Method for determining and forecasting the sanitary state of soil in the area of industrial pig

 

(57) Abstract:

The essence of the method is that when conducting soil analysis including content ammonitella microflora additionally conduct analyses to determine the activity of the enzymes catalase, nitrate reductase and urease. Collectively, the results of the analyses, there are three stages of stress of the soil, and remediation measures to restore soil appointed on the basis of the stage of stress of the soil. The technical result consists in the possibility of an integrated study of soil conditions and more effective forecasting of self-healing. 2 C.p. f-crystals, 3 ill., table 2.

The invention relates to agricultural production, in particular, to industrial pig production, mainly to soil Microbiology in the areas of industrial livestock production, and can be used in the environment to determine and predict the state of the soil buffer zones to mitigate the impact of farms on the environment and to issue recommendations on environmental activities in the adjacent areas, as well as for veterinary and zoo-hygienic control publ. 15.02.89. Bull. 6), which are determined by the level of soil fertility and the impact of soil contamination on the functional properties of the soil microbiota through the use of urea in an amount of 0.5 to 2.0% by weight of the soil as readily available for microorganisms, organic matter, and the assessment of the conduct on the rate of accumulation of ammonia and change the color of the indicator paper. At high speed this Express method disadvantages are its low accuracy (pH visually by the color of the indicator paper) and that is determined by only one factor that can not objectively assess the condition of the soil and its ability to self-clean in the conditions of anthropogenic impact.

Also known "Method of determination of biogenic features rock dumps of coal mines" (see A. with. N 920070, C 12 Q 1/06, publ. 15.04.82. Bull. N 14), wherein as a test condition of the soil in the dumps reclaimed breeds used seeding of microorganisms on the filter paper strips impregnated 3-5% meat-peptone agar containing the indicator 2,3,5-triphenyltetrazolium, replacement, according to the authors, their definition in Petri dishes for meat-peptone agar and reduce production time test the TES on solving a particular problem, that impairs its use, in particular when determining the environmental pollution in the vicinity of pig farms.

In addition, it is known that when the sanitary supervision of the soil conservation and health assessment of contaminated soils is generally complete the program, conduct the following laboratory tests:

1. Sanitary and physical (mechanical composition, humidity, filtration capacity, and others);

2. Sanitary-chemical (the ratio of protein nitrogen to total organic health number, pesticides, micro - and microelementary composition of the soil, heavy metals and others);

3. Sanitary-bacteriological (e-coli, the titer of Ana, titer of thermophilic bacteria and other);

4. Sanitary helminthological (helminth eggs).

5. Sanitary entomological (larvae and pupae of flies);

6. Radiometric.

In some cases laboratory program may be limited to individual indicators.

For sanitary-hygienic assessment of soil primarily examine the indicators that depend on pollution, typical for this area (see Gershon C. I., Boyko, I. A. Workshop on veterinary hygiene. Uch. manual./P> The disadvantage of this conventional program evaluation of soil pollution is that it (parts of sanitary-bacteriological studies) provides for the determination of the soil's ability to cleanse itself and restore normal functioning due to microbiological processes, without which it is impossible to predict the load on the soil from external contamination and to propose measures to clean up the soils, in particular in the further exploitation of pig farms, as only determined by the program indicator self-cleaning soil - titer of thermoflow conducted by sowing soil dilutions 1.5% meat-peptone agar does not give a comprehensive picture of the full activity of microflora and shows the ability of the microflora to withstand external pollutants.

In addition, according to the existing methods for carrying out sanitary-microbiological studies the selection of soil samples produced from a depth of 20 cm, which is true for cultural arable land, but somewhat distorts the overall picture of the activity of the microflora in the territory of pig farms, where the surface soil layer is not processed and the greatest activity of the microflora occurs only in the surface is nihayda and do not affect the purification of soil,

The objective of the invention is to develop a method of complex definitions and forecasting sanitary-microbiological and hygienic condition of the soil in the area of industrial pig, which allows using standard equipment veterinary laboratories versatile to determine biochemical, microbiological and sanitary condition of soil, their ability to self-healing and self-cleaning and to give sound recommendations for land reclamation security in sanitary-protective zone of the pig farm and the further effective use of them.

The task is solved in that in addition to the definitions in the soil samples taken in the given order in the surface soil layer, ammonifiers flora growing on meat-peptone agar, as well as levels of polluting the soil of nutrients - nitrate - and nitritional, ammonia, further define the parameters of biochemical processes in the soil, in particular the enzymatic activity of soil enzyme activities of catalase, nitrate reductase and urease. Then by comparing the activity of these enzymes with the level of content in soil decomposition products of polluting substances, nitrates and ammonia, pochwy, distinguishing three stages of stress soils that characterize the development in the soil "General adaptation syndrome" syndrome, specific nature of the responses, manifested in changes in the biological activity of the surface layer of soil in the sanitary-protective zone of pig farms, and non-specific in origin (different according to the chemical nature of the pollutants). It is believed that in the first stage of stress soil visible changes occur that indicate a serious breach of natural biological processes in the surface layer of the soil. In the second stage is the protective mobilization reserves of soil microflora (biota) and the restoration of depleted stocks of protective forces of a living organism, which is the soil.

If the action of damaging factors continues, then comes the third stage of stress, which is contained in the soil microorganisms and the soil loses its resistance, lose adaptation, resulting in come the depletion and destruction of the soil loses its ability to self-cleaning and restoration.

Depending on the specific stage of stress soils are assigned to measures for remediation of soils or to increase its JV is in the sanitary-protective zone of the pig.

The method is as follows.

Upon arrival to the object of investigation (pig) delineates the point of taking soil samples. The first point is determined directly about the premises, which contains svinopogolovje, but not on the road or a paved area, and in the place where there may be some herbaceous vegetation. The following point sampling occur within 50 meters in the direction of the prevailing winds, or on the water. A distance of 50 meters adopted by the results of practical experiments as providing requiring the accuracy of the forecast and do not require a very large number of measurements. The last samples are taken at a distance of 350-500 meters depending on the size of the designated buffer zone and customer requirements.

Simultaneously with the taking of soil samples in the target sanitary zone take control soil samples of the same type, but outside the buffer zone at a distance from the inspected object in 3-5 miles away, in places where there is no effect of the pig, but the same soil and natural conditions. The mass of the soil sample is taken of the order of 0.5 kg according to standard techniques adopted in soil science, in particular, as described in the book: the Workshop is delivered to the laboratory for analysis and research.

Processing samples in the laboratory is carried out by standard methods, depending on specified programs. In addition to the conventional parameters further define the enzymatic activity of soils, in particular using methods for the determination of enzymes in soil modification A. W. Galstyan (1974). These methods are defined catalase (H2O2: H2O - oxidoreductase, EC 1.111.6), urease (urea - amidohydrolase, EC 3.5.1.5), nitroreductase (restored ON: nitrate oxidoreductase EC 1.6.6.1).

The results of all measurements and investigations handled by methods of mathematical statistics are in tables and presented in graph form in the course of sampling, indicating the control options. Then the analysis of sanitation in the area of the pig with the definition of the zones of stress state of the soil, highlighting three areas of stress, depending on the totality of the results obtained. If necessary, assign additional research and identifies measures for remediation of the soils in the sanitary-protective zone of the pig.

Example determine and predict the sanitary sachweh insemination of plempredpriyaty "Sverdlovsk", located in the village. "Red October" Bogdanowitsch district, 80 km from Ekaterinburg. Production direction of the economy - grunts-producers and breeding pigs. The plot, which took soil samples, smooth with a slight slope towards the pond (400 m), well-lit by the sun, covered with grassy vegetation, soil, sod-podzolic, the most typical of the region.

Soil samples were taken at depths of 0-5 cm, in five replicates on each of the plots, located at a distance of 50 m from each other in the direction of the prevailing winds (North-East), taking into account the fact that pollutants accumulate predominantly in the upper horizon of the soil. As a control used soil samples from sites located at a distance of 5 km from the farm, not polluted by industrial wastes with similar physical-chemical properties and the nature of the vegetation on them with the studied soils.

Processing samples of the soil were carried out in the laboratory of Department of Microbiology, sanitation and hygiene Ural state agricultural Academy, according to standard techniques and, in chastest the chemical soil analysis", S. 4 and the Enzymatic activity of soils", S. 72-95).

The results of studies after mathematical processing are summarized in table N 1 and N 2, and presented in graph form in Fig. 1, 2, 3.

In table No. 2 shows the stage of "stress" of soil are defined according to the proposed method and the known theory, Selye.

According to this theory in the first stage visible changes occur in the body (in the soil), indicating a serious breach of natural biological processes under the action of the stressor. To restore the normal functions of the body in the second stage be mobilization of all its protective reserves and the restoration of depleted stocks of protective forces of an organism. If the action of damaging factors continues, then comes the third stage, in which a living organism (soil) loses its resistance, loses adaptation, the result is exhaustion and even death.

In other words, when stage I adaptation syndrome soils are in the initial stages of stress, and greater adaptive capacity for recovery and stabilization.

When stage II adaptation syndrome soil are the mi skills to restore damaged by pollution of the ecosystem.

Stage III indicates profound violations and irreversibility of soil-forming processes.

In Fig. 1-3 shows the dynamics of changes in the parameters studied in the form of graphs depending on the distance from the object contamination.

On graphs with index - and - shows the parameters common parameters: nitrite, acidity (pH) and oxidation.

On graphs with index - b shows the parameters: nitroreductase, urease, ammonia, catalase and flora.

After a comprehensive analysis of the obtained results (table 1 and 2) and using graphs (Fig.1-3), we can draw the following conclusions. The maximum saturation of the microorganisms is characterized by a soil test of the area remote from the source of contamination at 250 m (table. 2). The number of microorganisms of ammonification reaches 6, 2, 10 in 1 g of dry matter. The minimum content of the microorganisms identified in the SPT 50 m - 1.0.10 on 1 g of dry matter. Several other trends are observed in the enzymatic activity of soil survey sites (table. 2). The soil on the SPT 150 m of nitrate reductase (as well as a high content of microflora - 5.5.10 1 g of dry matter), whereas the lowest catalase activity identified on SPT 200 m, the s as the distance from the pollution source waste pig revealed a number of natural vzaimoobuslovlennykh processes (Fig. 1-3), indicating different degrees of tension adaptive responses of soil biota in response to the harmful effects of pollutants. Thus, a high level of activity of enzymes nitroreductase and urease on the SPT 150 m is accompanied by the most high content of nitrate - and nitritional, ammonia, a sharp decrease in the content of hydrogen sulfide, which indicates that the most pronounced decomposition of organic matter entering the soil, inducing increased redox reactions in it, as evidenced by the high (six times superior to the activity of the control soil samples) catalase activity. Since the content of hydrogen sulfide in closer to the pollution source (50-100 m) and remote areas (SPT 300 and SPT 350 m) is significantly greater than that on the SPT 150 and the 200 m and the oxidation of the soil, indicating the presence of organic substances in it, also the highest on the site 250-350 m from the source of pollution, therefore, the processes of decomposition of organic matter in these soils are less intensively than in the area of 150-200 m the Results of this study suggest the processes occurring in soils affected by pollution from (stress), and in these soils can be selected stages of stress.

Thus, as a result of the research showed that the soil in the sanitary-protective zone of pig enterprises subjected to prolonged exposure to pollution from pig production, are under stress, or "General adaptation syndrome" syndrome, specific nature of the responses, manifested in changes in the biological activity of the soil, and non-specific in origin (the action of various chemical nature of contaminants).

Application in the research of indicators of biological activity in soil (content of microflora and enzymatic activity) and compare them with levels of pollutants allowed to detect the stages of the adaptation syndrome in soils pig farms. Thus, a plot that is more intensive decomposition of organic substances, according to theory of stress, is in the condition stage II adaptation syndrome (150-250 m). The state of the I stage of stress identified in the research in soils, remote from the pollution source at a distance of 300-350 m, because in this area low sod is located to the source of contamination section 0-100 m, characterized by low activity of enzymes and content of microflora and a high content of contaminants (hydrogen sulfide, ammonia, nitrate, and nutritional) noted the development of the third stage of stress. Soils in areas of anthropogenic impact in the development of stage III of stress are particularly in need of remediation activities, which are determined in accordance with the needs of their future use.

It is established that under the influence of polluting substances of different chemical nature, the sources of which are, in particular, pig enterprises occur in varying degrees expressed changes and disturbances of soil biological activity (enzymatic and microbiological) depending on the distance and direction from the source of contamination. The activity of oxidative enzymes in soils under pig enterprises varies depending on the number of accumulated pollutants. With the increase in the build up to a critical level in soil, as a rule, the reduced activity of the enzymes catalase, urease and nitroreductase.

To assess the sanitary condition of the soil and its ability to sa is ugrasena waste pig, it is suggested to conduct a comprehensive study that includes in addition to the commonly used methods of determining the activity of enzymes: catalase as a test on the intensity of oxidative processes in the soil urease and nitroreductase as indicators of activity of the process of self-purification of soil from contaminants.

Based on the analysis of the results of a comprehensive study on the changes of oxidative processes in soils in different zones of contamination, as measured by the level of activity of catalase, urease, nitroreductase, and the number of microorganisms of ammonification (growing MPa) formulated a hypothesis about the development of the adaptation syndrome (stress) in soils under the influence of polluting waste hog substances of different chemical nature. The state of microbiological activity and oxidoreductase activity (catalase, urease and nitroreductase) in the soil has identified three stages of stress, each of which is defined two phases of its development, with a gradual transition from one to the other.

In particular, in the area of medium contamination (on the destruction of 350 m and 300 m from the pigsty), the first and second phase stage I stress, characterized by the absence of amplification okisliteljno and enzymatic activity. In the zone of strong pollution from pig production (at a distance of 250 m) revealed the first phase II stage of stress, characterized by a sharp increase in the number of microorganisms of ammonification and changes in indicators of oxidative processes in the soil compared with the control. On the destruction of 100-200 m from the pollution source in the presence of high-level accumulation of polluting the soil of substances selected the second phase of stage II of the stress at which a sharp increase of oxidative processes in the soil. Inside the pigpen, the first, and at a distance of 50 m from it - the second phase III stage of stress with a significant reduction in the number of microflora and intensity of oxidative processes in the soil (the second phase).

The proposed method using the method of bioindication and determining the enzyme activity to assess the sanitary condition of the soil on the farm confirmed the appropriateness of the method because it is useful for the diagnosis of the physiological condition of the soil, and to evaluate the performance of treatment facilities and the effectiveness of measures for decontamination of the soil in the areas of pollution from pig production. Analysis of R is the SQL development process of adaptation in them and promptly recommend carrying out reclamation activities.

1. Method for determining and forecasting the sanitary state of soil in the area of industrial pig, including the taking of soil samples, prepare samples, conduct tests, including the content ammonitella flora growing on meat-peptone agar, mathematical processing of results and conclusion about the state of the soil in the area of the inspected object, characterized in that it further conduct analyses to determine the enzyme activity in soil: catalase (H2O2:H2O - oxidoreductase. KF I. III.6), nitrate reductase (restored ON: nitrate oxidoreductase. KF I. 6.6.1) and urease (urea-amidohydrolase. EC 3.5.1.5), and specify the contents in soil decomposition products of polluting substances, nitrates and ammonia, and collectively, the results of the analysis highlight in stage stress of the soil, characterized by the activity of enzymes produced in the process of vital activity of the soil microflora, and reclamation measures to restore soil are assigned based on the stage of stress of the soil.

2. The method according to p. 1, characterized in that the sampling of the soils in the survey area, which carried the Oia, moreover, as a control option when studies take soil samples of similar composition from the same area, but no closer than 5 km from the inspected object, taken simultaneously with the main samples.

3. The method according to p. 1, characterized in that the content of ammonia and nitrate ions in soil samples to determine the methods used to determine the activity of the enzymes urease and nitrate reductase, and instrument method on photocolorimetric in parallel with the determination of the enzyme activity.

 

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