Method of estimating condition of water source in an agricultural enterprise effluent zone

FIELD: environmental monitoring.

SUBSTANCE: invention relates to hygiene and sanitary of freshwater reservoirs and is meant to be used for microbiological testing of condition of water source in an agricultural enterprise effluent zone, in particular in the effluent zone of poultry factories and pig-breeding farms. For this aim, water is sampled at least in two zones: in the effluent zone of agricultural enterprise and in the zone, where influence of agricultural enterprise effluent over different periods is excluded. Then, contents of ammonia compounds and enzymatic activities of urease-carbamidamidohydrolase in samples are measured. Obtained data are processed: contents of ammonia compounds and enzymatic activities in different samples are compared to each other and dynamic of changes in data in samples taken from different zones are compared.

EFFECT: enabled quick testing at high accuracy in estimations and minimized labor and means involved.

2 tbl

 

The invention relates to hygiene and sanitation freshwater and is intended for carrying out of sanitary-hygienic and microbiological testing of water sources in the area of sewage farms, in particular poultry and pig farms, industrial encompassed in nearby ponds.

Known “Method of determining the toxicity of natural and waste waters” (see A.S. No. 1194877, 12 Q 1/00, publ. 30.11.85, bull. No. 44), in which the quality of water is determined additionally by contacting water with the test organisms, which use the flagellate of the genus Chlamudomonos, and on the toxicity of water is judged visually by the disappearance or by the presence of bands accumulation of parasites in the control tubes.

The disadvantages of this method of testing is it some complexity, as is required in advance to prepare and keep spetsialnye test organisms and to have a checklist on these test organisms.

Also known “Method of determining the degree of saprobity water freshwater (see RF Patent №2123533, 12 Q 1/04, publ. 20.12.98, bull. No. 35), which conducts microbiological isolation of microalgae, and the selected algae define lisicina and antilysocyme activity, then determine the index of saprobity each species, calculate the average value and its value with the field of the degree of saprobity of the reservoir and the water quality.

With good reliability of the results by this method the disadvantages should be considered a large amount and duration necessary research and calculations and, in addition, such work requires highly qualified professionals to determine the tested species allocated microalgae and sophisticated equipment for testing.

The objective of the invention is to develop additional assessment of the water source, particularly in the area of discharge of industrial wastewater agricultural enterprises, in which it would be possible to test the quality of wastewater in the existing chemical laboratories, using average technicians on the usual laboratory equipment and in obtaining rapid results test analyses for high reliability assessment with a minimum expenditure of labor and resources to the analysis.

The task is solved in that the test analyses further define the activity of the enzyme urease in the study and control water samples taken simultaneously from the contaminated source and relatively clean in the same zone, at a depth of 0.3 to 1 m at a distance of 1-2 m from the edge of the shore, in particular urease activity and ammonia in the samples, and then determine their attitude, and on water quality and the condition of the pond judged by this dormancy is the result, and the point of taking control (relatively pure) sample water feature outside of the zone of significant pollution of agricultural enterprises, at a distance of 1-2 km upstream from the confluence of runoff agricultural enterprises. The content of ammonium compounds in water samples to determine the photocolorimeter in parallel with the determination of the activity of the enzyme urease-carbamidomethylated.

It is known that one of the main pollutants in the runoff of agricultural enterprises is urea. Urea gets into the water in the composition of plant residues, manure and various other wastes of livestock and poultry. It is formed in the water as the products of transformation of nitrogenous organic compounds, proteins and nucleic acids.

The proposed method is based on the ability of aquatic ecosystems to cleanse itself from contaminants of agricultural origin by transformation of the nitrogen polluting substances (urea, and others) with the participation of the enzyme urease secreted aquatic biota (all living components of aquatic ecosystems).

Determination of the activity of one of the most important enzymes involved in this process (urease-carbamidomethylated CFS.) serves as a test for assessment of surface water sources in the area of agricultural use of the project, i.e. a measure of the degree of degradation of the studied aquatic ecosystems, as well as for environmental regulation and the determination of the maximum allowable anthropogenic impacts on aquatic biota.

High content of urea in water runoff is the level of activity of the enzyme urease, which catalyzes the hydrolysis of urea. The end product of hydrolysis are ammonia and carbon dioxide. Therefore, by controlling the level of activity of the enzyme urease and ammonia and determining their ratio in the samples and in the control of water taken, it is possible to assess the condition of the water source in the area of agricultural runoff. The benchmark adopted the ratio of the potential activity of the enzyme urease is the ratio of the rate of activity of urease (mg NH3mg/l) ammonia in samples (mg NH3mg/l).

This relative indicator characterizes the ability of aquatic ecosystems to self-purification from organic ingredients, due to the intensity of the process of ammonification agricultural waste, which is expressed by the equation (see workshop on the chemistry edited Vdenisa, M., Ed. Moscow state University, 1989. pp.92-94).

urease, N2About

The method is as follows.

During the examination on the object outline the place of sampling. The first point is at the usually scheduled in place of sewage from a contaminant at a distance of 1-2 meters from the shore at a depth of 0.3 to 1 meter, the following points at distances of 50-60 meters from one another downstream, and two of them on the opposite side from the place of sewage.

A distance of 50-60 meters adopted by the results of practical experiments as providing the required accuracy of the forecast and does not require a lot of navigation.

Simultaneously with the sampling of water in the target area take control water samples at a distance of 1-2 km upstream, where any influence from the polluter. The volume of water samples for analyses adopted 2-2,5 liters. Samples are packaged and transported to the laboratory for analysis immediately after delivery, the activity of the enzyme urease determine calorimetric methods using device type FEC - 56M and the ammonia determined in parallel by the same method (photocalorimetric) on the same device.

To determine the colorimetric method of urease activity using the following reagents:

1. 1 N. KCl;

2. Phosphate buffer pH 6,7;

3. The Nessler reagent;,

4. 50% solution of potassium-sodium tartrate.

The course of the analysis. In the test tube is placed 1 ml of the investigated water, add 2 ml of phosphate buffer (pH 6,7), 2 ml of 10% urea solution. The tube close glass stopper, shake and put in a thermostat at 37 t With° for 24 hours. After incubation, add in a test tube, 3 ml of 1N. rest the RA KCl, shake for 5 minutes to displace the water of ammonia, filter, placed in a flask of 50 ml, add 30 ml of distilled water. To mix. Add 2 ml of Nessler reagent, stir and bring to the mark with water. To colorimetrically on the FEC /-490 nm/.

The amount of ammonia is calculated on pre-prepared calibration curve.

Standard solution: 0.1 mg NH3in 1 ml of water. Control - boiled water (without substrate).

Urease activity was expressed in mg NH3to 1 liter of water per day. Calculation of NH3on the calibration line.

where a is the amount of ammonia according to the schedule;

b - amount of ammonia in control;

n - quantity of water taken for research.

R - breeding;

1000 - conversion to 1 litre

The obtained measurement results are processed by methods of mathematical statistics bring to the table, build charts and analyzing and comparing the results obtained on the test points for taking water samples, make conclusions about the status of the test water and the influence of industrial agricultural runoff and outline measures for the protection of the purity of the gutters.

Example No. 1. Assessment of water and efficiency of water protection measures to reduce biological contamination and toxicity of runoff from landfills on which I store bird droppings on the Sverdlovsk poultry.

The studies were conducted in the waters of the river “Source” of the tributary of the river Iset flowing on the territory of Sverdlovsk region, near the southern outskirts of the city, which is constantly coming (reset) the effluent from the Sverdlovsk poultry.

Sample No. 1 (control) was taken at a distance of 3 km upstream to the confluence drains, sample No. 2 - 2 km after their confluence, also took a sample of water in the drain the swamp is a natural filter, not contaminated poultry waste (sample No. 3)drain the swamp - waste poultry (sample No. 4), and the overall flow of the two wetlands at a distance of 20 metres from the junction between them (sample No. 5).

Samples of water were taken according to the above rules, at a distance of 1-2 meters from the shore, depth of water (0.3 to 1 m), in 3 replications. Sampling was conducted during the summer and autumn of 2000.

Treatment taken during the analyzed period, water samples by the proposed method estimates and standard techniques were performed in the laboratory of the Department of ecology and zoohygiene Ural State agricultural Academy. The results of studies after mathematical processing are summarized in table No. 1 and No. 2.

1 study to assess the condition of the water source by determining the activity of urease water samples with different degree of pollution by sewage poultry paraslisante the enzyme urease activity and ammonia in waters runoff from the bog - landfill (samples 4, 5) 22.7-115% compared to the clean waters of the swamp (sample 3) during the summer period.

1K
Table 1.
The dynamics of the enzyme urease activity in the research process (in different periods).
Period№p/pSod s ammonium Conn-s in water (NH3mg/l)Urease activity (NH3mg/l)The ratio of the potential activity of the enzyme (CPU)
9.06.2000 St1K13,1±0,3193±9,0130
I23,0±0,2990±8,8830
start38,0±0,74110±of 10.2117
summer428,0±2,17138±13,24a 4.9
 527,0±2,07134±13,14a 4.9
23.06.20001K2,8±0,26105±9,3133
II23,0±0,27119±10,3440
the middle3,7± 0,54135,7±12,3420
summer428±2,14300,0±27,3410,9
 527±2,13273,0±26,91the 10.1
15.09.20001K5,0±0,35107,2±9,3121
III24,8±0,45to 102.3±9,7121
start320,0±1,97120±11,346
autumn428,0±2,73135±12,714,8
 522,5±2,17116,5±11,325,2
29.09.20001K10,0±0,97122,0±10,3412,1
IV215,0±1,34148,5±14,3310,0
the middle315,0±1.27148,0±14,8710,0
autumn422,5±2,33158,0±13,937,0
 522,5±2,21160,0±1%.347,1
11.10.20003,0±0,2985,5±8,3128,5
V26,3±0,6171,3±7,1311,1
end310,0±0,9452,5±5,145,2
autumn418,3±1,3775,8±7,314,1
 520,0±1,5880,0±to $ 7.914,0
1to CONTROL

56 -
Table 2.
The research results of the samples of effluent under the existing method.
The study periodsNo. of samples (K)Ammonium Conn. mg/lThe oxidation mg/lThe number of microbes in 1 ml of water thousand unitsIntestinal PAL-ka
110,18,058-
 20,8to 12.0240-
 32,016,045+
 48,016,0+
 54,016,041+
II10,86,082-
 20,88,0320+
 32,016,098+
 48,016,0166+
 58,016,0190+
III10,88,018-
 22,08,022-
 34,08,0102-
 48,0to 12.072-
 58,0to 12.0142-
IV10,88,020-
 20,88,028
 32,016,0115-
 44,016,035-
 54,016,0230-
V10,26,055-
 20,88,068-
 32,016,0105-
 48,016,0115-
 58,016,0118-

Compared with the waters of the river “Source” of these differences was 49-160% in the period. In the autumn of indicators of activity of urease in water that is contaminated runoff landfill had significant differences from the comparison on average by 15% when comparing them with the data about the activity of urease in water runoff clean bogs and 23% in comparison with indicators of the activity of the enzyme in the river “Istok”. At the end of the observation period urease activity is sharply reduced compared to summer and early autumn, b the La about the same level with the urease activity of river water, but superior to the latter in the waters flow from a “pure” (samples 4,5) wetlands at 40.4%, and the ratio of potential activity was lowest in these samples.

2. The synchronous nature of the dynamics of changes in the activity of urease in all the studied waters (samples 1-5)and the fluctuations of its level in the contaminated (samples 4,5) water (75 to 300 mg/l NH+3)- 2.3 times greater than the range of urease activity during the observation period in the unpolluted waters of the swamp, indicate a dominant influence of climatic conditions on the functioning of aquatic ecosystems, causing activity of the enzyme in water, which greatly enhanced by anthropogenic load.

3. Determination of the coefficient of potential enzyme urease activity showed the highest values (11 to 40) in river waters (both clean and polluted samples 1,2), mean values in the unpolluted waters of the swamp (from 6 to 20 sample 3), the lowest values of this relative index (from 4 to 10.9) in the waters flow landfill poultry.

4. Low values of the coefficient of the potential, the activity of urease in water runoff landfill during the period of observations at high values of indicators of enzyme activity (up to 300 mg/l NH+3St. Petersburg is there about the tension of the adaptation processes in the ecosystem of the swamp polygon. High values of the potential activity of urease in river waters (samples 1 and 2) 2.5-4 times exceed the values of the coefficients water runoff polygon characterize the sustainability of river ecosystems to pollution, which do not exceed the limits of its tolerance.

Parallel assessments were taken water samples by standard methods to determine the hygienic status of the species (table 2).

In table 2 data confirm the initial estimate obtained by the proposed method and show that in samples 4-5 oxidation (demand in mg/l of oxygen) large (16 mg/l) and higher than in control samples. This indicates pollution of wastewater by organic compounds, which can be a substrate for exposure and reproduction of microorganisms at their disposal, which shows the validity of the proposed method, the possibility of its use for assessing the state of water in the zone of flow selhozpredpriyatii, in particular poultry.

An important addition to presented is a comparison of the levels of NH3in the investigated waters by the method of colorimetry and commonly used in practice hygiene studies.

So, according to the data given in table 1 and 2, the content of ammonium compounds which deposits higher in samples 4 and 5 during the entire observation period. However, a higher sensitivity is used in the method colorimetric method allows to detect large values of the content of NH3in the waters, in 3-5 exceeding those of the same samples when determining the NH3the usual procedure.

1. The method of assessment of the water source area runoff agricultural enterprises, including water sampling, analysis, mathematical processing of the results of the analyses, characterized in that take water samples, at least in two areas - in the area of agricultural run-off businesses and area, which excluded the impact of runoff agricultural enterprises, in different time periods, conduct analyses to determine the content of ammonium compounds and enzyme urease-carbamidomethylated in water samples, and mathematical processing of the results is to compare the content of ammonium compounds and enzyme urease-carbamidomethylated in each water sample and comparison of the dynamics of change these indicators in water samples taken from different zones.

2. The method according to claim 1, characterized in that the sampling produce water at a depth of 0.3 to 1 m at a distance of 1-2 m from the edge of the shore, the water source is a river, and as areas where excluded the impact of the runoff of agricultural enterprises is ment, is the area at a distance of 1-2 km upstream from the confluence of runoff agricultural enterprises, and water samples taken at the same time.

3. The method according to claim 1, characterized in that the content of ammonium compounds in water samples to determine the photocolorimeter in parallel with the determination of the activity of the enzyme urease-carbamidomethylated.



 

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