Method for bioorganic purification of water bodies using bacteria and specific bacteriophages

Abstract

FIELD: biotechnology.SUBSTANCE: method for bioorganic purification of water bodies using bacteria and specific bacteriophages is proposed, including preliminary activation of the biological preparation “Biofermentil” based on the bacteria Bacillussubtilis and Bacilluslicheniformis in water with sugar at the rate of 150 g of the preparation per 450 g of sugar and per 10 liters of water per 1 hectare of water surface for 48 hours, and when the water in the reservoir warms up above 14 degrees Celsius, the activated biological preparation is added to the reservoir; after 14 days, the biological preparation “Biofermentil” is prepared again and added; 7–10 days after repeated application of the biological preparation “Biofermentil”, a bacteriophage with virulence from 70 to 95% against Pseudomonadaceae is added at a concentration of 108 to 1010 at the rate of 1 liter of the preparation per 1 hectare of water surface, evenly distributed over the surface of the reservoir.EFFECT: improving the ecological state of a reservoir by reducing silt deposits and pathogens that cause fish diseases.1 cl, 1 ex

Description

[0001] The invention relates to technologies for cleaning water bodies, as well as to industrial fish farming and keeping aquatic organisms in installations with a closed water supply system and adjustable environmental parameters.

[0002] Water quality is a pressing issue. Currently, the most common methods used are chemical treatments—lime, chlorine, and disinfectants. Mechanical methods, such as excavator and dredger cleaning, are also used.

[0003] A method of water treatment for cultivating aquatic organisms in closed volumes and a device implementing it are known (Patent 2721534, published: 2020.05.19), characterized in that a mixture of probiotic microorganisms Bacillus subtilis is added to the pool, the water is aerated with ascending flows of the water-air mixture from the bottom of the pool, when the total ammonium nitrogen content in the water reaches zero, aquatic organisms are planted in the pool and their cultivation begins, during the cultivation process, the level of total ammonium nitrogen, dissolved oxygen and the amount of activated sludge are monitored, when the threshold values ​​​​of activated sludge are reached, water with particles of activated sludge begins to be drained from the pool, to replenish the drained water, clean water is added to the pool until the amount of activated sludge is brought to safe values ​​​​for the cultivated aquatic organisms,In this case, carbon-containing substances are periodically added to the water of the pool to feed the Bacillus subtilis microorganisms, in a ratio of 6-20 g of carbon per 1 g of nitrogen in the water until the level of total ammonium nitrogen is reduced to a safe value for the grown aquatic organisms; when the maximum density of aquatic organisms is reached, they are transferred to other pools.

[0004] The disadvantage of this method is its limited effectiveness.

[0005] The technical problem that the present invention is aimed at solving is the expansion of the existing arsenal of known effective methods for cleaning water bodies.

[0006] The technical result consists of improving the ecological state of the reservoir, processing soft silt and organic matter, reducing pathogens (through bacteriophages) that cause diseases, and inhibiting their growth through bacteria. It is this combination that allows for the most favorable results.

[0007] The stated problem is solved by the fact that in the declared method of bioorganic purification of various water bodies using bacteria and specific bacteriophages, according to the declared technical solution, bioorganic purification of various water bodies is carried out using bacteria and specific bacteriophages, while the compositions of the preparations are selected based on a preliminary study of the reservoir condition, samples of sludge, pathological material, water, algae; when the water of the reservoir warms up above 14 degrees Celsius, a biological preparation is prepared, for which a preparation based on Bacillus subtilis and Bacillus licheniformis bacteria is preliminarily soaked in water with sugar at the rate of 150 grams of the preparation per 450 grams of sugar per 10 liters of water per 1 hectare of water surface for 48 hours and introduced into the reservoir;After 14 days, when the positive microflora occupies the maximum amount of space in the reservoir and begins to prevail, an identical repeated preparation and introduction of the biological preparation is carried out; 7-10 days after the repeated introduction, when the growth of the introduced bacteria calms down, a bacteriophage with virulence from 70 to 95% in relation to Pseudomonadaceae and Aeromonas hydrophila is introduced at a concentration of 10; 8 up to 10 10 at a rate of 1 liter of the product per hectare for uniform distribution over the surface of the reservoir; the combination of a biological product based on the bacteria Bacillus subtilis and Bacillus licheniformis with the use of highly virulent bacteriophages ensures a prolonged effect on the reservoir's health.

[0008] The claimed method of bioorganic purification of various water bodies using bacteria and specific bacteriophages consists in the fact that bioorganic purification of various water bodies is carried out using bacteria and specific bacteriophages, while the compositions of the preparations are selected individually depending on the results of the conducted research into the problems of the water body.

[0009] They conduct a study of the condition of the reservoir, for this they take samples of silt, pathological material, water, algae and carry out bacterial culture on a nutrient medium in order to identify pathogens and determine the total microbial count.

[0010] When the reservoir water warms above 14 degrees Celsius, a biological product is prepared. A product based on the bacteria Bacillus subtilis and Bacillus licheniformis is first soaked in water with sugar at a rate of 150 grams of product per 450 grams of sugar per 10 liters of water per hectare of water surface for 48 hours and then added to the reservoir. The concentration of the bacteria in the dry form has a titer of 10 9 When started on water containing sugars, the product is activated and emerges from the spore state. Then, after 14 days, when the positive microflora has occupied the maximum amount of the pond's space and begins to predominate, an identical second preparation and application of the biological product are performed.

[0011] The ratio of Bacillus subtilis and Bacillus licheniformis bacteria is selected based on the depth of the pond and the various functions of the bacteria, which enable them to combat pathogenic bacteria more effectively. For example, in ponds with depths greater than 3.5 meters, Bacillus licheniformis is primarily selected (at least 65% of the total mix), as it requires very little oxygen to survive. However, in ponds with depths less than 3.5 meters, Bacillus subtilis is used in the same quantity, as it is more active in high oxygen levels. It is crucial that the different bacterial strains in the preparation do not conflict with each other, but rather enhance each other's effects.

[0012] When released into a body of water, bacteria digest organic matter in the water—plant remains, algae, feces, and various nitrogenous compounds. This process breaks down the nutrient medium that serves as food for pathogenic flora, and also halts oxidative and putrefactive processes that cause fish disease and death.

[0013] Seven to ten days after repeated application, when the growth of the introduced bacteria has subsided (they have become spore-forming and their oxygen consumption from the water has significantly decreased), a specific bacteriophage with 70 to 95% virulence against Pseudomonadaceae and Aeromonas hydrophila is added. The bacteriophage is selected based on the problem identified in laboratory studies. Laboratory strains of bacteriophage with the required activity: strain P-3087 / 2-2, strain P-3252 / 8 / 3-2, strain P-3252 / 6 / 4-1, strain P-3071 / 1-2, have been isolated and stored in the laboratory of poultry diseases of the Siberian Federal Scientific Center of the Russian Academy of Sciences.

[0014] To suppress Pseudomonadaceae (lat.) - a family of bacteria from the class Gammaproteobacteria, including the genera Azomonas, Azomonotrichon, Azorhizophilus, Azotobacter, Cellvibrio, Mesophilobacter, Pseudomonastypus, Rhizobacter, Rugamonas and Serpens and others, found in a reservoir. If their concentration in reservoirs significantly exceeds 10 4 and 10 7 select a bacteriophage (strain P-3087 / 2-2, strain P-3252 / 8 / 3-2, strain P-3252 / 6 / 4-1, strain P-3071 / 1-2) with a virulence of at least 80% at a concentration of 10 8 up to 10 10 In such concentrations, 1 liter of the preparation, evenly distributed over the surface of the reservoir, is sufficient per hectare of water.

[0015] In the case of Aeromonas hydrophila (Latin) detected in the reservoir, a heterotrophic gram-negative rod-shaped bacterium that lives primarily in areas with warm climates. It can be found in both fresh and salt water. A bacteriophage virulence of 70% and a concentration of 10 are sufficient. 9 .

[0016] In this combination, Bacillus subtilis and / or Bacillus licheniformis bacteria inhibit the growth of pathogenic flora, and bacteriophages reduce the number of a specific pathogenic strain.

[0017] This combination of a biological product with highly virulent bacteriophages achieves a prolonged, healthy effect on the reservoir for at least one year. If pathogenic strains are not introduced into the reservoir from outside, the effect can last for several years. As a result, the reservoir's condition and productivity are improved. The risk of fish disease and mortality is reduced. Economic indicators improve by 20 to 60%. The result is an environmentally friendly product with a low production cost on an industrial scale and excellent taste.

[0018] Practical implementation example.

[0019] The studied water body was analyzed for pathogenic factors. Contamination by Pseudomonadaceae and Aeromonas hydrophila was detected. In the spring, when the pond water warmed to 14 degrees Celsius, the preparation "Biofermentil" consisting of Bacillus subtilis (hay bacillus) and Bacillus licheniformis bacteria strains was added after being cultured in water containing sugars.

[0020] The ratio of bacteria is 2 strains of Bacillus subtilis (hay bacillus) and 1 strain of Bacillus licheniformis in equal parts (33%) and a concentration of 10 9(Bacteria should interact and enhance their properties, not suppress them) was selected based on the average depth of the reservoir, 1.5 meters. The solution was prepared at a ratio of 1 part Biofermentil to 3 parts sugar. The dosage for 1 hectare of water surface was calculated as 150 grams of the product per 450 grams of sugar per 10 liters of water. This solution preparation releases the bacteria from the spore state and increases the titer by one degree. The product is ready for use after 48 hours.

[0021] The liquid preparation is distributed more evenly throughout the reservoir compared to the dry one.

[0022] After 14 days, the procedure was repeated to maximize the displacement of pathogenic flora and the processing of organic matter that had accumulated over the winter.

[0023] After using Biofermentil, bacteriophage strain P-3087 / 2-2 was added. Adding bacteriophages without Biofermentil is unacceptable, as in the absence of competing microflora, Pseudomonas strains resistant to these bacteriophages will grow, leading to a renewed outbreak. These processes do not occur with the addition of competing beneficial microflora.

[0024] The bacteriophage used should be at a concentration of 10 8 up to 10 10 Depending on the number of pathogenic strains. At these concentrations, the dosage is 1 liter of the product per hectare of water surface.

[0025] In mid-August, Biofermentil is applied again to increase the bacterial concentration in the water for the fall at a dosage of 150 grams per hectare of water surface. This is because algae will die off in the fall and need to be processed before winter to prevent rotting (prevention of algae freezes).

[0026] In severe situations or in countries with year-round hot climates, the drug "Biofermentil" is applied once a month.

[0027] In total, in the fall we observe a decrease in Pseudomonas titers from 10 6 up to 10 3 or lower. This is enough to stop fish diseases and deaths.

[0028] The reservoir has seen a reduction in silt deposits (compression of soft silts), an increase in oxygen, and an almost complete reduction in fish diseases.

[0029] Increases reservoir productivity from 20 to 70%.

[0030] Reduces financial and labor costs in fish farming.

[0031] Improves the organoleptic taste of fish. The result is an environmentally friendly product.