Biopreparation for treatment of water, industrial drains and soil from pesticides resistant to decomposition and method of its application

FIELD: biotechnologies.

SUBSTANCE: invention may be used to produce a new biopreparation for cleaning of water, soil, industrial drains from pesticides resistant to decomposition and selected from chlorophenoxyacetic acids, such as 2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T), chlorophenoxyacetic acid (CPAA), phenoxyacetic acid (PAA), - 2,4-dichlorophenoxy-α-propionic acid, 2-methyl-4-chlorophenoxy-α-propionic acid, 2,4,5-trichlorophenoxy-α-propionic acid, 2,4-dichlorophenoxy-α-butyric acid, methyl-[1-(butylamino) carbonyl]-1H-benzimidazol-2-ylcarbamate, 2,4-dichlorophenol, imidoclapride, hexachlorohexane, and also phenol. The biopreparation is an association of strains of bacteria Pseudomonas putida VKPM V-10997, Bacillus cubtilis VKPM V-10999 and Rhodococcus erythropolis VKPM - As-1882 at the weight ratio of (1-2):(1-2):1. At the same the biopreparation, as a rule, contains additionally a sorbent, organic, mineral and stimulating additives and has activity that stimulates growth of plants and fungicide properties. The produced biopreparation is introduced as an aqueous solution by means of sprinkling into polluted soil or industrial drain in efficient amount.

EFFECT: invention makes it possible to improve cleaning of water, soil, industrial drains from pesticides resistant to decomposition.

21 tbl, 29 ex

 

The invention relates to a new biological product for the treatment of water, soil and industrial waste, contaminated resistant to degradation by chemicals classified as hazardous, such as widely used in agriculture pesticides.

Biologics-biodestruction and their use for cleaning soils from oil and oil products containing Bacillus brevis and Arthrobacter species described in the patents of the Russian Federation 2323970 and 2237711, and in the patent of the Russian Federation 2086667 described the consortium, including microorganisms Pseudomonas putida and Bacillus subtilis.

Also known biological products "Roder" on the basis of strains of Rhodococcus ruber VKM AC-D and Rhodococcus erythropolis VKM AC-D, "Lenoil", "Devoroil", "Acabal" on the basis of bacterial strains strains of Pseudomonas and yeast, which are effectively used for removal of aliphatic fractions of crude oil.

Known decomposition of chlorinated aromatic pesticides (2,4-dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid) using Pseudomonas pseudoalcaligenes strain NRRL B-18087 (U.S. patent 4804629), detoxification of residues of organophosphorus pesticides and their toxic metabolites by strains of the bacterium Agrobacterium radiobacter (ed. mon. The USSR 1250572). In AVT. mon. The USSR 1560487 describes how the degradation of the pesticide 3,4-dichloraniline algae Chlorella vulgaris BM-a-10 and Scenedesmus acuminalus UA-2-7a. Decomposition of 2-chlorbenzoyl acid bacteria Pseudomonas putida YNK-1 is described is seen in U.S. patent 4803166; phosphate impurities and chlorinated hydrocarbons by microorganisms Moraxella and Arthrobacter separately or in combination - in the Federal Republic of Germany patent 3729127. Known degradation of compounds such as DDT, polihlorvinila, benzopyrenes, dioxins, using basidiomycete Phanerochate chrysosporium (U.S. patent 4891320). The strain Pseudomonas putida - 106 is an active destructor of dimethylphenylcarbinol and phenol (3), the bacteria Pseudomonas pseudoacaligenes destroy aromatic and heterocyclic compounds, most often found in wastewater (5), and a strain of Pseudomonas pseudoacaligenes destroys the aromatic compounds in solid and liquid medium (4). Ability to degradation of chloropropionic triazine, such as Simazine and atrazine, have some Museum strains of Nocardia, Arthrobacter, Micromonospora (6). In U.S. patent 6632363 describes a composition comprising a hydrophobic medium and the bacteria Bacillus subtilis, and the method of its use to improve water quality, containing, for example, pesticides.

Ksenofontova O.Y, etc. in the article "changes in the populations of soil microorganisms under the influence of pesticides" (Izvestia, Saratov state University, Scientific. Section "Chemistry, biological. Ecology, 2007, Vol.7, No. 2, p.66) describes as the most active destructors strains of aerobic bacteria of the genera Bacillus and Pseudomonas, destroying the following pesticides: chlorthiazide, uglon, semifinale, nitrolon, carotid and CT is the te.

In the patent of the Russian Federation 2410170 describes a method of cleaning contaminated soil from organic compounds, including pesticides, for example from dichlorobiphenyl (DHB), by introducing sorbent, representing glauconite breed, pre-treated at 200 to 300°C., and a strain of bacteria of the genus Rhodococcus.

All of these methods of detoxification of pesticides (toxic chemicals) for the decomposition of specific chemical compounds offered certain types of microorganisms. Describes mainly the decomposition of pesticides in the aquatic environment when using xenobiotics by microorganisms as the sole source of carbon and energy, or require the additional use of sorbents.

A significant drawback of all these cultures is their narrow specificity to a specific substrate, and they do not have the preparative form and are not applied in practice for cleaning contaminated recalcitrant compounds such as pesticides, water, industrial wastewater and soil.

In the patent of the Russian Federation 2093478 described the Association of bacterial strains Pseudomonas putida BKM 1301, Bacillus subtilis BKM B-1742D and Rhodococcus erythropolis BKM Ac-1339D in the ratio of 1:1:1 for the purification of water and soil from oil and oil products and polymer additives in the drilling fluid and its use. However, the use of the above Association to clean the key water soils and industrial effluents contaminated resistant to degradation by chemicals, such as pesticides, previously was not known.

Closest to the preparation of the present invention the target is a biological product "Biava" (patent of the Russian Federation 2248255), which contributes to the improvement of soil fertility and to stimulate the natural microflora of the environment. The inoculant contains amylolytic, proteolytic and nitrogen-fixing organisms, such as microorganisms of the genus Pseudomonas, Bacillus. The disadvantage of this drug is a complex composition that includes more than 25 different kinds of organisms that are not adapted to associative interactions, which is difficult to manufacture and to maintain strains. In the mentioned patent also describes the use of a biological product "Biava" the introduction of the latter into the soil.

This drug is not a stimulant plant growth and does not have the degradability of these persistent chemicals such as pesticides or herbicides from a number chlorophenoxyacetic acid: 2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5,-T), phenol, 2,4-dichlorphenol, HPUK and other chemical compounds, as described below. These compounds are among the most stable and persistent in the environment. The ingestion of these chemicals is of such substances can lead to serious diseases, tissues and nervous system. The presence of their food is considered invalid (see, for example, Tinsley I. the Behavior of chemical contaminants in the environment. TRANS. From English. M.: Mir, 1992, s).

The present invention is to develop a new biological product for cleansing (detoxification) soil, water and industrial effluents contaminated with persistent chemicals, such as pesticides, phenols, and how use of the drug. Drug FENOX according to the present invention improves the cleaning of the environment from toxic compounds and, consequently, increases the consumer quality products grown on treated land. When this happens microbial degradation of toxic substances by the cells of the bacteria in the soil to non-hazardous compounds. Biological product shows high efficiency in wastewater treatment of industrial enterprises.

The present invention relates to a biological FENOX for cleaning of ground, soil, industrial waste water from decay-resistant chemicals, such as pesticides, which is an Association of new strains of bacteria Pseudomonas putida. Bacillus subtilis and Rhodococcus erythropolis mass ratio of (1-2):(1-2):(1), which can optionally contain a sorbent or immobilized sorbent, mineral, organic and stimulating supplements.

Difference p is izlagaemogo drug "FENOX" from previously known nearest analogue to destination is specified Association strains of bacteria in a specified mass ratio.

The drug is particularly effective when it is used for decomposition of persistent pesticides, selected from the group chlorophenoxyacetic acids, such as 2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T), 2,4-dichlorophenoxy-α-propionic (Dichlorprop, 2,4-AL), 2-methyl-4-chlorophenoxy-α-propionic (Mecoprop, 2M-HP, MSRR), 2,4,5-trichlorophenoxy-α-propionic (2,4,5-TP Silvex), 2,4-dichlorophenoxy-α-oil (2,4-DV); methyl-[1-[(butylamino)carbonyl]-1H-benzimidazole-2-yl]carbamate and the drug based on it (benomyl, carbendazin, benzol); Isidor, where the active ingredient is a neonicotinoid Imidacloprid, imidacloprid; contran, where the active substance is metribuzin (metribuzin), HCH (hexachlorocyclohexane, HCH, exatons, damix, sinex), HFUK (chlorprothixene acid), FOOK (venexiana acid), hexachlorophene, 1,1-di(4'-chlorophenyl)-2,2-dichloroethane (DDT, dust), and 2,4-dichlorophenol and phenol. Especially the drug is effective when it is used for decomposition of persistent pesticides from the group chlorophenoxyacetic acids, such as 2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T), chlorophenoxyacetic acid (HPUK), venexiana acid (FOOK), and 2,4-dichlorophenol and phenol, Imidacloprid.

The drug of the present invention in contrast to previously known drugs the the ATA has advanced growth-stimulating effect on the germination and growth of cultivated plants, it also has fungicidal activity.

The object of the invention is also the method of application of the specified biological product "Fenox". The method consists in introducing an effective amount of the drug in contaminated soil, soil or industrial runoff.

The difference of the proposed method of application from the previously known closest analogue is the introduction of the Association of these bacteria strains in the specified mass ratio.

Preferably the drug is administered in an amount of 10 kg of dry matter at the time of ploughing based on 1 ha of land at ambient temperatures, mainly at a temperature of 15-35°C. The most preferred methods of administration in the form of an aqueous solution by sprinkling. You can also make the introduction of the drug into the soil by the introduction of seeds treated with the suspension of the drug with the addition of nutrients, such as vermicompost, or the introduction of a dry formulation of the drug in powder form into the ground at the time of ploughing or sowing seeds.

Any drug can be sown in the soil seed treated with the suspension of the drug with the addition of nutrients, such as vermicompost.

Pre-preparation plants in the tank of water with the addition of vermicompost and nitrogen-phosphate fertilizers.

It is also possible introduction of dry preparation the second form of the drug in powder form into the ground at the time of ploughing or sowing seeds. Proposed according to the present invention the drug FENOX" have a stimulating effect on the growth of plants and strains of the drug good prizhivaniyu in the soil, which leads to efficient detoxification of the environment. Strains of Pseudomonas putida and Rhodococcus erythropolis are D-plasmid degradation and capable of transferring property destruction, pesticides, and other harmful substances indigenous bacterial populations in soil, water, wastewater, thereby increasing the cleaning of pollutants. The advantage of using this drug is the decomposition of pesticides in soil for up to 86% and in liquid medium up to 99%.

The current basis of preparation - the Association of soil bacterial strains Pseudomonas putida, Bacillus subtilis, Rhodococcus erythropolis. All cultures have polysubstance specificity to multiple xenobiotics: chloroperoxidase acids, such as 2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T)phenol, 2,4-dichlorphenol, HPUK and some others mentioned in the examples. The mixture gives a strengthening of the functions of cleaning the environment from toxic compounds. As sorbents used kaolin and melkoizmelchennye peat, which simultaneously serves as an organic additive, as organo-mineral and growth-stimulating supplements vermicompost extract from it and chitin-containing substrates. Before applying to the expansion of the t (bucket, the flask) dry powder product is well mixed in warm water (25-30°C) until a homogeneous suspension. 10 g of the drug stir in 10 l of water. It is recommended to add NPK (5 g/10 l) or compost (5 g/10 l), periodically the suspension is stirred for aeration and activation of microorganisms within 1-3 hours before use (watering from a watering soil (10-20 l/10 m2or processing of 100-150 ml/100 g seeds).

Obtaining a biological product is implemented by a separate submerged culture usually within about 20-30 hours 3 strains of these highly effective bacteria on standard biotechnological equipment. Seed produced by growing the strains on synthetic medium with the addition of the pesticide and use this material for planting fermentors large amount.

Accumulated dry biomass of cells is mixed in the ratio (1-2):(1-2):1. The overall title is not less than 5×108CFU/g dry product after adding media-sorbent and other additives (original concentrate 1010-11and not less than 5×108CFU/ml of the liquid preparation. For the industrial manufacturing and packaging, we offer drug uses generic biotech and filling equipment and materials.

Biological product eliminates the penetration into plants, vegetables and berries chemical means of plant protection, STI is ed on the best germination and growth of plants, inhibits fungal phytopathogens.

Finacom handle contaminated with chemicals, such as pesticides, phenol, possibly oil, water, industrial effluents, soil, and plant seeds.

The strains belonging to Fenox are new.

The Bacillus subtilis strain selected by the method of cumulative cultures from samples of soil industrial zone enterprises khimicheskoi industry of the southern Urals. The Bacillus subtilis strain deposited in Russian national Collection of Industrial Microorganisms under the number VKPM B-10999.

The strain of Bacillus subtilis identified based on the analysis of the nucleotide sequence of the 16S rRNA gene. The nucleotide sequence (length not less than 500 base pairs) DNA fragment encoding the gene for 16S rRNA, has similarities with the stated form, constituting 99%.

Cultural morphological traits

The strain of Bacillus subtilis is typical for this kind of Cultural-morphological characteristics. Sticks, the size of 0.2-0.5 1.5-3 μm, solitary, often in pairs. Form an oval and rounded endospores. Flagella are pietragallo. With the growth in meat-peptone broth culture forms developed matte film, in this case the medium is transparent. With shaking biomass in a liquid medium to its full dispersion does not occur. Colonies of dry fine-wrinkle, edge wavy. On the slices of potato growth is bilen in the form of a cream-colored colonies smooth, strongly folded. The color of Gram-positive.

Physiological and biochemical characteristics

The Bacillus subtilis strain grows well on glucose-peptone medium, LB agar and broth, glucose-mineral medium at a temperature of 28°C. Grow on media with L-Proline, DL-leucine, α-Ketoglutarate, DL-α-alanine, L-glutamine, D(+)-xylose, L-asparagine, chitin, DL-serine, glucose, phenol and chlorphenoxamine acids as the sole carbon source. The strain of Bacillus subtilis can be stored in a dried condition. Check the viability of the strain is carried out by plating on glucose-peptone agar, M9 and LB - 1 every 12 months. When stored on the shoals of these environments at 5°C change 1 times per 3 months.

The Bacillus subtilis strain is non-pathogenic and non-toxic microorganism. Intravenous infection of white mice did not show zoopathogenic properties.

The strain Rhodococcus erythropolis selected by the method of cumulative cultures from samples of soil industrial zone enterprises khimicheskoi industry of the southern Urals. The strain Rhodococcus erythropolis deposited in Russian national Collection of Industrial Microorganisms under the number VKPM As-1882.

The strain Rhodococcus erythropolis identified based on the analysis of the nucleotide sequence of the 16S rRNA gene. The nucleotide sequence (length not less than 500 base pairs) DNA fragment, the code is its 16S rRNA gene, has similarities with the stated form, constituting 99%.

Cultural morphological traits

The strain Rhodococcus erythropolis is typical for this kind of cultural-morphological characteristics. Cells are rod-shaped, rarely branching, diameter is 0.6-0.8 mm, length - 3-8 μm. Cells moderately polymorphic, are often in the form of V-forms are pronounced cycle of development chock-wand-cock: at the age of 24-36 hours of the rod-shaped cells begin to grow and increases the proportion of coccoid and oval cells. Mature colonies are pigmented, have orange paint. The consistency of a pasty colonies. The color of Gram-positive.

Physiological and biochemical characteristics

The strain Rhodococcus erythropolis grows well on glucose-peptone medium, LB agar and broth, glucose-mineral medium at a temperature of 28°C. Grow on media with L-Proline, D-arabinose, DL-α-alanine, L-glutamine, D(+)-xylose, L-asparagine, DL-serine, casein, glucose, phenol and chlorphenoxamine acids as the sole carbon source. The strain Rhodococcus erythropolis can be stored in a dried condition. Check the viability of the strain is carried out by plating on glucose-peptone agar, M9 and LB - 1 every 12 months. When stored on the shoals of these environments at 5°C change 1 times per 3 months.

The strain Rhodococcus erythropolis is non-pathogenic and n is a toxic microorganism. Intravenous infection of white mice did not show zoopathogenic properties.

The strain Pseudomonas putida selected by the method of cumulative cultures from samples of soil industrial zone enterprises of chemical industry of the southern Urals. The strain Pseudomonas putida deposited in Russian national Collection of Industrial Microorganisms under the number VKPM B-10997.

The strain Pseudomonas putida identified based on the analysis of the nucleotide sequence of the 16S rRNA gene. The nucleotide sequence (length not less than 500 base pairs) DNA fragment encoding the gene for 16S rRNA, has similarities with the stated views of 99%.

Cultural morphological traits

The strain Pseudomonas putida is typical for this kind of cultural-morphological characteristics. Moving sticks with polar flagella located. The size of the cells is 1-4 1.3 to 5.4 μm. Colonies on meat-peptone agar grayish color, within colonies and their reverse side there is a reddish-brown pigment. The dispute is not formed. The consistency of the colonies - viscous. The color of Gram-negative.

Physiological and biochemical characteristics

The strain Pseudomonas putida grows well on glucose-peptone medium, LB agar and broth, glucose-mineral medium at a temperature of 28°C. Grow on media with DL-tyrosine, DL-tryptophan, D(+)-xylose, D(-)-mannitol, L-glutamic acid, DL-Le is the Qing, α-Ketoglutarate, DL-α-alanine, D-ribose, L(+)-arabinose, L-glutamine, cellulose, L-asparagine, chitin, DL-serine, pectin, glucose, casein, phenol and chlorphenoxamine acids as the sole carbon source. The strain Pseudomonas putida may be stored in a dried condition. Check the viability of the strain is carried out by plating on glucose-peptone agar, M9 and LB - 1 every 12 months. When stored on the shoals of these environments at 5°C change 1 times per 3 months.

The strain Pseudomonas putida is non-pathogenic and non-toxic microorganism. When intravenous infection of rabbits does not show zoopathogenic properties.

Example 1.

Detection of plasmid degradation of pesticides in strains

The destructive properties of bacteria can be determined extrachromosomal elements [Don, Pemberton, 1985; Ghosaletal., 1985]. Such elements - plasmids detected in strains of Pseudomonas putida In PMBC-10997 and Rhodococcus erhytropolis PMBC Ac-1882.

By the method of alkaline lysis (Maniatis T., Fritsch E., Sambrook j. Methods of genetic engineering. Molecular cloning: TRANS. from English. Ed. Bev. - M.: Mir, 1984. - 480 C.) distinguish them from the cells of these strains and then the drugs plasmids fractionary agarose gel under standard conditions (Maniatis et al., 1984] (see figure 1 - Plasmid profile of strains: 2-9 Pseudomonas putida VKPM B-10997, 10-17 Rhodococcus erhytropolis PMBC Ac-1882).

Note the R 2.

Compatibility of strains included in the preparation

The absence of the antagonistic action between strains of bacteria Bacillus subtilis VKPM B-10999, Rhodococcus erythropolis Ac-1882 and Pseudomonas putida In PMBC-10997 drug is determined by the growth of the strains in the places of contact of strokes on the environment and the method of agar blocks.

On LB agar medium cause a stroke .subtilis VKPM B-10999 and perpendicular to it, the strokes strains of Rhodococcus erythropolis Ac-1882 and Pseudomonas putida VKPM B-10997.

On the next Cup put a bar of Rhodococcus erythropolis Ac-1882 on nutrient medium and perpendicular to it, the strains of Bacillus subtilis VKPM B-10999 and Pseudomonas putida In PMBC-10997.

In the third bowl put the bar Pseudomonas putida VKPM B-10997 on Wednesday and perpendicular to it, the strains of Bacillus subtilis VKPM B-10999 and Rhodococcus erythropolis Ac-1882.

At the contact points of the strokes and the inhibition of growth of any of the strains do not see.

When the experiment by the method of agar blocks on the lawn of one of the strains on LB medium display units 3-day-old cultures of the two other strains.

In none of these experiments inhibit the growth of strains also do not mark that indicates the absence between antagonistic interactions and the possibility of their effective use.

The method of obtaining extracts of compost, vermicompost standard: soaking of vermicompost or compost in water and subsequent separation of the liquid fraction. JV the property allows you to obtain an extract, containing water-soluble substances (nutrients micro - and macronutrients and some physiologically active substances), as well as disputes soil bacteria, which when introduced into the soil multiply and enrich the soil necessary for the growth and development of plants, substances.

In a vessel with a capacity of 250 l put it in 10 kg of compost with a moisture content of 45%. Then it is soaked 3-5 volumes of water (30-50 l) at a temperature of 30-35°C. the mixture is stirred and stand for 30 min, receiving water bacterial extract containing water-soluble substances of the vermicompost and suspension of soil microorganisms - microbiocenosis of compost. Vermicompost usually includes the following trace elements: Cd, Co, Cr, Cu, Mo, Ni, Pb, Se, Zn.

When this mineral salts or mineral fertilizers, which are used as additives, are standard, commonly used when the soil or the soil to improve its quality. Use of nitrogen, phosphate, potash and mineral salt, such as Na2HPO4; KH2PO4; NaCl; NH4Cl, KCl, (NH4)2HPO4.

Examples of compositions of biological Fenox.

Example 3.

The biological Fenox, the form of dry powder

Dry biomass of cells (Konzentrat) - Bacillus subtilis VKPM B-10999 not less than 5×109CFU/g, Pseudomonas putida VKPM B-10997 not less than 5×109CFU/g and Rhodococcus erythroolis Ac-1882 at least 5×10 8CFU/g ratio (1-2):1:(1-2) (mass fraction - a 76.5-90%);

Without introduction or the introduction of the sorbent - little peat - 1-10% and/or promote supplements containing mineral salts and trace elements - vermicompost - 0,1-1%;

Water, not more than 10.0%;

The total titer of living cells is not less than 1010CFU/g

Example 4.

The biological Fenox, the form of dry powder

Dry biomass of cells of Bacillus subtilis VKPM B-10999, Pseudomonas putida VKPM B-10997 and Rhodococcus erythropolis Ac-1882 in the ratio of 1:1:1 - 5-30%;

Sorbent - little peat - 1-10% and/or

stimulating Supplement containing mineral salts and trace elements - vermicompost - 0,1-1%;

Water, not more than 10.0%;

The sorbent and the filler kaolin - up to 100%;

The total titer of living cells is not less than 5×108CFU/g

Example 5.

The biological Fenox, the form of dry powder

Dry biomass of cells of Bacillus subtilis VKPM B-10999, Rhodococcus erythropolis Ac-1882 and Pseudomonas putida In PMBC-10997 in the ratio of 1:1:1 in an amount of 5-30%;

Water, not more than 10.0%;

Kaolin-up to 100%;

The total titer of living cells is not less than 5×108CFU/g

Example 6.

The biological Fenox, the liquid form

The cells of the bacteria Bacillus subtilis VKPM B-10999, Rhodococcus erythropolis Ac-1882 and Pseudomonas putida VKPM B-10997 in the ratio of 1:1:1 (General title - 109CFU/ml) in the culture fluid without additives or with the addition of biologically active substances (extract of compost or in the of recomposit - 1-5 ml/l, or little compost or vermicompost 0.1% or/and chitin or chitosan, or hainstrasse raw - 0,05-0,4%).

Store at 0-8°C for at least 3 months, at temperatures of 18-25°C to 15 days.

When stored at temperatures of 18-25°C. Pets reduction in the number of viable cells to a titer FEW 107/ml.

The method of application is illustrated by the following examples.

Example 7.

The use of the drug Fenox for water purification from Imidacloprid

Drug Fenox in the form of a dry powder of the following composition: concentrate bacterial cells at a ratio of 1:2:1 with a total titer of 5×1010CFU/g (drug composition according to example 1) in an amount of 100 mg make 200 ml of tap water contaminated with the pesticide (insecticide) Imidacloprid at concentrations of 1 mg/l and 2 mg/l of active ingredient. Additionally in the aquatic environment contribute Supplement of mineral salts (g/l): Na2HPO4·7H2O - 6,4; KH2PO4- 1,5; 0.25 g/l NaCl; 0.5 g/l NH4Cl.

Control serve water samples contaminated with Imidacloprid at concentrations of 1 mg/l and 2 mg/l, with mineral salts without inoculants. Samples incubated at room temperature with constant mixing on a rocking chair (120 rpm). The repetition in experiment 3 times.

The content of Imidacloprid in water is determined standard chromatographic methods 1, 3, 5, 7 and 10-th day is and (Rudakov and others, 2004). The coefficient of variation of the data is not more than 5%.

Table 1
Dynamics of the concentration of Imidacloprid in water using the drug Phenox
OptionThe content of Imidacloprid, mg/l
0 days1 day3 dayday 710 days
Control - Imidacloprid 1 mg/ml1,000,980,980,940,96
Control - Imidacloprid 2 mg/ml2,002,092,011,931,95
Greentech + Imidacloprid 1 mg/ml1,000,940,810,480,28
Greentech + Imidacloprid 2 mg/ml2,00,57 1,150,830,64

The use of a biological product leads to a significant decrease in the content Imidacloprid in contaminated water. The decrease in the concentration of the pesticide in the original containerevent water Imidacloprid 1 mg/l for 10 days is 0.28 mg/l, less than 72%. When water contamination at a dose of 2 mg/l the introduction of a biological product leads to a decrease in the concentration Imidacloprid to 0.64 mg/l, reduced by 68%.

These data indicate that the biological Fenox allows cleaning of water from Imidacloprid.

Example 8.

The use of biological Fenox for water purification from phenol

The biological Fenox in the form of a liquid form of the following composition: bacterial cells at a ratio of 1:1:1 with a titer of 5×109CFU/g (drug composition according to example 1 without the addition of stimulus components) in an amount of 0.01% of the volume make contaminated with phenol water (the content of phenols 100 mg/l). Additionally in the aquatic environment contribute mineral nitrogen-phosphate fertilizer in the amount of 0.5 g/l For aeration and mixing in the tank with contaminated water periodically, the air is supplied by a pump. The temperature of the water supported by about 22-25°C. the Determination of phenol in water conduct standard photometric method (Rudakov and others, 004). The repetition in the experiments 3 times. The coefficient of variation of the data is not more than 7%.

Table 2
Dynamics of the content of phenol in water treatment with the drug Fenox
The incubation periodThe concentration of phenol, mg/l
0 days100,0
1 day73,0
3 day67,0

When using the drug Fenox concentration of phenol in water for the first day is reduced by 27%, and by 3 days-33% (table 2).

Example 9.

The use of the drug Fenox for water purification from dichlorphenol

The experience carried out according to example 8, but using the ratio of strains 2:1:1 and water contaminated with dichlorophenol 2,4-DHF (content dichlorphenol 100 mg/l).

Table 3
Dynamics of the content of dichlorophenol in water treatment with the drug Fenox
The incubation periodThe concentration of 2,4-DHF, mg/l
0 days100,0
1 day30,0
3 dayof 21.9

When use of the drug Fenox for water purification from dichlorphenol concentration of 2,4-DHF is reduced to 1-day incubation at 70% (table 3).

Example 10.

The use of the drug Fenox for water purification from 4-chlorophenoxyacetic acid

The experience carried out according to example 8, but using the ratio of strains 1:2:1 and water contaminated with 4-chlorophenoxyacetic acid 4-HFUC (contents 4-HFUC 100 mg/l).

Table 4
Dynamics of the concentration of 4-chlorophenoxyacetic acid in water treatment with the drug Fenox
The incubation periodThe concentration of 4-HFUC, mg/l
0 days100,0
1 day96,3
3 day95,8
5 day94,1
day 792,5
9 suck the 90,0
11 day68,7
13 day63,9
day 1562,0

The use of the drug Fenox for water purification from 4-chlorophenoxyacetic acid to 9-th day reduces the concentration of 4-HFUC in water at 10%, and 15 m day - by 38.0% (table 4).

Example 11.

The use of the drug Fenox for water purification of 2,4-dichlorophenoxyacetic acid

The experience carried out according to example 8, but using water contaminated with 2,4-dichlorophenoxyacetic acid 2,4-D (the content of 2,4-D 100 mg/l).

Table 5
Dynamics of the concentration of 2,4-dichlorophenoxyacetic acid in water treatment with the drug Fenox
The incubation periodThe concentration of 2,4-D, mg/l
0 days100,0
1 dayfor 95.3
3 day89,4
5 day81,5
day 7 74,3
9 day70,0
11 day68,6
13 day66,9
day 1565,0

When using the drug Fenox for water purification of 2,4-dichlorophenoxyacetic acid content of 2,4-D gradually falls to 3-day by 11%, to 9-day - 30% and to 15-day - 35%.

Example 12.

The use of the drug Fenox for water purification from 2,4,5-trichlorophenoxyacetic acid

The experience carried out according to example 8, but using water contaminated 2,4,5-trichlorophenoxyacetic acid 2,4,5-T (the content of 2,4,5-T 100 mg/l).

Table 6
Dynamics of the concentration of 2,4,5-trichlorophenoxyacetic acid in water treatment with the drug Fenox
The incubation periodThe concentration of 2,4-DHF, mg/l
0 days100,0
1 day96,3
3 day85,0
5 day64,8
8 days55,0

When using the drug Fenox for water purification from 2,4,5-trichlorophenoxyacetic acid concentration of 2,4,5-T for 3 days reduced by 15%, to 8-day - on 45% from the initial level (table 6).

Example 13.

The use of the drug Fenox for water purification from phenol

The experience carried out according to example 12, but use of the drug by Fenox in the form of a dry powder (example 3, with a titer of 5×1011CFU/g and the introduction of vermicompost - 0,1%).

Table 7
Dynamics of the content of phenol in water treatment with the drug Fenox
The incubation periodThe concentration of phenol, mg/l
0 days100,0
1 day70,0
3 day45,0

When use of the drug Fenox for in the form of a dry powder with catalytic additive for water purification from phenol its content decreases towards the end of 1 day to 30%by the end of 3 days at 55% of the initial level (table 7).

Example 14.

the drug Fenox for water purification of 2,4-dichlorophenoxyacetic acid

The experience carried out according to example 13, but using the product from example 3, without vermicompost with title 1011CFU/g and water contaminated with 2,4-dichlorophenoxyacetic acid 2,4-D (the content of 2,4-D 100 mg/l).

Table 8
Dynamics of the concentration of 2,4-dichlorophenoxyacetic acid in water treatment with the drug Fenox
The incubation periodThe concentration of 2,4-D, mg/l
0 days100,0
2 day81,3
4 days64,2
6 days61,8
8 daysof 60.5
10 days59,0

The use of the drug Fenox for water purification of 2,4-dichlorophenoxyacetic acid leads to gradual decrease in the concentration of 2,4-D, on the 10th day she falls to 59% from the original values (table 8).

Example 15.

The use of the drug Fenox for water purification from dichlorphenol

The experience carried out according to example 13, but using water, Zahra is United by dichlorophenol 2,4-DHF (content dichlorphenol 100 mg/l).

Table 9
Dynamics of the content of dichlorophenol in water treatment with the drug Fenox
The incubation periodConcentration
2,4-DHF, mg/l
0 days100,0
2 day96,2
4 days91,4
6 days86,9
8 days80,5
10 days75,7
12 day70,1
14 day64,9
16 day60,8
18 day58,7
20 days56,6
22 day53,0

When use of the drug Fenox for water purification from dichlorphenol concentration is the situation 2,4-DHF is to 22 days 53% of the initial (table).

Example 16.

The use of the drug Fenox for water purification from 2,4,5-trichlorophenoxyacetic acid

The experience carried out according to example 8, but using the drug with stimulant additives and title 109CFU/g and water contaminated 2,4,5-trichlorophenoxyacetic acid 2,4,5-T (the content of 2,4,5-T 100 mg/l).

Table 10
Dynamics of the concentration of 2,4,5-trichlorophenoxyacetic acid in water treatment with the drug Fenox
The incubation periodThe concentration of 2,4,5-T, mg/l
0 days100,0
2 day70,0
4 days35,2
6 daysto 33.8
8 days33,1
10 days32,6
12 day31,0
14 day31,9
16 day38,7
18 the ducks 35,8
20 days28,6
22 day30,1
24 daysof 31.4
26 day18,3
28 days16,0

The content of 2,4,5-T in water consistently reduced after application of the drug Fenox - to 2-day 30%, and further to 12-m - 69%, and 28-day - 84% (table 10).

Example 17.

The use of biological Fenox for industrial wastewater purification from phenol

The biological Fenox in the form of a dry powder of the following composition: dry biomass of cells of Bacillus subtilis VKPM B-10999, Rhodococcus erythropolis Ac-1882 and Pseudomonas putida In PMBC-10997 in the ratio of 1:1:1 in the amount of 10%; vermicompost - 1%, water, not more than 10.0%, kaolin up to 100%; the titer of living cells is 2×109CFU/g (prepared according to example 4).

Before applying in a container (bucket, jar) dry powder product is well mixed in warm water (25-30°C) until a homogeneous suspension.

In a tank with a working volume equipped with a stirrer or device for bubbling air, contribute to 1.0 kg of a biological product, 0.5 kg of Dimovski (NPK) and add 200 l of hot water. To activate mikroorganizmov.pri mixed or blown with air for 24 hours. Then prepare a working suspension of the drug, add it to the tank with waste water volume of 10 m3drains, 10 kg of mineral fertilizers (NPK) and 1 kg of lime. The tank sewage daily purge air.

Wastewater pollution JSC "ufahimprom" phenolic compounds is 30.4 mg/L. the Efficiency of wastewater treatment from phenols by biological Fenox appreciate without additives and with the addition of mineral fertilizers in wastewater (table).

Table 11
The dynamics of industrial wastewater purification from phenol in the application of biological Fenox
The period after the introduction of strain in the effluent of the biological Fenox, d
OptionThe content of phenol, mg/lThe efficiency of wastewater treatment from phenols, %
02570257
Sinks 30,410,20,010,01066,40099,97099,970
The effluent with the addition of mineral salts30,43,20,0010,001089,500of 99.997of 99.997

The degree of wastewater purification from phenol on the 2nd day after making biological Fenox reaches 66,4%, and by the 5th day is more than 99%. The efficiency of biological Fenox for removal of phenols increases when added to the effluent of mineral salts, by the 2nd day the degree of wastewater purification from phenol reaches to 89.5%and to 7 days - wastewater freed from phenol by more than of 99.997% (table).

Example 18.

The use of biological Fenox for industrial wastewater purification from phenol.

The industrial wastewater treatment carried out according to the previous example without the addition of mineral fertilizers, but use drains LLC "Novo-Ufimsky oil refinery", JSC "tanning agent".

Table 12
The dynamics of industrial wastewater purification from phenol in the application of the strain of biological Fenox
OptionPeriod after making the strain of biological Fenox drains, d
The content of phenol, mg/lThe efficiency of wastewater treatment
from phenols, %
01370137
Drains petrochemical enterprises0,0960,0640,0120,012066,787,587,5
The effluents of the leather industry0,750-0,1550,1550- 79,479,4

The degree of wastewater purification petrochemical enterprises from phenols on the 1st day after making biological Fenox amounts to 66.7%and 3-7-day reaches of 87.5%. The efficiency of biological Fenox for treatment of phenol wastewater tanning production is to 3-7 days 79.4 per cent (table).

Example 19.

The use of the drug Fenox for the destruction of oil in the aquatic environment

Medication in the form of a dry powder (composition according to example 3) with a titer of 1×109CFU/g in the amount of 100 mg make 200 ml of water contaminated with crude oil (concentration 1%) with the addition of salts (K2HPO4- a 0.05%, NH4Cl - 0,05%, caso3- 0,001%). Contaminated oil-water after inoculation of the drug and mineral salts of nitrogen, phosphorus, potassium and lime incubated for 10 days at room temperature under static conditions, with daily stirring. Control is an option without making the drug. The repetition in experiment 3 times.

On the surface of the water in the variant with the drug for 3 days to celebrate the dark flakes in water volume and smaller spots of oil on the surface than in the control. On the 5th day in the variant with drug see a small amount of oil spots on the surface of the water, about half-destroyed the oil film on the flask walls. The 10-day experience mark is given a small amount of oil on the water surface and the vessel wall.

The mass concentration of oil in the samples was measured on the 10th day concentrator KN-2 (manufacturing LLC NEP CIBACOPA, Novosibirsk) spectrophotometric method for the level of electoral oil absorption in the infrared region of the spectrum at wavelength 3,42 micrometer [Methods of measurement of mass concentration of NP in samples of drinking water and wastewater by the method of infrared spectrophotometry PND f 14.1:2:4.168-2000, Novosibirsk, 1998, 17 C.]. The coefficient of variation of the data is not more than 5%.

The drug reduces the oil in water content of 28% (2652 mg/l in the control to 1915 mg/ml in a variant of the biological product).

Example 20.

Survival of bacterial strains of the drug in the soil

The population dynamics of strains of bacteria of the drug in the soil studied in model experiments. Cell suspension in an amount of 3×108cells in 1 g of soil contribute in glass vials with 50 grams of leached Chernozem. The experiment included 3 options: without xenobiotic (control) and 2,4-D at concentrations of 100 and 10000 MAC. MPC for 2.4-D in soil is 0.1 mg/kg soil Humidity is 60% of full capacity. Periodically conduct the selection of strains of drug Fenox on minimal salt agar medium M9 with the addition of 2.4-D as the sole source of carbon and energy.

The number of colony forming units (the SECOND) strains of bacteria in the control soil increases for 30 days, and then see a decline in the population density of these bacteria (table 13). In contaminated 2,4-D population density of bacteria is higher during the first 2.5 weeks after making than in the control, and then gradually decreases. In the most contaminated soil, the number of bacteria Finaxa sharply increases in 10-50 times the population density of this strain in control. Such a rise in the number of bacteria Finaxa due to the use of xenobiotic as a power source. Reducing the number of bacteria Finaxa in the soil of this variant occurs after 30 days. To 52 days the population density of the introduced strains at the same level in all variants, which indicates a reduction in contaminated soils the concentration of 2,4-D, as well as possible involvement in the degradation of pesticide and other microorganisms.

Identified good adaptation of microorganisms to the soil. Similar data are obtained in experiments with soils of other types of grey forest, podzolic soil and xenobiotic in three concentrations of 100 MPC, MPC 1000, 10000 MAC. MPC for 2.4-D in soil is 0.1 mg/kg

The principal possibility of the use of the drug of the present invention in these soil types.

Table 13
Dynamics Chi the industry strains of drug Fenox when applied to contaminated 2,4-D pesticide leached Chernozem
PeriodPopulation dynamics of a population of cells, SOME
time×105/g soil
Clean soilSoil with 2,4-D 100 MPCSoil with 2,4-D MPC 1000Soil with 2,4-D 10000 MAC
0 days0,000,000,000,00
9 days0,374,334,330,66
19 days13,4050,504,30700,00
30 days70,0055,409,00340,50
52 days1,703,002,700,98

Example 21.

The use of biological Fenox for cleaning soil from pesticide 2,4,5-T (rechlorination acid)

Soil (100 g) of the arable horizon sieved through a sieve of 2 mm diameter and placed in glass vials for 500 ml soil making 2,4,5-T in the amount of 100 mg/g and mix thoroughly.

The biological Fenox with title 109CFU/g (according to example 5) inoculant in a bottle with soil, moistened to 60% of full capacity and also mix. The bottles are closed with cotton plugs with parafilm and incubated at a constant humidity and a temperature of 25°C in a thermostat.

The definition of 2,4,5-T in soil is carried out by a standard method (link). The repetition in the experiments 3 times. The coefficient of variation of the data is not more than 8%.

Table 14
Dynamics of cleaning soil from 2,4,5-T in the application of biological Fenox
IndexTime after tillage, day.
151014213048
The content of 2,4,5-T in soil, mg/g10095,087,672,668,133,5
The degree of purification to control, %05,012,415,027,431,966,5

The degree of purification of soil from 2,4,5-T when using a strain of biological Fenox is in 14 days - 15%, 21 day - 27.4 per cent and 30 days - 31.9% and reaches to 48 day - to 66.5%. The rate of decomposition of 2,4,5-T increases significantly after 30 days of incubation of soil with biological Fenox (table 14).

Example 22.

The use of the drug Fenox for the decomposition of the pesticide (fungicide) TMTD in the soil

Venue experience: concrete Playground adjacent to the storage of pesticides (Altai territory, Biysk, Suburban street, 4).

In a summer filled with concrete on the outside of the space suit plot size of 3 m2(1×3 m), separated by a wooden partition with a height of 20 cm 3 plot (frequency) of size 1×1 m Bottom plots spread plastic wrap to prevent the spread of pollutants from the soil into concrete pad. The plot is filled with a layer of black soil thickness of 15 cm At a height of 60 cm from ground plots close nonwoven Ukr the main material (spunbond). In the soil contribute pesticide (fungicide) tetramethylthiuramdisulphide (TMTD) at the rate of 50 g/m2the vermicompost in the amount of 50 g/m2mineral nitrogen-phosphate fertilizer enriched with trace elements, - 100 g/m2and organic fertilizer on the basis of peat - 400 g/m2and mix thoroughly.

Drug Fenox in the form of a dry powder of the following composition: bacterial cells - 10%, vermicompost - 1%, kaolin - 89% with titer of 5×109CFU/g (according to example 4) in an amount of 300 g was dissolved in 15 liters of water (25-30°C) and incubated for 24 hours with periodic mixing to improve aeration and activated microorganisms. Suspension of a biological product applied to the surface of the plots at a rate of 5 l/m2by watering with a watering can. Surface plots additional water in an amount of 5 l/m2soil.

Soil, contaminated TMTD, after spraying Phenox loosened to improve aeration and implement the mulch with a thin layer of sawdust. If necessary, and the lack of rainfall the soil on the plot periodically loosened and moisturized up to 60-70% of full capacity. To assess the effectiveness of cleaning soil from TMTD method of averaged samples selected samples after 1.5 and 3 months. Quantitative content TMTD in samples determined chromatographically. The coefficient of variation of the data is not more than 5%.

1.5 months after spraying Phenox content TMTD in the soil is reduced by 53%, after 3 months at 75%.

Example 23.

The use of the drug Fenox for the decomposition of the pesticide (herbicide) dichloramine in the soil

The experience carried out analogously to example 22. In the soil contribute pesticide (herbicide) dichloramine at the rate of 50 g/m2.

1.5 months after spraying Phenox content TMTD in the soil is reduced by 59%, after 3 months, 78%.

Example 24.

The use of the drug FENOX (Phenox) for cleaning soil from chloroaromatics connections

Venue experience: Rostov region, Salsk district.

Characteristics of plot: agricultural land, polluted chloroaromatics connections.

In the summer on the open ground lay out the plot area of 530 m2(size 66 m × 8 m)are indicated on the plots with an area of 170 m2protective strips of 0.5 m Repetition in experiment 3 times.

In contaminated soil making vermicompost in the amount of 100 g/m2mineral nitrogen-phosphate fertilizer at the rate of 10 g/m2.

Drug Fenox (Phenox) in the form of a dry powder of the following composition: bacterial cells - 10%, vermicompost - 1%, kaolin - 89% with titer of 5×109CFU/g (according to example 4) in an amount of 10 g/m2to obtain a working solution dissolve the container (bucket, the flask and mixed well in warm water (25-30°C) until a homogeneous suspension.

A working suspension of the drug Phenox applied to the soil surface at the rate of 10 l/ m Application is carried out by sprinkling with any dedicated machines and aggregates. Treatment is carried out at a daily average soil temperatures not below +5°C and not above +30°C.

Made biologic and fertilizer is worked into the top contaminated soil by ploughing, perekapyvanie, cultivation or other techniques groovebot.

The soil periodically loosened to improve aeration and moisturize up to 60-70% of full capacity (if necessary, the lack of rainfall). The soil at the site two weeks replowed and moisturize.

To assess the effectiveness of cleaning soil from chloroaromatics compounds by the method of averaged samples selected samples after 1.5 and 3 months. Quantitative content chloroaromatics compounds in the samples determined with the standard method (Rudakov and others, 2004). The coefficient of variation of the data is not more than 9%.

1.5 months after treatment with the drug FENOX (Phenox) content chloroaromatics compounds in the soil is reduced by 35%in 3 months - 68%.

Example 25.

Evaluation of fungicidal activity of the drug.

Antifungal activity of strains of bacteria .subtilis VKPM B-10999, Rhodococcus erythropolis Ac-1882 and Pseudomonas utida VKPM B-10997 drug determine the standard method agar blocks (Egorov, 1976).

A Petri dish with solid nutrient medium inoculated with a lawn of phytopathogenic fungal culture. Use strains of fungi, causing damage roots, decay, blight, root rot - Fusarium oxysporum, Alternaria tennuisima, Alternaria alternata and grey mould on leaves and fruits of plants, Botrytis cinerea, rot agricultural products - Geotrichum candidum.

Every culture seeded 2 cups with the environment of čapek and on the surface of the display blocks bacterial strains, carved with a 5-day lawn of bacteria on standard LB-agar. Display 4 block bacterial strain on every Wednesday, planted by phytopathogenic fungus. Cup incubated for 2 days at 5°C (for the diffusion of antibiotic substances in the agar, and then at 25°C and establish the presence of zones of inhibition of growth of fungi.

Table 15
Fungicidal activity of bacterial strains medication
The bacterium
tion strain
The size of the zone of inhibition of the fungus on the 3rd day, mm
Fusarium oxysporumAlternaria alternataAlternaria tennuisimaBotrytis cinereaGeotrichum candidum
Bacillus subtilis13-*1-
Rhodococcus erythropolis12-22
Pseudomonas putida12232
* no zone suppression

Fungicidal activity against all tested in the experience of phytopathogenic fungi establish the strain Pseudomonas putida VKPM B-10997 and strains .subtilis VKPM B-10999, Rhodococcus erythropolis Ac-1882 to the fungi that cause predominantly root infection and rot (table). That is, bacterial drug has fungicide activity against the pathogens causing various fungal diseases of plants.

Stimulating plant growth activity of the drug is

Example 26.

The use of drugs to stimulate the growth of radish plants (DRUG?)

Medication in the form of a dry powder of the following composition: bacterial cells - 10%, vermicompost - 1%, kaolin - 89% with titer of 5×109CFU/g of restore the t in 10 ml of distilled water (1:10 dilution) and then prepare serial dilutions 1:100 and 1:1000. Seeds of radish varieties of Pink-red with a white tip laid on filter paper in Petri dishes at 25 pieces per Petri dish. The repetition in experiment 4 times. Seeds treated with the suspension of the drug in the amount of 2 ml per Petri dish using an automatic dispenser. Control options are seeds germinated in distilled water, and seeds treated with 0.05% solution indolylmethane acid (IBA) is a stimulator of plant growth auxinic type. Petri dishes with treated seeds incubated under natural light and room temperature in a humid chamber.

The length of roots, length of shoots and dry weight of plants was determined on 5 days. The coefficient of variation of the data is not more than 5%.

Table 16
The growth of radish plants if the drug is
Treatment optionIndex
root length, mmthe length of the sprout, mmdry weight (air-
dry weight, g
Control (water)69300,27
Control (IMC)70370,35
Drug dilution of 1:1061410,34
Drug dilution of 1:10081420,37
Drug dilution of 1:100058380,32

Suspension of the drug stimulates the growth of radish red and biomass accumulation. The maximum effect observed from the aqueous suspension of the drug 1:100. Drug dilution of 1:100 increases the length of the roots of 5-day radish from 69 to 81 mm (17%), length of seedlings from 30 to 42 mm (40%), and dry weight of plants by 37%, from 0.27 to 0.37 g (table 16).

Example 27.

The use of drugs to stimulate the growth of plants cabbage

The experience carried out analogously to example 26. The dilutions of the drug treated seeds cabbage varieties Glory. The length of roots and shoots of plants determined after 5 days.

Table 17
The growth of cabbage varieties Glory if the drug is
Treatment optionIndex
root length, mmthe length of the sprout, mm
Control(water)4631
Control (IMC)5236
Drug dilution of 1:106435
Drug dilution of 1:1006937
Drug dilution of 1:10006337

Aqueous suspension of the drug (in all tested dilutions) increases the length of the root to 50% from 46 to 63-69 mm and a length of seedlings by 19% - from 31 to 37 mm at a dilution of 1:100 and 1:1000 (table 17). These findings indicate a strong stimulating effect on root growth of cabbage varieties Glory.

Example 28.

The use of drugs to stimulate seed germination and plant growth of clover

The experience carried out analogously to example 26. R is svedeniya drug treated seeds red clover. After 5 days determine the length and root length of seedlings, germination and dry weight of plants.

Table 18
The growth of red clover if the drug is
Treatment optionIndex
root length, mmthe length of the sprout, mmgermination, %dry weight, g
Control(water)1830880,07
Control (IMC)2333910,09
Drug dilution of 1:103440840,12
Drug dilution of 1:10024331000,15
Drug dilution of 1:1000 2536960,12

The drug in 1:10 dilution increases the linear sizes of plants - the length of the root by 89% (from 18 to 34 mm) and the length of the sprout by 33% (from 30 to 40 mm). Drug dilution of 1:100 promotes germination by 14% from 88 to 100% and the accumulation of dry matter by 114% from 0.07 to 0.15 g (table). So, the drug has a strong stimulating effect on root growth and accumulation of mass of plants clover.

Example 29.

The use of drugs to stimulate the growth of cucumber plants

The experience carried out analogously to example 26. The dilutions of the drug treated seeds of cucumber cultivar Russian shirt F1. The length of roots and seedlings, germination and dry weight of plants determined after 6 days.

33
Table 19
The growth of cucumber plants if the drug is
Treatment optionIndex
root length, mmthe length of the sprout, mmgermination, %dry weight, g
Control(water)46840,46
Control (IMC)5938920,55
Drug dilution of 1:107441960,72
Drug dilution of 1:10071411000,70
Drug dilution of 1:10006436920,63

Aqueous suspension of the drug have a stimulating effect on the growth of cucumber cultivar Russian shirt F1. For example, in water 1:10 dilution of the drug increases the length of the root by 61% from 46 to 74 mm, length of sprout 24% - 33 to 41 mm, the germination rate by 14% from 84% to 96% and dry weight of plants at 37% and 0.46 to 0.63 g (table 19). The drug is the most powerful effect is on the root growth of cucumber.

Example 30.

The use of drugs to stimulate germination of beet

The experience carried out analogously to example 26. The dilutions of the drug treated seeds beet varieties Detroit. The germination of Radelet on day 7.

Table 20
The germination of beet drug use
Treatment optionIndex
germination, %
Control (water)92
Control (IMC)95
Drug dilution of 1:1096
Drug dilution of 1:10099
Drug dilution of 1:100096

Drug dilution of 1:100 promotes seed germination of beet by 8% from 92 to 99% (table).

Example 31.

The use of drugs to stimulate plant growth onions

The experience carried out analogously to example 26. Drug in dilutions 1:100 and 1:1000 treated seeds onion variety Carmen. The length of roots and seedlings, germination and dry weight of plants determine within 10 days.

Table 21
The growth of plants when Luke application : the preparation
Treatment optionIndex
root length, mmthe length of the sprout, mmdry weight, g
Control (water)30570,19
Control (IMC)33610,21
Drug dilution of 1:10034690,28
Drug dilution of 1:100037620,25

Drug dilution of 1:1000 increases the length of the root by 23% - from 30 to 37 mm, the length of the sprout 9% from 57 to 62 mm and the accumulation of plant dry weight by 32% - from 0.19 to 0.25 g (table 21).

Example 32.

The use of drugs to stimulate plant growth carrots

The experience carried out analogously to example 31. The dilutions of the drug treated seeds carrot cultivar Red giant. Within 10 days to determine dry weight of plants.

Table 2
Treatment optionIndex
dry weight, g
Control (water)0,03
Control (IMC)0,05
Fenox+BL-5 dilution of 1:100,04
Fenox+BL-5 dilution of 1:1000,04
Fenox+BL-5 dilution of 1:10000,05

Drug dilution of 1:1000 increases dry weight of seedlings carrots by 67%, from 0.03 to 0.05 g (table 22).

The presence of pesticides in soil, water and industrial effluents was carried out using gasochromatic method for determination of pesticides described Rudakov O., and others (the Companion of chromatographica. - Voronezh: Aquarius, 2004. - 528 S.).

1. Biological product for cleaning of ground, soil, industrial waste water from decay-resistant pesticides, selected from chlorophenoxyacetic acids -2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T), chlorprothixene acid (HPUK), venexiana acid (FOOK), -2,4-dichlorophenoxy-α - propionic acid, 2-methyl-4-chlorophenoxy-α-propionic acid, 2,4,5-trichlorophenoxy-α-propionic acid, 2,4-Dich is orphenochs-α-butyric acid, methyl-[1-(butylamino)carbonyl]-1H-benzimidazole-2-ylcarbamate, 2,4-dichlorphenol, Imidacloprid, hexachlorohexane, as well as phenol, representing the Association of bacterial strains Pseudomonas putida VKPM B-10997, Bacillus cubtilis VKPM B-10999, u Rhodococcus erythropolis PMBC-Ac-1882 mass ratio of (1-2):(1-2):1.

2. Biological preparation according to claim 1, additionally containing sorbent and possible mineral, organic and stimulating supplements.

3. Biological preparation according to claim 1 or 2 to clean the ground, soil, industrial waste water from decay-resistant pesticides, selected from chlorophenoxyacetic acids, such as -2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T), chlorophenoxyacetic acid (HPUK), venexiana acid (FOOK), and 2,4-dichlorophenol and phenol, Imidacloprid.

4. Biological preparation according to claim 1, additionally possessing plant growth and fungicidal action.

5. The method of use of a biological product for cleaning of ground, soil, industrial waste water from decay-resistant pesticides, selected from chlorophenoxyacetic acids -2,4-dichlorophenoxyacetic acid (2,4-D), trichlorophenoxyacetic acid (2,4,5-T), chlorprothixene acid (HPUK), venexiana acid (FOOK), -2,4-dichlorophenoxy-α-propionic acid, 2-methyl-4-chlorophenoxy-α-propionic acid, 2,4,5-trichlorophenoxy-α-propionic acid, 2,4-dichlorophenoxy-α-wt is Jana acid, methyl-[1-(butylamino)carbonyl]-1H-benzimidazole-2-ylcarbamate, 2,4-dichlorphenol, Imidacloprid, hexachlorohexane, as well as phenol, the introduction of contaminated soil, soil or industrial runoff preparation according to any one of claims 1 or 2 in an effective amount.

6. The method according to claim 4, characterized in that the drug is administered in an amount of 10 kg of dry matter at the time of ploughing based on 1 ha of land.

7. The method according to claim 4, characterized in that the drug is administered in the form of an aqueous solution by sprinkling.



 

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1 dwg, 3 ex

FIELD: biotechnologies.

SUBSTANCE: invention relates to oligonucleotide primers and the method of their use for detection of Lactobacillus delbrueckii subspecies bulgaricus in starter cultures. Proposed synthetic oligonucleotide primers have nucleotide sequences: Lbul4F 5'-GGCCAGCCAGATCGCCAGC-3' and Lbul5R 5'-GACCAGGTCGCTGTCCGGC-3'. The proposed method includes performance of PCR. In case a DNA ferment is detected with size of 409 base pairs. a conclusion is made on availability of Lactobacillus delbrueckii subspecies bulgaricus in the investigated biomaterial.

EFFECT: inventions may be used in milk processing industry for detection and identification of strains and cultures Lactobacillus delbrueckii subspecies bulgaricus in starter cultures used in production of cultured milk foods.

2 cl, 1 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: nutrient medium contains HMF - agar, erythrocytic mass from donor's human blood, serum of cattle and yeast extract at the specified ratio.

EFFECT: invention makes it possible to increase sensitivity of medium to extracted microorganisms, to improve growth properties of nutrient medium and to simplify method of its preparation.

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry and use of a Trichoderma harzianum Rifai strain as a producer of an Impatiens necrotic spot tospovirus inhibitor. The Trichoderma harzianum Rifai strain, which is deposited in the Russian National Collection of Industrial Microorganisms under No.F-180, is a producer of the homogeneous enzyme L-lysine-a-oxidase, which exhibits marked antiviral activity with respect to Impatiens necrotic spot tospovirus.

EFFECT: reduced loss of decorative and vegetable crops caused by Impatiens necrotic spot tospovirus.

2 ex

FIELD: biotechnologies.

SUBSTANCE: strain of bacterial Pseudomonas panipatensis VKPM V-10593 is proposed, which is capable of quick recycling of oil and oil products, in particular, diesel fuel. Strain may be used to clean soil and water reservoirs contaminated with oil and oil products, in a wide range of temperatures from +8 to +30°C.

EFFECT: improved properties of a strain.

2 tbl, 5 ex

FIELD: biotechnologies.

SUBSTANCE: method provides for degreasing of buckwheat straw with 100% acetone. The degreased straw is extracted in 5 l of 70% ethyl alcohol in boiling water bath for 1 hour. The extracted raw materials are separated by filtration to produce alcohol extract. The produced alcohol extract of the stimulant is concentrated under vacuum, cleaned from admixtures with ethyl acetate and dried to prepare target product.

EFFECT: invention makes it possible to increase yield of yeast biomass.

2 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention relates to biotechnology, and represents a method for preparing L-arginine with the use of the bacteria of to the genus Escherichia, which is modified in such a way that the astCADBE operon in the mentioned bacterium is inactivated. The bacterium is grown in a culture medium, after that L-arginine is recovered from a culture fluid.

EFFECT: method enables providing higher productivity of the bacteria producing L-arginine and producing high-yield L-arginine.

4 cl, 2 tbl, 14 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention relates to biotechnology, and represents a method for preparing L-arginine with the use of the bacteria of to the genus Escherichia, which is modified in such a way that the astCADBE operon in the mentioned bacterium is inactivated. The bacterium is grown in a culture medium, after that L-arginine is recovered from a culture fluid.

EFFECT: method enables providing higher productivity of the bacteria producing L-arginine and producing high-yield L-arginine.

4 cl, 2 tbl, 14 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. The Bacillus atropheus IPNG - ELA-2 bacteria strain, having recycling capacity with respect to oil and oil products, is deposited in the Russian National Collection of Industrial Microorganisms (VKPM) under registration number VKPM V-10592 and can be used to clean water and soil from oil.

EFFECT: invention enables to cut the duration of recycling oil and oil products.

4 tbl, 6 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Obtained is a Lactobacillus rhamnosus 38k bacteria strain, having antagonistic activity with respect to pathogens and opportunistic pathogens, including Candida yeast fungi, average adhesion and self-aggregation properties, high surface hydrophobicity, average coaggregation, is deposited in the Russian National Collection of Industrial Microorganisms (VKPM) under registration number VKPM V11155.

EFFECT: strain can be used in production of probiotic bacterial preparations, biologically active food additives, fermented milk and non-fermented food products.

5 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and dairy industry. The method of producing a culture medium for culturing lactose-fermenting yeast involves the following: Tap water is mixed with Arshan resort mineral water with mineral content of 4.1 mg/l in ratio of 1:1, followed by addition of baking yeast in amount of 10-12 g/l of the mixture of tap water and mineral water and a yeast overcook is prepared. The obtained yeast overcook is filtered and sterilised and peptone and lactose are added in amount of 1% and 4%, respectively, per 100 ml of the yeast overcook, to obtain the medium. The obtained medium is re-sterilised and cooled.

EFFECT: invention increases biomass output of lactose-fermenting yeast.

1 dwg, 3 ex

FIELD: biotechnologies.

SUBSTANCE: invention relates to oligonucleotide primers and the method of their use for detection of Lactobacillus delbrueckii subspecies bulgaricus in starter cultures. Proposed synthetic oligonucleotide primers have nucleotide sequences: Lbul4F 5'-GGCCAGCCAGATCGCCAGC-3' and Lbul5R 5'-GACCAGGTCGCTGTCCGGC-3'. The proposed method includes performance of PCR. In case a DNA ferment is detected with size of 409 base pairs. a conclusion is made on availability of Lactobacillus delbrueckii subspecies bulgaricus in the investigated biomaterial.

EFFECT: inventions may be used in milk processing industry for detection and identification of strains and cultures Lactobacillus delbrueckii subspecies bulgaricus in starter cultures used in production of cultured milk foods.

2 cl, 1 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: nutrient medium contains HMF - agar, erythrocytic mass from donor's human blood, serum of cattle and yeast extract at the specified ratio.

EFFECT: invention makes it possible to increase sensitivity of medium to extracted microorganisms, to improve growth properties of nutrient medium and to simplify method of its preparation.

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry and use of a Trichoderma harzianum Rifai strain as a producer of an Impatiens necrotic spot tospovirus inhibitor. The Trichoderma harzianum Rifai strain, which is deposited in the Russian National Collection of Industrial Microorganisms under No.F-180, is a producer of the homogeneous enzyme L-lysine-a-oxidase, which exhibits marked antiviral activity with respect to Impatiens necrotic spot tospovirus.

EFFECT: reduced loss of decorative and vegetable crops caused by Impatiens necrotic spot tospovirus.

2 ex

FIELD: biotechnologies.

SUBSTANCE: strain of bacterial Pseudomonas panipatensis VKPM V-10593 is proposed, which is capable of quick recycling of oil and oil products, in particular, diesel fuel. Strain may be used to clean soil and water reservoirs contaminated with oil and oil products, in a wide range of temperatures from +8 to +30°C.

EFFECT: improved properties of a strain.

2 tbl, 5 ex

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