A method for detecting sulfate-reducing bacteria in marine environments
A marine environment and sulfate technology, applied in the marine environment and marine corrosion field, can solve the problems of high cost and complicated testing process, and achieve the effect of low cost, simple equipment and high accuracy
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Embodiment 1
[0019] Centrifuge the test solution of sulfate-reducing bacteria at a speed of 6,000-80,000 rpm for 15-20 minutes, take the lower layer and place it in a selective medium (Postgete medium) for continuous culture, and the culture temperature Stable at 30-35°C. Take it out after 2-4 days of cultivation, take 2 ml of bacterial liquid and centrifuge it at a speed of 12000-14000 rpm for 15-20 minutes, and take the supernatant. Add oxidized glutathione solution to the supernatant, wherein the volume ratio of supernatant to oxidized glutathione is 9:1, the concentration of oxidized glutathione is 20mM, react at 37°C for 15-30 Minutes later, add a volume of 1-5 mg mL that is 1-1.2 times the supernatant to which oxidized glutathione was added -1 Papain solution, mix well and incubate at 37°C for 30-40 minutes. Then add 1% casein solution twice the volume of the solution to the solution (that is, add the supernatant of papain), react for 10-15 minutes, add trichloroacetic acid to term...
Embodiment 2
[0023] According to the method in Example 1, four kinds of bacteria (sulfate-reducing bacteria, Escherichia coli, Staphylococcus aureus and Vibrio alginolyticus) of the same concentration were treated according to Example 1 after being cultivated in Postgete medium for 4 days, By measuring the absorbance at 275nm of the hydrolyzate of thiol protease treated with various bacterial metabolites, the inhibition rate of the same concentration of different types of bacteria to thiol protease activity was calculated (see figure 2 ). The result shows that after deducting the background, the signal of the sulfate-reducing bacteria detected by the method provided by the invention far exceeds that of the other three bacteria, indicating that the detection method has good selectivity.
Embodiment 3
[0025] According to the method in embodiment 1, detect a series of different concentration sulfate-reducing bacteria (from 10 1 to 10 8 cfu / mL) after 2-4 days of culture, inhibition of thiol protease activity (see image 3 ). The results showed that when sulfate-reducing bacteria were cultured for 2 days, in 10 4 to 10 8 In the concentration range of cfu / mL, the inhibitory effect on thiol protease increased with the increase of the concentration of sulfate-reducing bacteria, and showed a good linear correlation. When the culture time was extended to 3 days, at 10 3 to 10 7 There is a good linear correlation within the concentration range of cfu / mL. After 4 days of culture, the detection range of sulfate-reducing bacteria can be reduced to 10 2 cfu / mL and maintain a good linear correlation.
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