SU920070A1 - Method of determining biogenic properties of coal mine and dump muck - Google Patents

Description

(54) METHOD FOR DETERMINING THE BIOGENEITY OF BREED DEPOSITS OF COAL MINES

The invention relates to rural and forestry, in particular to the determination of the biogenicity of industrial waste dumps during their biological reclamation. A known method for indicating the microorganisms of the group of Escherichia coli in water is on nutrient pads made of sheet filter paper impregnated with Hotinger's m-peptone broth with an indicator of tetrazolium chloride. The cultivation of microorganisms is carried out at 3037 for 10-1 hours. The disadvantage of this method is that it allows only bacteria of the group of Escherichia coli to be detected and is used as a guide, since it has a large definition. There is a known method for indicating bacteria on agar media in Petri dishes (Koch method), which consists in preparing a series of dilutions of mac-peptone agar in sterile test tubes; limiting dilutions), after which a certain volume (1 ml) of the test suspension is sown in Petri dishes filled with (10 kp) m-peptone agar. Bacteria are grown in a thermostat at 30 ° C for 2–3 days, then the number of grown colonies 2 is counted. The disadvantages of this method are the high consumption of the nutrient medium, a large number of special dishes and the duration of the determination, which creates certain inconveniences in the expeditionary conditions. The purpose of the invention is to accelerate the determination of the biogenicity of the rock dumps of coal mines during biological recultivation. The goal is achieved by planting a suspension of rocks produced on strips (filter paper

soaked with m with peptone agar containing indicator 2,3,5-triphenyltetrazolium chloride.

Saprophytic microorganisms that develop on m-peptone agar give s an idea of the total biogenicity of the soil, since microorganisms growing on m-peptone agar are part of various communities (microorganisms that decompose the organo-oral complex rocks) and are a diagnostic indicator of the biogenicity of dump rocks being reclaimed „

The biogenicity indication is used for de-ashed ash filters with white entoy (medium filtering), during the impregnation of which the m-peptone agar is distributed to the filter through the filter. When using deashed filters with th blue tape (slow filtering). and red tape filters (fast filtering), as studies show, m-peptone agar is not evenly distributed, which affects microbial growth and affects the accuracy of analyzes.

On the basis of experimental data, it was established that on indicator nutritional papers, impregnated with m-peptone agar in a concentration of less than 34, the growth of saprophytic microorganisms is not detected, since there is not enough nutritional substrate to develop them. M so-peptone j, ny agar in a concentration of more than 5 is not absorbed by the filter paper.

Growing saprophytic microorganisms on indicator nutrient loosening provides a quick determination of the biogenicity of the breed within 5-9 hours.

The test procedure is as follows.

Filter paper strips (2x8.5 cm) (deashed filters with white tape) are kept in a box with germicidal lamps for 10 minutes (5 minutes on each side). M co-peptone. agar sterilized

in the vail for 20 mins at 120 C. The aqueous solution of the indicator 2,3 5-tri-, eniltetrazolium chloride in the concentration is sterilized in test tubes separately. Then it is poured to the g / co-peptic agar until the concentration of the indicator is 0.2. Filter strips attached by a holder to

tripod, immersed in sterile and peptone agar with indix-ztor. At the same time, filter paper strips absorb mc0 peggy agar for 5 - -wO, 6-0 ,,. Soaking of filter paper strips is made when irradiators are on. The strips of propium filter paper are placed on sterile paper and dried at B ° C for c in a drying cabinet. A suspension of rock (1 g) in sterile water (100 ml) is prepared and the strips of the obtained indicator paper S are impregnated with it for c, sealed in sterile plastic bags and placed in a thermostat at 28-30 ° C. After 5 hours, the growth of bacterial colonies is counted with a microscope.

Example 1. The toxicity of a toxic rock is determined from a dump of a charge consisting of argillites, aleurolites and quartz sandstones with pyrite inclusions and containing the following elements, /;: total carbon 9, total iron 7.7; total aluminum 8.7; total nitrogen 0.21; total sulfur 1.1 phosphates, 92; potassium O, (J i; copper 0.002; boron 0.17; manganese 0.27. Samples of the rock are taken before restoration works and after the breeding of the breed from the depth, in cm: 0-5, 5-10, 10-20. Then it is ready t water slurries, at the rate of 1 g of rock per 100 ml of sterile distilled water.

Cultivation and indication of saprophytic microorganisms is carried out on indicator papers, impregnated with 5% m-peptone agar with indicator 2,3) triphenyltetrazolium chloride, as described above. The growing time of microorganisms on the indicator paper 5 hours

Example 2: The biogenicity of rocks is determined using indicator paper impregnated with 2; -Mo-pept (Onn agar, with indium (2.3) 5-triphenyltetrazolium chloride. Microorganism growth time is 7 h.

Example 3. .. Determine the biogenicity of the rocks by growing microorganisms on indicator papers impregnated with 3 ° green and co-peptone agar with an indicator of 2,3,5-trifeiyl tetrazolium chloride. Microbial growth time 9 h.

Table 1 shows the data of examples 1–3) on the determination of the biogenicity in the initial dump rock before recultivation, proposed by the method and method with the microorganism growth in Petri dishes.

From the data in Table 1, it can be seen that the original dump rock is characterized by .jj.

Table 1

low yiogennost, i.e. the rock contains a small amount of saprophytic liquor organisms, which is confirmed by the low content of humus, nitrogen and phosphorus in the rock. In tab. 2 presents the data (in examples 1-3) to determine the biogenicity of the dumped breed, subjected to biological reclamation.

table 2

From tabla 2 it can be seen that in the dump rock subjected to biological recultivation, the biogenicity increases significantly, the number of microorganisms increases, the content of humus, nitrogen and mobile phosphorus increases.

Coincidence of the results when counting the number of saprophytic microorganisms grown on indicator papers and in 95-97 ° Petri dishes. Optimum results were obtained when growing microorganisms on indicator papers impregnated with m-peptone A1 agar, incubated in a thermostat for 5 hours.

Thus, the determination of microorganisms in rocks with the help of indicator papers can replace their definition in Petri dishes on m and septon agar, which eliminates the need to prepare a large amount of nutrient medium and sterile dishes. In addition, the growth time of microorganisms is reduced and the need for such time-consuming definitions as the determination of humus, nitrogen, phosphorus is eliminated.

The proposed method of determining the biogenicity of rocks can be used in expeditionary conditions, it is necessary to decide the question of the suitability of otvapov coal mines for reclamation and to assess the intensity of the soil-forming process on the surface of the dump during biological

reclamation.

invention formula

The method for determining the biogenicity of a rock dumps of coal mines by preparing a suspension, sowing on a nutrient medium, incubating and counting colonies of microorganisms, is characterized in that

the purpose of speeding up the determination process, seeding is carried out on strips of filter paper impregnated with m-peptone agar containing

indicator 2,3 5-triphenyltetrazolium chloride

Sources of information taken into account in the examination of 1- Korsh L, E, Lrtemova G.Z Accelerated methods of sanitary bacteriological examination of water, Moscow, Medicine, 1978, p. 158-165,

2, Workshop on Microbiology, Ed. By N., Ssegorova, Moscow State University Publishing House, 1976, p. 61-63,

Claims (1)

Claim The method of determining the biogenicity of the breed of dumps of coal mines by preparing a suspension, seeding on a nutrient medium, incubation and counting colonies of microorganisms, characterized in that, in order to accelerate the determination process, sowing is carried out on strips of filter paper impregnated with 3 “5% ~ meat-peptone agar containing 20 indicator 2,3,5 ^_ triphenyltetrazolium chloride.