The bioactive activity of activated sludge was detected by the luciferase-ATP method.

: It is
that the treatment efficiency of the active sludge industry is closely related to the activity of
sludge
microorganisms. The greater the activity of the microorganism, the better the treatment effect; Typically, we measure microbial activity using indicators such as mixture suspension solid concentration (MLSS), mixed liquid volatile suspended solid concentration (MLVSS), sludge sedation ratio (SV), sludge volume index (SVI), sludge age (TS), etc.
With further research and advances in measurement techniques, it has been found that measuring the activity of microorganisms in activated sludge using MLSS (or MLVSS) is not accurate because MLSS (or MLVSS) also includes inactive substances unrelated to purification. That is, living cells cannot be separated from
organic
residues, such as ermic residues that have died, and it is not accurate to use it as a measure of biomass.
this, attempts have been made to reflect microbial activity using biological indicators such as
dehydrogenase
activity (DHA), ratio oxygen consumption rate (SOUR) and adenosine triphosphate (ATP). Among these indicators, ATP is widely found in biological cells, is an important product of energy metabolism in organisms and cell metabolism can use energy carriers, and participate in the organism
proteine
, fat
biochemical
synthesis, absorption and other reactions.
, by measuring the amount of ATP content, can directly reflect the cell activity, the number of microorganisms, and its measurement method is simple and fast. The application of ATP to the detection of biological activity of sludge has attracted wide attention.principle:
ATP content detection can be achieved by absorbent detection, light
detection or
isotope detection. The traditional light absorption detection method, low detection sensitivity, narrow linear range, it is difficult to accurately detect ATP conditions.
and the application of luciferase luminescence method to detect ATP, with very high sensitivity and very wide detection range, and simple, fast, is currently the most popular method. Based on firefly Luciferase catalytic luciferase catalytic luciferase oxidation, the method consumes ATP, emits an efficient luminous reaction of photons, and has the characteristics of very high luminous efficiency and a good linear relationship between luminous amount and ATP content.detection method:
1. Required equipmentspecial detectorspecial detection
reagents
box (including ATP
standard
, lysate, test fluid) active sludge sample
2. The test2.1 reagent preparation
(1) dilution of Lysis Buffer: dissolve Lysis Buffer at room temperature and dilute it with sterile water 5 times.
(2) the L/L test fluid: L/L Buffer room temperature dissolved, as a dilution of L/L substrate 50 times diluted, with L/L test fluid.
(3) ATP standards: 10x gradient dilution of ATP standards with Lysis Buffer to produce ATP standard samples with concentrations of 10-12 to 10-17mol/sl.2.2 ATP standard curve
(1) take 10-12 to 10-17mol/sl concentration of ATP standard samples each 10 μl, respectively, add 50 μl of L/L detection fluid, mixed into the light emitting detector test (light intensity RLU).
(2) to produce the ATP-RLU standard curve according to the ATP standard and the corresponding luminous intensity.2.3 Active Sludge Sample Detection
(1) extracts a certain amount of activated sludge and mixes it into a uniform suspension.
(2) Take 10 μl of activated sludge suspension, add 50μl Lysis Buffer, mix evenly and set aside for 5 min, then add 50μl L/L detection fluid and mix it evenly, put it into the tester to obtain the total ATP luminous strength.
(3) take 10 μl of activated sludge suspension, add 50 μl of pure water, mix evenly, set aside for 5 minutes, then add 50μl L/L detection fluid, and mix evenly, put into the detector test, can obtain free ATP luminous intensity in water. (Usually free ATP accounts for less than 1% of total ATP)
(4) brings the luminous intensity of the sample into the standard curve equation to obtain the ATP content in the sample.
(5) the active change of activated sludge can be obtained by detecting the activated sludge after different treatments or the activated sludge from different sources.
.