RU2204597C2 - Strain of archaebacterium bacillus species vkpm b-5061 as decomposing agent petroleum products and phosphate used in soil and sewage treatment - Google Patents

Abstract

FIELD: industrial microbiology, ecology. SUBSTANCE: invention relates to microbiological method of treatment of sewage and soil from petroleum products and phosphates. The strain Bacillus sp. VKPM B-5061 is able to purify sewage from petroleum products and phosphates under conditions of low and high temperatures (from +15 C to +90 C) normalizing their content in flows to standardized norms of permissible concentrations both in volley and standard regime of treatment of sewage before their feeding to refinery units and to reservoir finally and to soil up to background concentration of petroleum products in 21 days after their application. Culture Bacillus sp. VKPM B-5061 shows no demanding upon to nutrient media and can be cultured under both aerobic and anaerobic conditions. To enhance effectiveness of the strain Bacillus sp. VKPM B-5061 productivity the strain is sorbed on woven carrier (kapron fabric), installed in sewage flow before refinery equipments and bond its in soil with spent silt by applying on surface that allows to retain high efficiency of working ability of microorganisms in flow-type system of chemically polluted sewer system under chemostatic conditions. EFFECT: enhanced effectiveness of strain.

Description

 The strain is deposited in the All-Union Collection of Industrial Microorganisms (VKPM) of the Institute "Research Institute of Genetics" of the Russian Ministry of Health.

 The invention relates to biotechnology, in particular to microbiological treatment of soil and wastewater, the use of a strain of a microorganism for the destruction of petroleum products and phosphates in wastewater and soil and can be used to clean the soil and wastewater of petrochemical enterprises.

 A known bacterial strain is a destructor of the biogens of nitrogenous and carbon compounds in the wastewater of chemical plants (the bacterial strain Pseudomonas sp. N-114 is an ammonia and urea destructor). Auth. testimonial. SU 1735210 Al.

 The disadvantage of this strain is the impossibility of its use at high temperatures for the degradation of phosphorus biogens.

Known strains of prokaryotic organisms of non-photosynthetic bacteria of spore-forming species (Bacillus acidocaldarius, Bacillus sp.) - author Loginova L.G. // Biology of thermophilic microorganisms. M .: Nauka, 1986, p.5, v. 22, used for the degradation of petroleum products (normal alkanes). The disadvantage of these strains is the inability to use them for wastewater treatment in extreme conditions of synthetic rubber plants, since the runoff of these industries has a high temperature reaching 75-95 ^o C.

 The closest in technical essence to the claimed strain is a bacterial strain Bacillus stearothermophilus, isolated from active sludge of wastewater from petrochemical industry enterprises, used for microbiological treatment of wastewater from normal alkanes.

 The disadvantage of the proposed strain is its inability to work at high temperatures, as well as with a joint high concentration of normal alkanes with a high concentration of phosphates, because it causes gelation in the systems, leading to plugging of process pipelines.

 The aim of the invention is to obtain a strain - an effective destructor of oil products and phosphates, in the corresponding wastewater of petrochemical enterprises, by using microorganisms from the genus Bacillus adsorbed on tissue carriers - non-photosynthetic bacteria of prokaryotic organisms - thermophiles adapted to high temperature conditions and selectively degrading normal alkanes of synthetic production rubber.

 To achieve this goal, a strain of archaebacteria Bacillus sp. VKPM V-5061, deposited in the All-Union Collection of Industrial Microorganisms (VKPM) of the Institute "Research Institute of Genetics" of the Ministry of Health and Industry of the Russian Federation and intended as a destructor of oil products and phosphates.

 The strain is placed on woven carriers (nylon sheets) installed in the wastewater stream in front of the treatment plant.

 The proposed fabric allows you to maintain high efficiency microorganisms in the flow system of chemically contaminated sewers in a chemostat.

Exposure of the tissue carrier at low temperatures (from -15 ^° C to 0 ^° C) did not adversely affect the growth of the test culture.

The resulting strain of Bacillus sp. VKPM B-5061 is characterized by the following morphological, cultural and physico-biochemical characteristics:
CULTURAL-MORPHOLOGICAL SIGNS:
Bacterial prokaryotic cells - rounded gram-negative polymorphic rods, motile (moving with the help of polar flagella), form spores and capsules. The cell size is 0.6 • 1.0 • 1.2. Inclined to settle down singly. Cells are inactive, have a limited number of flagella.

 Culture of Bacillus sp. VKPM B-5061 is undemanding to nutrient media and can be cultivated under aerobic conditions on ordinary plate and beveled meat-peptone agar (meat-peptone broth), and under optional anaerobic conditions when sown in a short column of meat-peptar agar under a layer of petroleum jelly (to prevent vaseline oil) (to prevent .

Colonies of this strain on agar (90 ^° C; 24 hours) are round with a diameter of up to 1.5 mm, convex, moist, not tending to merge, slightly phosphorizing, but not staining the medium.

In the meat and peptone broth (90 ^o C; 24 h), uniform turbidity is observed without film formation with the gradual formation of a slight, easily swollen sediment with a golden yellow tint.

PHYSIOLOGICAL-BIOCHEMICAL CHARACTERISTICS:
Anaerobic, with a high growth temperature (up to 90 ^o C), the optimum growth temperature is 30-75 ^o C, pH 6.2-8.6. At neutral pH, significant starch thinning was observed. It has catalase, oxidase and urease activity. It decomposes oil products and phosphates.

Short-term storage of the strain is carried out on mowed agar in a refrigerator with weekly reseeding. The culture after storage is refreshed by reseeding and germinated in a thermostat at 85 ^o C during the day until a distinct plaque on the potato plates.

 For mass propagation of the culture, the method of flowing chemostat was used. The limiting intracellular concentration of phosphorus content varied in the following ranges (from 4.2–8.7 mg / L to 13.7–16.2 mg / L).

 At phosphate concentrations up to 15 mg / l, the biomass content exceeds 10 mg / l, while latex particles are observed in the biomass. The decrease in phosphorus in the medium has a direct effect on the synthesis of total RNA, since it is a structural component of RNA. The lack of phosphorus in the environment contributes to the accumulation of polysaccharides in cells, which significantly increases the oxidizing ability of cells.

 The effect of destruction is tested on a number of effluents of chemical plants. The properties of the strain to degrade petroleum products and phosphates are illustrated by examples.

Example 1. To one liter of wastewater containing 1370 mg / l of petroleum products and 16.2 mg / l of phosphates, 10 ml of a suspension of the test microorganism strain washed with 5 ml of tap water from potato agar cubes was added. The sample contained 70 million microbial cells in 1 ml. After incubation of the strain in a thermostat at 90 ^o C for 24 hours, the remainder of the oil products amounted to 739.8 mg / l or 54% of the initial level. The destruction of phosphates took place from an initial concentration of 16.2 mg / L to a final concentration of 7.6 mg / L, which is 47%.

Example 2. To one liter of water containing 153 mg / l of petroleum products and 8.7 mg / l of phosphates, suspensions of the tested microorganism strain were added. The sample contained 25 million microbial cells in 1 ml. After the strain was incubated in a thermostat at 90 ^° C for 24 hours, the residue of petroleum products amounted to 41.3 mg / L or 27%, and the final concentration of phosphates was 2.7 mg / L, which corresponds to 31% of the initial level.

 Example 3. In the actual production conditions of the Togliatti AO Sintezkauchuk, having an initial concentration of oil products in the stock of 12.72 mg / l, they are uniformly degraded before entering treatment plants in sewage systems using a strain-destructor and shows the final concentration of oil products 9, 67 mg / l. At the same time, the concentration of microbial suspension in the wastewater sample was 1.5 billion microbial bodies per 1 ml, and the concentration of phosphates was: at the inlet 12.63 mg / l, and at the outlet 4.67 mg / l, which is expressed as a percentage 24 and 37% respectively.

 Example 4. In the flow system of workshop E - 2 of Sintezkauchuk JSC, a tissue carrier with a culture of the microorganism Bacillus sp. VKPM B-5061 for a period of more than three months, while the number of microorganisms in the smear from the tissue was 230 thousand microbial bodies in 1 ml of waste water. As for pathogenicity, the proposed strain has less pathogenicity compared to existing ones and with E. coli.

 Toxicological tests of the pathogenicity of the proposed strain were carried out on a culture of white mice with a standard for comparison of E. coli, easily tolerated by intraperitoneal infection.

 From these examples it follows that the applied strain of Bacillus sp. VKPM B-5061 has a higher, 2–3 times degrading activity, in relation to phosphates and oil products. The advantages of the strain are a high degree of destruction of oil products and phosphates both in salvo and in the normal mode of wastewater treatment before supplying them to treatment plants and, ultimately, to a reservoir.

Claims (1)

 The strain of archaebacteria Bacillus sp. VKPM V-5061 is a destructor of oil products and phosphates used in the treatment of soil and wastewater.