SU1684664A1 - Method of biological testing of industrial sewage toxicity - Google Patents

Description

The invention relates to the biological purification of industrial wastewater, can be used to determine the toxicity of wastewater of individual industries, individual components and multicomponent systems of contaminants when taken for biological treatment, and discharged into reservoirs, as well as for operational control of biological treatment facilities.

The purpose of the invention is to increase the accuracy and sensitivity of the biotesting method of toxicity of industrial wastewaters and their components by establishing a violation of the ability of the debris of microorganisms of the active silt ecosystem

specific dye under the action of toxic wastewater and their components.

The method is carried out as follows. In flasks placed 200 ml of sludge suspension of existing treatment facilities. One flask was left as a control, different amounts of wastewater (10-50 ml, depending on the probable toxicity) or components of the wastewater in different concentrations were added to the other (s). The flasks are placed on magnetic stirrers and, stirring, are incubated for 1 hour at 37 ° C. The duration and temperature of incubation are determined by the optimization of the experimental conditions. At the end of the incubation of

O5

wasp

4b

s & o: s

control and prototypes take 10 ml of sludge suspension, washed three times with phosphate buffer solution pI 7.0. centrifuged at, oooo rpm 10 min. The precipitate is transferred to a mechanical disintegrator (glass mortar and pestle on thin sections), disintegration is carried out for 5 min at k ° C. The disintegratable sludge mixture is transferred into a flask, 5 ml of phosphate buffer pH 7.0 and Triton X-100 20 mg / ml in final concentration are added. The flasks are placed on magnetic stirrers and continue to disintegrate c) 5 for 2 hours at 37 ° C. Disintegrate is centrifuged at O JOO rpm for PM - 10 minutes, thus supernatant containing proteins of microorganisms of the activated sludge ecosystem is obtained. Then in the supernatant the total protein is determined according to Biuret or Lowry and the LRNA fraction of methods of electrophoresis in flat blocks of polyacrylamide gel. For the analyzed samples, 25 volumes of 59 µl mixed with 0% rastofo, sucrose was applied to the star- line. water leads to a change in the electrophoretic mobility of the protein fractions compared to the control, with the fractions being detected in horseshoes with teeth on the edges. Such qualitative changes in protein fractions indicate the onset of the toxic effect of wastewater and its components on microorganisms of the activated sludge ecosystem. An increase in the concentration of toxic components of wastewater leads to a gradual increase in the violation of the fixation of a specific dye by proteins up to the complete disappearance of the colored fractions from the floor of electrophoregram. Moreover, the total amount of protein in the samples and in the control and in the experiments remains approximately at the same level.

Example 1. In k flasks placed 2UO ml sludge suspension 1-st stage of biological wastewater treatment. One flask was left as a control; 50 mg / g sludge, 100 mg / g sludge and 150 mg / g sludge of Progress detergent were added to the others. To the specified detergent active pH pH j, 2. Electrophoresis is carried out at a zone sludge adapted. Flasks are placed on

current strength 5.0 mA / cm for the first 30 minutes, then 10.0 mA / cm until the end of electrophoresis. The obtained protein fractions are stained and fixed, for which purpose electrophoregrams are placed in a 1% solution of black acid in 71 ° acetic acid for 1 J min. Removal of excess dye is carried out by soaking the gel plates in an acetic acid solution. To accelerate the background fading, the acetic acid solution is changed k times a day.

Express staining and fixation of electrophoregrams are carried out in the following way: after electrophoresis, the cathode chamber of the device is filled with a 1% solution of black amido in 1% acetic acid and the process continues for 1C rank. After staining the gels, rsop dye is replaced with acetic solution acids and continue electrophoresis until background staining is completely removed.

For a qualitative assessment of the effect of toxic wastewater and its components on proteins of microorganisms of activated sludge, it is enough to visualize electropherograms without densitoglu, h LI- ti roognip

, y) 5 20 25

66) 4

The effect of minimal toxic concentrations of wastewater components leads to a change in the electrophoretic mobility of the protein fractions compared to the control, with the fractions occurring in the horseshoe c with teeth on the edges. Such qualitative changes in protein fractions indicate the onset of the toxic effect of wastewater and its components on microorganisms of the activated sludge ecosystem. An increase in the concentration of toxic components of wastewater leads to a gradual increase in the violation of the fixation of a specific dye by proteins up to the complete disappearance of the colored fractions from the floor of electrophoregram. Moreover, the total amount of protein in the samples and in the control and in the experiments remains approximately at the same level.

Example 1. In k flasks placed 2UO ml sludge suspension 1-st stage of biological wastewater treatment. One flask was left as a control; 50 mg / g sludge, 100 mg / g sludge and 150 mg / g sludge of Progress detergent were added to the others. To the specified detergent active5

0

Q

five

five

Magnetic stirrers and with stirring, incubate for 1 h at 3V ° C0. Sampling, electrophoresis and detection of protein fractions are carried out as described above. Experimental data on the known and proposed methods are presented in table. 1 and 2 respectively.

From the data obtained in a known manner, it can be seen that, under the action of CMC Progress at a concentration of 50 mg / g sludge, a decrease in the total activity of dehydrogenases occurs at 1%, 5% of the control, which indicates the absence of toxic effects on microorganisms of activated sludge. From the data of the proposed method, it is evident that with the action of CMC Progress at a concentration of 50 mg / g sludge, 3 protein fractions are detected, however, they differ qualitatively from the electrophoregram fractions of control samples by the appearance of teeth at the edges of the fractions, changes in shape and electrophoretic mobility. These signs are characteristic of the toxic effect of CMC of such concentration on microorganisms of the activated sludge ecosystem. The action of a concentration of 100 mg / g of sludge leads to the departure of one fraction from the floor of electrophoregram, and the action of 150 mg / g of sludge leads to the disappearance of all three fractions of protein from the floor of electrophoregrams. The amount of total protein according to Lowry remains approximately at the same level in all series of experiments both in the control and in the experiment.

The data from this series of experiments suggests a higher sensitivity of the proposed method compared to the known one.

PRI me R 2.

In 2 flasks are placed 200 ml of silt suspension of the 1st stage of biological wastewater treatment. One flask was left as a control, and another 120 mg / g of ammonia sludge was added to the other, the salvo emissions of which are characteristic of the production. The flasks are placed on magnetic stirrers and incubated for 1 hour at 37 ° C with stirring. Sampling, electrophoresis and detection of protein fractions are carried out in the same way. Experimental data on the known and proposed methods are presented in Table. 2 and 3 respectively.

Experimental data obtained in a known manner, indicate a pronounced toxicity of ammonia in such a concentration relative to sludge microorganisms (a decrease in the initial activity of dehydrogenases by 0.1%). From the data of the proposed method, it is clear that instantaneous release of ammonia at such a concentration leads to the disappearance of all three fractions from the floor of electrophoregram, which indicates acute toxicity for the activated sludge ecosystem. This series proves that the proposed method is much more sensitive than the known one.

Example-B2 flasks are placed in 200 ml sludge suspension of the 1st stage of biological wastewater treatment. One flask is left as a control, to the other is added 150 mg / g sludge of normal paraffins, also characteristic of the wastewater of the indicated production. Flasks are placed on magnetic

Mixers and with stirring, incubate for 1 hour at 37 ° C. Samples are obtained, electrophoresis and protein fractions are detected in

the same sequence as before

Q

5 0 5

0

five

examples. These on experiments are presented in Table. 5 and 6, respectively.

From the data table. It can be seen that, according to a known method, n-pair-tphins in the concentration studied are not toxic with respect to microorganisms of activated sludge. The series carried out by the proposed method shows that of the three fractions detected in the control samples, under the action of n-paraffins, only one protein fraction with teeth on the edges is detected, which indicates a pronounced toxic effect of this component on activated sludge. This once again confirms the great feeling about the stabilities of the proposed method.

The use of the proposed method of biotesting the toxicity of industrial wastewater will make it possible to establish with greater sensitivity the minimum concentrations of wastewater components, at which toxic effect on microorganisms of the ecosystem of raw sludge begins, and the concentrations that act on activated sludge are acutely toxic.

The use of the method at biological wastewater treatment plants of industrial plants, urban aeration stations, industrial research institutes and biomonitoring services will ensure the sanitary effect through operational monitoring of the state of wastewater when receiving biological treatment and discharge of water bodies, will make more efficient wastewater management .

40

Claims (1)

Formula invented and five 0 The method of biotesting toxicity of industrial wastewater, involving the sampling of microorganisms of the activated sludge ecosystem, their incubation with wastewater components of various concentrations, subsequent centrifugation, staining of protein samples, and classification of the studied components of the wastewater to toxic ones by changing the staining intensity prototypes, characterized by the fact that, in order to improve the biotesting accuracy, after incubation, the activated sludge microorganisms are disintegrated ova- tion, and electrophoresis of the obtained protein samples in flat blocks of polyacrylamide gel with subsequent staining of the cytophoretic mobility of the fractions protein fractions specific., protein and violation of fixation by the dye, and the intensity of the staining: those: l protein fractions in the experiment are compared by shape change; elenenii with control. Table 1 Table 3 The known method Total activity of activated sludge dehydrogenase, Mr formazan / g sludge Control 2.67. + M1 Decrease %% 3 3 3 3 3 Experience (120 mg ammonia / g sludge) 1.59 iO, 08 - / 40.1 Table k 8.1 8.0 9, 7.3 7.6 9168 66 J Table 5 The total activity of activated sludge dehydrogenase, mg form per / g Control Experience (150 mg of n-paraffins / g sludge) 2, 56 iO., T Reduction,, 0 Table Kontpaypyt (n-paraffin, 150 mg / g Total Bees Fractions Total Protein Fractions b lok for protein, for Lowry, (amount Lowry count wmg / ml mg / ml 8.537.91 7,737,61 10,239.11 7,038,01 8,238,21