Fermentation temperature | pH | DM (g/kg FM) | WSC | Total N | Lactic acid | Acetic acid | Ethanol | Propionic acid | n-Butyric acid | iso-Butyricacid (%) | NH3-N/total N | LAB1) | Clostridia spores |
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----------------------------------------- g/kg DM ------------------------------------------- | Log10 CFU/g FM | ||||||||||||
20°C | 3.74 | 299.4 | 35.94 | 13.78 | 59.24 | 80.01 | 10.66 | 1.17 | 0.53 | 0.28 | 5.41 | 7.85 | 2.80 |
10°C | 4.04 | 308.3 | 95.84 | 13.80 | 34.99 | 78.41 | 2.83 | 0.95 | 0.45 | 0.23 | 3.13 | 5.80 | 3.12 |
SEM | 0.056 | 2.639 | 11.386 | 0.102 | 5.582 | 0.637 | 1.590 | 0.282 | 0.053 | 0.088 | 0.442 | 0.389 | 0.154 |
p2) | *** | NS | *** | NS | ** | NS | *** | NS | NS | NS | *** | *** | NS |
DM, dry matter; FM, fresh materials; WSC, water-soluble carbohydrates; LAB, lactic acid bacteria; CFU, colony forming units; SEM, standard error of the mean; NS, not significant.
Storage temperature | Total N | DM (g/kg FM) | Lactic acid | Acetic acid | Ethanol | NH3-N/total N (% total-N) | Propionic acid | n-Butyric acid | iso-Butyric acid |
---|---|---|---|---|---|---|---|---|---|
Fermentation temperature (20°C) | |||||||||
5°C | 13.78 | 299.4 | 59.24 | 80.01ab 1) | 10.66b | 5.41a | 1.17 | 0.53 | 0.28 |
10°C | 13.87 | 307.9 | 55.92 | 70.64a | 7.50c | 5.35a | 0.94 | 1.32 | 0.07 |
15°C | 13.86 | 306.9 | 59.01 | 93.22bc | 8.10c | 5.42a | 0.90 | 1.64 | 0.10 |
20°C | 13.53 | 303.9 | 50.98 | 107.63c | 13.45a | 5.63ab | 0.76 | 1.02 | 0.13 |
25°C | 13.59 | 306.6 | 61.33 | 107.86c | 15.25a | 6.16b | 1.21 | 1.85 | 0.28 |
SEM | 0.080 | 1.25 | 1.912 | 4.081 | 0.895 | 0.086 | 0.144 | 0.208 | 0.045 |
Effects of increasing temperature | NS | NS | NS | *** | * | ** | NS | NS | NS |
Fermentation temperature (10°C) | |||||||||
5°C | 13.80 | 308.3 | 34.99 | 78.41A | 2.83 | 3.13A | 0.95 | 0.45 | 0.23 |
10°C | 13.63 | 314.0 | 40.49 | 80.96A | 2.79 | 3.40A | 1.10 | 0.22 | 0.19 |
15°C | 13.70 | 311.3 | 36.08 | 80.17A | 3.16 | 3.44A | 1.07 | 0.21 | 0.17 |
20°C | 13.68 | 304.8 | 44.39 | 81.16A | 2.52 | 3.55A | 1.41 | 0.54 | 0.36 |
25°C | 13.97 | 305.2 | 45.41 | 139.17B | 4.08 | 5.34B | 1.27 | 0.56 | 0.30 |
SEM | 0.089 | 1.60 | 1.531 | 5.569 | 0.213 | 0.182 | 0.054 | 0.059 | 0.039 |
Effects of increasing temperature | NS | NS | NS | *** | NS | *** | 0.063 | NS | NS |
Storage temperature | Culture media ((log10 CFU/g FM) | ||||
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LAB | Enterobacteria | Yeasts | Moulds | Clostridia spores | |
Fermentation temperature (20°C) 1) | |||||
5°C | 7.85a 2) | ND3) | ND | ND | 2.80 |
10°C | 7.82a | ND | ND | ND | 3.21 |
15°C | 7.67a | 4.02±0.07 | 3.93±0.08 | ND | 3.52 |
20°C | 7.81a | 4.71±0.47 | 4.85±1.21 | ND | 3.43 |
25°C | 8.19b | 4.78±0.10 | 4.44±1.11 | ND | 3.68 |
SEM d) | 0.049 | 0.517 | 0.504 | 0.108 | |
Effects of increasing temperature | **4) | NS | |||
Fermentation temperature (10°C) | |||||
15°C | 5.80A | ND | ND | ND | 3.12 |
10°C | 5.85A | ND | ND | ND | 3.37 |
15°C | 5.85A | ND | ND | ND | 3.28 |
20°C | 7.43B | ND | ND | ND | 3.54 |
25°C | 8.54C | ND | ND | ND | 3.58 |
SEM | 0.255 | 0.061 | |||
Effects of increasing temperature | *** | NS |
CFU, colony forming units; FM, fresh materials; LAB, lactic acid bacteria; SEM, standard error of the mean.
1) For each fermentation temperature (20°C and 10°C), values with different letters within the same column are statistically different (p≤0.05).
Number of clones sequenced | Most closely related bacterial sequence | GenBank accession No. of related sequence | Homology (%) | Environment from which related sequence was isolated |
---|---|---|---|---|
20E1) | Lactobacillus buchneri | JQ249065.1 | 100 | Fermented cucumber (USA) |
12C | Lactobacillus brevis | KC713915.1 | 100 | Fermented bamboo shoot (India) |
5D | Chryseobacterium sp. | AB461706.1 | 100 | Stems of field-grown soybeans (Japan) |
4A | Weissella koreensis | NR075058.1 | 100 | Kimchi (USA) |
1B | Leuconostoc citreum | KC417025.1 | 99 | Wheat flours (Italy) |
8 | Uncultured bacterium isolate | JX183833.1 | 98 | Jejunum, ileum and cecum of weaned piglets (China) |
6 | Pantoea agglomerans | KC355300.1 | 99 | Pepper (South Korea) |
2 | Lactobacillus oryzae | AB731661.1 | 99 | Fermented rice grain (Japan) |
2 | Pediococcus parvulus | AB601176.1 | 100 | Italian ryegrass silage (Japan) |
1 | Stenotrophomonas maltophilia | KC764984.1 | 100 | Tobacco rhizosphere soils (China) |
1) Clones labeled with letter “A” to “E” correspond to the marked out OTUs of Figure 3.
Number of clones sequenced | Most closely related fungal sequence | GenBank accession No. of related sequence | Homology (%) | Environment from which related sequence was isolated |
---|---|---|---|---|
23A1) | Candida humilis | AY493349.1 | 98 | Natural tequila fermentation |
8 | Uncultured Oidiodendron | JF796748.1 | 99 | Oil pumpkin flower (Austria) |
7 | Uncultured fungus clone | FJ757776.1 | 100 | Quercus macrocarpa phyllosphere (USA) |
6 | Uncultured Cladosporium | KC143740.1 | 100 | Human stool (France) |
6 | Uncultured Fusarium | HE977545.1 | 99 | Soil (UK) |
5B | Uncultured Davidiella | JX448366.1 | 100 | Agarwood (India) |
4 | Uncultured fungus clone | FJ757067.1 | 100 | Quercus macrocarpa phyllosphere (USA) |
3C | Uncultured Basidiomycota | HE977542.1 | 100 | Soil (UK) |
2 | Uncultured fungus clone | JN906946.1 | 100 | European beech (Fagus sylvatica) phyllosphere (France) |
2 | Uncultured fungus clone | FJ758346.1 | 100 | Quercus macrocarpa phyllosphere (USA) |
2 | Uncultured Alternaria | JQ346916.1 | 100 | Roots of herbs (endophyte) (China) |
1) Clones labeled with a letter “A” to “C” correspond to the marked out OTUs in Figure 3. Five other single uncultured fungus were also detected and cloning and/or sequence analysis of 7 fungus clones failed.