Method and equipment for cultivating anaerobic ammonium-oxidizing bacteria
View Patent ↗The present invention can feed substrates without waste to generate seed sludge with high bacterial cell concentrations and can start up operation within a short time, in cultivating anaerobic ammonium oxidizing bacteria with ammonium and nitrite as substrates. Equipment for cultivating anaerobic ammonium-oxidizing bacteria, which cultivates, in a cultivation tank, novel anaerobic ammonium-oxidizing bacteria that anaerobically denitrify nitrite and ammonium used as substrates, comprises an ammonium feed device which feeds ammonium at a given concentration into the cultivation tank, a nitrite feed device which feeds nitrite at a given concentration into the cultivation tank, and a control device which controls a feed rate Y of the substrate.
1. A method for cultivating anaerobic ammonium-oxidizing bacteria capable of denitrifying ammonium and nitrite substrates, the method comprising:
cultivating the bacteria in a cultivation tank;
feeding a substrate into the cultivation tank; and
controlling a feed rate (Y) of the substrate after the bacteria enters a logarithmic growth phase to satisfy formula (I):
Y=A ×exp( K×T ) (I)
where Y is a feed rate of the substrate, A is a concentration of the substrate at the start of cultivation, K is a feed constant of the substrate, and T is a number of cultivation days after onset of the logarithmic growth phase.
2. The method of claim 1 , wherein:
the bacteria are bacteria having a DNA sequence of 16S rRNA gene whose particular region contains at least one sequence included in the group consisting of SEQ ID NO: 1-9.
3. The method of claim 1 or 2 , wherein:
an ammonium substrate and a nitrite substrate are fed into the cultivation tank; and
a ratio of ammonium nitrogen to nitrite nitrogen fed into the cultivation tank is controlled in a range of from about 1:0.5 to about 1:2.0.
4. The method of claim 1 or 2 , wherein the feed constant (K) of the substrate is set in a range of from about 0.05 to about 0.28.
5. The method of claim 3 , wherein the feed constant (K) of the substrate is set in a range of from about 0.05 to about 0.28.
6. The method of claim 1 or 2 , wherein the feed constant (K) of the substrate is set in a range of from about 0.06 to about 0.15.
7. The method of claim 3 , wherein the feed constant (K) of the substrate is set in a range of from about 0.06 to about 0.15.
8. The method of claim 1 or 2 , wherein the feed constant (K) of the substrate is set in a range of from about 0.07 to about 0.12.
9. The method of claim 3 , wherein the feed constant (K) of the substrate is set in a range of from about 0.07 to about 0.12.
10. The method of claim 1 or 2 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
11. The method of claim 3 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
12. The method of claim 4 further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
13. The method of claim 5 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
14. The method of claim 6 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
15. The method of claim 7 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
16. The method of claim 8 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
17. The method of claim 9 , further comprising:
increasing stepwise the feed rate (Y′) of the substrate to approach a rising curve represented by formula (I); and
controlling a nitrite nitrogen concentration in the cultivation tank so that the nitrite nitrogen concentration is 70 mg/L or less.
18. The method of claim 1 or 2 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
19. The method of claim 3 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
20. The method of claim 4 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
21. The method of claim 5 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
22. The method of claim 6 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
23. The method of claim 7 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
24. The method of claim 8 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
25. The method of claim 9 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
26. The method of claim 10 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
27. The method of claim 11 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
28. The method of claim 12 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
29. The method of claim 13 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
30. The method of claim 14 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
31. The method of claim 15 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
32. The method of claim 16 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).
33. The method of claim 17 , further comprising, after the bacteria enters the logarithmic growth phase:
determining an actual consumption rate (y) of the substrate by comparing a first concentration of the substrate fed into the cultivation tank and a second concentration of the substrate remaining in liquid discharged from the cultivation tank;
calculating a consumption constant (k) for the consumption rate of the substrate using formula (II):
y=a ×exp( k×T ) (II)
where y is the actual consumption rate of the substrate, a is a substrate concentration at the start of cultivating, k is the consumption constant of the substrate, T is the number of cultivation days from the onset of the logarithmic growth phase; and
setting the feed constant (K) to correspond to the calculated consumption constant (k).