter succinogenes | Butyrivibrio fibrisolvens | Ruminococcus flavafaciences | Total Bacteria | Total Fungi | Total Protozoa | Total Methanogens | Total Archaea |
---|---|---|---|---|---|---|---|---|---|
Control | 15 | 1.54 b | 0.40 c | 1.20 c | 0.53 b | 0.76 b | 3.93 a | 7.62 a | 5.78 a |
30 | 1.61 b | 0.72 c | 1.31 bc | 1.05 ab | 0.78 b | 3.99 a | 7.57 a | 5.78 a | |
L. plantarum | 15 | 1.82 ab | 1.28 b | 1.91 b | 1.42 a | 1.07 ab | 3.62 ab | 7.34 ab | 5.34 ab |
30 | 2.50 a | 2.32 a | 2.68 a | 1.56 a | 1.37 a | 3.49 b | 7.11 b | 4.91 b | |
L. salivarius | 15 | 1.64 b | 1.12 c | 1.33 bc | 1.36 ab | 0.83 b | 3.84 a | 7.41 a | 5.50 a |
30 | 1.89 ab | 1.88 ab | 2.19 ab | 1.43 a | 1.15 a | 3.74 ab | 7.29 ab | 5.25 ab | |
L. reuteri | 15 | 1.64 b | 0.94 c | 1.31 bc | 1.33 ab | 0.83 b | 3.86 a | 7.47 a | 5.35 ab |
30 | 1.89 ab | 1.35 b | 2.07 ab | 1.42 a | 1.12 a | 3.66 ab | 7.36 ab | 5.01 b | |
L. brevis | 15 | 1.71 b | 1.13 c | 1.35 bc | 1.39 ab | 0.96 ab | 3.79 ab | 7.51 a | 5.43 ab |
30 | 2.12 a | 1.91 ab | 2.29 a | 1.44 a | 1.23 a | 3.66 ab | 7.43 a | 5.11 b | |
S. bovis | 15 | 1.77 b | 1.16 bc | 1.80 b | 1.42 a | 0.97 ab | 3.62 ab | 7.44 a | 5.22 ab |
30 | 2.24 a | 2.13 a | 2.55 a | 1.47 a | 1.34 a | 3.53 b | 7.22 b | 4.99 b | |
SEM 2 | - | 0.15 | 0.14 | 0.13 | 0.15 | 0.22 | 0.28 | 0.48 | 0.23 |
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Oskoueian, E.; Jahromi, M.F.; Jafari, S.; Shakeri, M.; Le, H.H.; Ebrahimi, M. Manipulation of Rice Straw Silage Fermentation with Different Types of Lactic Acid Bacteria Inoculant Affects Rumen Microbial Fermentation Characteristics and Methane Production. Vet. Sci. 2021, 8, 100. https://doi.org/10.3390/vetsci8060100
Oskoueian E, Jahromi MF, Jafari S, Shakeri M, Le HH, Ebrahimi M. Manipulation of Rice Straw Silage Fermentation with Different Types of Lactic Acid Bacteria Inoculant Affects Rumen Microbial Fermentation Characteristics and Methane Production. Veterinary Sciences. 2021; 8(6):100. https://doi.org/10.3390/vetsci8060100
Chicago/Turabian StyleOskoueian, Ehsan, Mohammad Faseleh Jahromi, Saeid Jafari, Majid Shakeri, Hieu Huu Le, and Mahdi Ebrahimi. 2021. "Manipulation of Rice Straw Silage Fermentation with Different Types of Lactic Acid Bacteria Inoculant Affects Rumen Microbial Fermentation Characteristics and Methane Production" Veterinary Sciences 8, no. 6: 100. https://doi.org/10.3390/vetsci8060100