RU2697723C1 - Agent and method for decontaminating soil areas contaminated with anthrax or bacteria of agents of dangerous and especially dangerous infections - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: group of inventions refers to medicine, veterinary science, microbiology, namely to disinfectology technologies. Decontamination agent of soil areas contaminated with spores of bacteria of an anthrax agent or bacteria of agents of hazardous and especially dangerous infections involves medical hydrogen peroxide, alkyldimethylbenzylammonium chloride (ADBAC) and water. Preparation additionally contains calcium chloride and sulphanol, with the following content of components, wt%: medical hydrogen peroxide – 30.0±3.0; alkyldimethylbenzylammonium chloride – 2.0±0.2; calcium chloride – 1.0±0.1; sulphanol – 1.0±0.1, water – balance. Also disclosed is a method for decontaminating soil areas contaminated with spores of bacteria of anthrax agent or bacteria of agents of hazardous and especially dangerous infections by said agent.EFFECT: group of inventions provides higher efficiency of contaminated soil areas disinfection.2 cl, 3 tbl, 5 ex

Description

The invention relates to the field of medicine, veterinary medicine, microbiology, and in particular to disinfectological technologies and to ready-made chemical composite disinfectants designed to eliminate soil foci of dangerous and especially dangerous infections resulting from the burial, slaughter, mortality of sick animals or as a result of technological accidents and disasters or bioterrorist acts.

There is a method of disinfecting soil, mainly contaminated with the causative agent of anthrax, by introducing actinomycetes of the genus Cinereus (species Actiunomycesgriseus, A. griseovariabilis, A. chromophuseus) into the soil of anthagonists of anthrax (copyright certificate No. 528917, IPC A01N 7/04). The method is based on the use of actinomycetes of antibiotic producers as natural antagonists of the causative agent of anthrax.

The disadvantage of this method is its low efficiency, since actinomycetes and their metabolic products are partially effective against vegetative forms of anthrax bacteria and do not cause the death of anthrax spores. In addition, in order to suppress the vital activity and death of vegetative cells of anthrax or bacteria, it takes a lot of time and optimal conditions for the life of actinomycetes and favorable conditions for germination of anthrax spores: the presence of a nutrient substrate in the soil, soil temperature not lower than 18-20 ° С, soil moisture 80 -90% and others.

There is also a method of disinfecting the soil (copyright certificate No. 376074, IPC A01N 7/04), which consists in stretching a film over the infected area parallel to the surface of the earth, the edges of which are dug up with earth, and open vessels are installed under the film into which liquefied gas is poured, for example a gas mixture of ethylene oxide with methyl bromide (OKEMBM). In order to increase the effectiveness of disinfection, open pits are installed in the soil.

The main disadvantages of the method are:

- low efficiency of the disinfection method, because the process itself depends on external conditions (temperature, humidity and soil structure);

- high risk of emergency situations at any stage of the work on soil disinfection in the form of leakage or release of toxic gases into the atmosphere;

- air and soil pollution with toxic gases upon completion of the disinfection process with the remnants of the gas mixture, etc.

The closest technical solution to the claimed invention is a method of disinfecting the territories of old anthrax cattle cemeteries, (Pryanichkova V.V., Bikbulatov I.Kh., Bakhonina E.I. Disinfection of the territory of old cattle cemeteries // Journal of Life Safety. - 2013, - № 11., - pp. 16-20), in which the soil is watered with 0.7% (hydrogen peroxide) with an aqueous solution of “SaBiDEZ” of the following composition. %:

- medical hydrogen peroxide - 3.3-3.8;

- Alkyldimethylbenzylammonium chloride (ADBAH) - 1.4-1.6

- water - the rest

The main disadvantages of the method are:

- the absence of a disinfecting effect on the surface of the soil during decontamination of the soil from anthrax spores;

- low efficiency of the disinfection method in the disinfection of soil cotaminated with bacteria of pathogens of dangerous and especially dangerous infections (plague, cholera, tularemia, etc.), on the surface and to a shallow depth of soil;

- long-term processing (watering) of the soil with solutions of the product leads to the accumulation of chemical components of the product in the soil, etc.

In this regard, an important and urgent task of medicine and veterinary medicine is the development of a highly effective disinfectant and disinfection method designed for the accelerated and reliable elimination of soil foci of anthrax resulting from the burial, slaughter, and mortality of sick animals as a result of technological accidents and catastrophes or bioterrorist acts .

The technical result of the invention is to increase the efficiency of disinfection of soil contaminated by bacterial spores of the causative agent of anthrax or bacteria of causative agents of dangerous and especially dangerous infections, with short periods of exposure; ensuring the safety of soil disinfection, eliminating the accumulation of hazardous substances, as well as their decomposition products in the soil or in the environment, reducing the dependence of the effectiveness of the soil disinfection process on ambient temperature, humidity and soil structure; the presence of high stability of the disinfectant and its working solutions.

The technical result is achieved due to the fact that a plot of soil contaminated with anthrax pathogen spores or bacteria of pathogens of dangerous and especially dangerous infections is separated from uncontaminated soil with an artificial trench 500-600 mm wide and 300-400 mm deep to prevent the disinfectant solution from draining out of the plot , and the soil surface of the plot is dug up (loosened) or holes are drilled in the soil to a depth of 500 mm at the rate of 4-6 pits per 1 m ² , then the soil surface is watered once or twice 10-15% (by hydrogen peroxide) with a disinfectant solution, including medical hydrogen peroxide, alkyldimethylbenzylammonium chloride (ADBAH); calcium chloride and sulfanol in the following components, wt. %:

Medical hydrogen peroxide - 30.0 ± 3.0;

Alkyldimethylbenzylammonium chloride - 2.0 ± 0.2;

Calcium chloride - 1.0 ± 0.1;

Sulfanol - 1.0 ± 0.1,

Water - the rest at the rate of 10-15 liters per 1 m with an interval between treatments of 24 hours with a total exposure time of 24-72 hours.

The proposed disinfectant and method of disinfecting soil contaminated by anthrax spores or bacteria of causative agents of dangerous and especially dangerous infections, in contrast to the prototype, has a wide spectrum of antimicrobial activity, sufficient disinfecting efficiency for decontamination of the soil from bacteria and bacterial spores, effective soil disinfection can be achieved with a wide range of ambient and soil temperatures (from 4 to 40 ° C). The proposed tool and method allows you to inactivate bacteria and bacterial spores to a depth of 500 mm of soil at low cost (24-72 hours).

The components of the proposed disinfectant and the products of its decay, in contrast to the prototype, after one or two tillages after 5-10 days are absent in the soil.

The proposed tool and method for disinfecting the soil, provide effective, environmentally friendly inactivation of spores and bacteria of pathogens of dangerous and especially dangerous infections.

Example 1. To determine the antimicrobial and disinfecting effect of the means and method of disinfecting the soil contaminated with anthrax spores, use a disinfectant of the following composition, mass. %:

Medical hydrogen peroxide - 30.0;

Alkyldimethylbenzylammonium chloride - 2.0;

Calcium chloride -1.0;

Sulfanol 1.0

Water - the rest

An area of loamy soil with an area of 10 m ² contaminated with spores of the vaccine strain of bacteria of the causative agent of anthrax Bacillus anthracis piece ST-1 to a depth of not more than 50 mm at the rate of 2.0 × 10 ³ CFU / cm is delimited from the uncontaminated soil with a trench 600 mm wide and 400 mm deep to prevent the disinfectant from draining outside the infected area. The soil is disinfected by watering the surface of the soil with a 15% solution of the agent at the rate of 15 l per 1 m ² at an exposure time of 24 hours. The soil temperature was 8 ° C.

As a control, we used soil samples contaminated with spores of a vaccine strain of bacteria of the causative agent of anthrax Bacillus anthracis pcs. TI-1 at a concentration of 2.0 × 10 ³ CFU / cm. The processing of soil samples in control experiments was carried out with sterile water at the rate of 15 l per 1 m ² .

To assess the disinfecting effect of the disinfectant, the soil sample was neutralized with a universal neutralizer, then bacterial spores were isolated from the soil and the fish meal hydrolyzate (GRM) was sown in solid medium and thermostated at 37 ° C for 7 days. Microbiological studies of soil samples treated with a disinfectant showed that living anthrax spores are absent on the soil surface and at a soil depth of 50 mm (table 1). Microbiological studies of soil contaminated with anthrax bacteria spores in the control spilled with sterile water showed the presence of live anthrax bacteria spores in soil samples in an amount of approximately 2.0 × 10 ^{2 = 3} CFU / cm (Table 1).

The presented results in table 1 show that the proposed tool and method of soil disinfection allows for a one-time filling of the soil with 15% (hydrogen peroxide) disinfectant solution at the rate of 15 l / m ² at an exposure of 24 hours to inactivate spores of anthrax bacteria to a soil depth of up to 50 mm .

Example 2. To determine the sporocidal and disinfecting effect of the means and method of disinfecting the soil, use a disinfectant of the following composition, mass. %:

Medical hydrogen peroxide - 27.0;

Alkyldimethylbenzylammonium chloride - 1.8;

Calcium chloride - 0.9;

Sulfanol - 0.9

Water - the rest

A plot of chernozemic soil with an area of 10 m ² contaminated by bacterial spores of the causative agent of anthrax to a soil depth of not more than 500 mm is separated from uninfected soil with a trench 500 mm wide and 400 mm deep, and the soil surface of the plot is dug up (loosened) by one bayonet shovel without turning over the soil , decontaminated by double irrigating the soil surface 13% (hydrogen peroxide) solution means the rate of 150 l / 1 m ² (300 l / 1 m ³⁾ with an interval between treatments 24 hours at a total exposure time of 72 hours. The soil temperature during and tests carried was 4 ° C.

To assess the disinfecting effect, a soil sample was neutralized with a universal neutralizer, then a sample of bacterial spores from the soil was sown on a solid timing medium and thermostated at 37 ° C for 7 days. To control the soil, chernozem contaminated with anthrax spores was also poured with sterile water at the rate of 150 l / 1 m ² (300 l / 1 m ³ ), and the isolated spores from the soil were sown on solid timing medium and thermostated at 37 ° C for 7 days.

Microbiological research methods found that in soil samples contaminated with bacterial anthrax spores taken on the soil surface and at a depth of 500 mm, after two-time treatment with a disinfectant solution, live spores are completely inactivated (table 2). In control soil samples after pouring with sterile water, anthrax spores in an amount of approximately 2.0x10 "CFU / cm ^{3 were} determined viable (table 2).

So, it was found that the proposed peroxide composite disinfectant and method of soil disinfection inactivate anthrax spores on the soil surface and to a soil depth of up to 500 mm.

Example 3. To determine the sporocidal and disinfecting effect of the agent and method for disinfecting soil, use a disinfectant of the following composition, mass. %:

Medical hydrogen peroxide - 33.0;

Alkyldimethylbenzylammonium chloride - 2.2;

Calcium chloride -1.1;

Sulfanol 1.1

Water - the rest

A plot of podzolic soil with an area of 10 m ² contaminated with bacterial spores of the causative agent of anthrax to a depth of not more than 300 mm is delimited from uninfected soil with a trench 600 mm wide and 400 mm deep, and they are drilled into the soil with a mechanical drill to a depth of no more than 500 mm by 4 -6 pits per 1 m, disinfected by watering the surface of the soil with a 10% solution of the agent (using hydrogen peroxide) at the rate of 100 l per 1 m (300 l / 1 m) at an exposure time of 24 hours. The soil temperature was 8 ° C.

To assess the disinfecting effect, a soil sample was neutralized from a disinfectant with a universal neutralizer, then a bacterial spore sample was sown from the soil on a solid timing medium and thermostated at 37 ° C for 7 days.

Samples of podzolic soil were used as a control,

contaminated with anthrax spores spilled with sterile water instead of a disinfectant. Spores from control soil samples were plated on a tight timing medium and thermostated at 37 ° C for 7 days.

Microbiological studies of the disinfecting effect of the proposed disinfectant and method of disinfecting soil areas showed that when soil is poured with 10% peroxide disinfectant solution until the soil is completely wetted to a depth of 300 mm, anthrax spores are completely inactivated (table 3). In control soil samples, on the contrary, the viability of anthrax spores is maintained at the level of initial soil contamination by spores.

Thus, it was found that with a one-time pouring of soil contaminated with anthrax spores with a 10.0% solution of the peroxide composition during 24 hours exposure, bacterial spores are inactivated in the soil to a depth of 300 mm.

Example 4. To determine the sporocidal and disinfecting effect of the means and method of disinfecting the soil, use a disinfectant of the following composition, mass. %:

Medical hydrogen peroxide - 30.0;

Alkyldimethylbenzylammonium chloride - 2.0;

Calcium chloride -1.0;

Sulfanol 1.0

Water - the rest

A section of clay soil with an area of 10.0 m ² contaminated with bacterial spores of Bacillus stearothermophilus ATCC 12980 to a depth of 300 mm, simulating bacterial spores of the causative agent of anthrax, is separated from uninfected soil with a trench 500 mm wide and 400 mm deep, and the soil is drilled using a mechanical drill a depth of not more than 300 mm, 4-6 pits per 1 m, disinfected by watering the surface of the soil with 11% solution of the product at the rate of 100 l per 1 m ² (300 l / 1 m ³ ), and exposure time 24 hours. The soil temperature was 3 ° C.

To assess the disinfecting effect, soil samples were neutralized with a universal neutralizer, then samples of B.stearothermophilus ATCC 12980 bacterial spores from the soil were sown on a solid Hottinger medium and thermostated at 37 ° C for 7 days. Microbiological studies have shown that in the soil treated with an 11.0% disinfectant solution, there are no live spores of B. stearothermophilus ATCC 12980 to a depth of 300 mm.

Example 5. To determine the bactericidal and disinfecting effect of the agent and method for disinfecting soil, use a disinfectant of the following composition, mass. %:

Medical hydrogen peroxide - 30.0;

Alkyldimethylbenzylammonium chloride - 2.0;

Calcium chloride -1.0;

Sulfanol 1.0

Water - the rest

Clay soil area of 10.0 m contaminated with bacteria Escherichia coli pcs. K-12 to a depth of 200 mm, imitating the bacteria of causative agents of dangerous and especially dangerous infections, is separated from non-contaminated soil with a trench 500 mm wide and 400 mm deep, and the soil is drilled using a mechanical drill to a depth of not more than 300 mm, 4-6 pits per 1 m, they are disinfected by watering the surface of the soil with a 10% solution of the agent at the rate of 100 l per 1 m ² (300 l / 1 m ³ ), and the exposure time is 24 hours. The soil temperature was 5 ° C.

To assess the disinfecting effect, soil samples were neutralized with a universal neutralizer, then bacteria samples of E. coli pcs. K-12 from the soil was sown on a dense medium MPA and thermostated at 37 ° C for 7 days. Microbiological studies have shown that in the soil treated with a 10.0% disinfectant solution, living bacteria E. coli pc are absent to a depth of 200 mm. K-12 spores.

Thus, laboratory and experimental field tests of the means and method of disinfecting soil areas showed that the proposed tool and method of disinfecting the soil provide complete decontamination of the soil to a depth of 500 mm from spores of bacteria of the causative agent of anthrax and from bacteria of causative agents of dangerous and especially dangerous infections. The tool and method for disinfecting soil are environmentally friendly.

The use of a means and method of disinfecting areas of soil contaminated with anthrax pathogen spores or bacteria of pathogens of dangerous and especially dangerous infections resulting from the burial, slaughter, and mortality of sick animals as a result of industrial accidents and catastrophes or bioterrorist acts will allow for the first time the sanitary-epidemiological and sanitary - Veterinary services to effectively and safely carry out sanitary and epidemiological measures to eliminate soil foci of dangerous and especially dangerous facies, including soil foci of anthrax.

Claims (7)

1. A means of disinfecting areas of soil contaminated with bacterial spores of the causative agent of anthrax or bacteria of causative agents of dangerous and especially dangerous infections, including medical hydrogen peroxide, alkyl dimethylbenzylammonium chloride (ADBAH) and water, characterized in that it additionally contains calcium chloride and sulfanol in the following components mass. %: Medical hydrogen peroxide - 30.0 ± 3.0; Alkyldimethylbenzylammonium chloride - 2.0 ± 0.2; Calcium chloride - 1.0 ± 0.1; Sulfanol - 1.0 ± 0.1, Water is the rest. 2. The method of disinfecting areas of soil contaminated with bacterial spores of the causative agent of anthrax or bacteria of causative agents of dangerous and especially dangerous infections, to a depth of 50 to 500 mm, with a tool according to claim 1, including watering the area of infected soil, characterized in that the area of infected soil is limited from uncontaminated soil with a trench 500-600 mm wide and 400 mm deep, the soil surface of the site is dug up to a depth of 200 mm or pits are drilled into the soil to a depth of 500 mm at the rate of 4-6 pits per m ² and disinfected by a single or double watering the surface of the soil with 10-15% solution of a disinfectant (hydrogen peroxide) at the rate of 15-150 liters per 1 m ² , with an interval between treatments of 24 hours with a total exposure time of 24-72 hours and at ambient and soil temperatures of 4 to 40 ° C.