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The workflow for the experiment in the current study
Ingredient and nutrient composition of the experimental diet
Ingredients, % | Nutrients composition | ||
---|---|---|---|
Corn | 67.35 | Metabolizable energy, kcal/kg | 2,900 |
Soybean meal | 28.00 | Crude protein, % | 18.30 |
Soybean oil | 0.40 | Arginine, % | 1.14 |
Limestone | 1.90 | Lysine, % | 1.07 |
Monocalcium phosphate | 1.00 | Methionine, % | 0.47 |
Salt | 0.40 | Methionine + Cystine, % | 0.77 |
Lysine | 0.22 | Threonine, % | 0.73 |
Methionine | 0.17 | Tryptophan, % | 0.24 |
Choline chloride | 0.14 | Calcium, % | 1.01 |
Premix1 | 0.42 | Available phosphorus, % | 0.31 |
Total | 100 |
1Premix supplied per kilogram of diet: vitamin A, 13,200 IU; vitamin D3, 5,000 IU; vitamin E, 60 IU; vitamin K3, 8.0 mg; thiamine, 4.0 mg; riboflavin, 12.0 mg; pyridoxine, 12.0 mg; cobalamin, 0.4 mg; nicotinic acid, 80 mg; pantothenic acid, 24 mg; folic acid, 2.0 mg; biotin, 3.0 mg; iron, 70.0 mg; copper, 8.5 mg; zinc, 45.0 mg; manganese, 60.0 mg; iodine, 0.85 mg; selenium, 0.20 mg; cobalt, 0.25 mg
Example 3
Clostridium histolyticum ATCC 21000, strain 004 was inoculated into the starting culture with M #1 or M #2 and incubated at 37° C. for 16 hours. Ten milliliters of the starting culture (M #1 or M #2) and 10 mL Mg/vitamin solution (prepared separately by dissolving 8 g MgSO4, 1.2 g ferrous sulfate, 0.05 g riboflavin, 0.1 g Niacin, 0.1 g Calcium pantothenate, 0.1 g pimelic acid, 0.1 g pyridoxine, and 0.1 g thiamine in 1100 mL water, followed by sterilization by 0.22 μm filtration) was then transferred to each liter of M #3 or M #4 (or a variation thereof), and incubated for 22 hours. Clostridium histolyticum grew well with the OD600 reaching >2.5.
Example 3
Clostridium histolyticum ATCC 21000, strain 004 was inoculated into the starting culture with M #1 or M #2 and incubated at 37° C. for 16 hours. Ten milliliters of the starting culture (M #1 or M #2) and 10 mL Mg/vitamin solution (prepared separately by dissolving 8 g MgSO4, 1.2 g ferrous sulfate, 0.05 g riboflavin, 0.1 g Niacin, 0.1 g Calcium pantothenate, 0.1 g pimelic acid, 0.1 g pyridoxine, and 0.1 g thiamine in 1100 mL water, followed by sterilization by 0.22 pm filtration) was then transferred to each liter of M #3 or M #4 (or a variation thereof), and incubated for 22 hours. Clostridium histolyticum grew well with the OD600 reaching >2.5.
Composition and nutrient concentrations of basal diet (%, unless noted, as-is basis)
Item | Low phosphorus | Regular phosphorus |
---|---|---|
Ingredients | ||
Corn | 56.69 | 56.69 |
Soybean meal | 25.77 | 25.77 |
Distillers dried grains with solubles | 4.00 | 4.00 |
Calcium carbonate | 9.73 | 9.04 |
Dicalcium phosphate | - | 1.15 |
Soybean oil | 1.51 | 1.51 |
Sodium chloride | 0.26 | 0.26 |
DL-Methionine | 0.18 | 0.18 |
Choline chloride | 0.15 | 0.15 |
Montmorillonite | 0.71 | 0.25 |
Premix1 | 1 | 1 |
In total | 100.00 | 100.00 |
Nutrient levels | ||
Metabolizable energy, kcal/kg (calculated) | 2,600 | 2,600 |
Crude protein (calculated) | 16.5 | 16.5 |
Total phosphorus (calculated/analyzed) | 0.34/0.34 | 0.53/0.49 |
Non-phytate phosphorus (calculated) | 0.14 | 0.32 |
Calcium (calculated/analyzed) | 3.50/3.47 | 3.50/3.52 |
1Provided per kilogram of diet: manganese 60 mg, copper 8 mg, zinc 80 mg, iodine 0.35 mg, selenium 0.3 mg, vitamin A 8000 IU, vitamin E 30 mg, vitamin K3 1.5 mg, thiamine 4 mg, riboflavin 13 mg, pantothenic acid 15 mg, nicotinamide 20 mg, pyridoxine 6 mg, biotin 0.15 mg, folic acid 1.5 mg, and cobalamin 0.02 mg