RU2269571C1 - Bacterium desulfovibrio sp. strain useful in wastewater purification from heavy metal ions - Google Patents

Abstract

FIELD: industrial microbiology, in particular new sulfide-producing bacterial strain.

SUBSTANCE: application of bacterium Desulfovibrio sp. strain for wastewater purification from heavy metal ions is disclosed. Claimed strain is isolated from heavy metal ion-containing wastewater of galvanic industrial enterprises. Purification method has effectiveness of about 99.9 %.

EFFECT: method for wastewater purification from heavy metal ions of improved effectiveness.

Description

The invention relates to industrial microbiology and relates to the production of a new strain of bacteria producing sulfide, which can be used in biotechnology for the treatment of industrial wastewater from engineering, instrument-making, and electrical enterprises.

It is known about the use of sulfate reducing bacteria (SBB) for wastewater treatment from heavy metal ions. In the process of biochemical purification, heavy metal ions are bound by biogenic hydrogen sulfide to insoluble sulfides, as well as the reduction of certain metals and precipitation in the form of hydroxides. To increase the efficiency of biochemical purification methods using SVB, a high level of bacterial growth activity is required.

To create the necessary concentration of microorganisms in the process of wastewater treatment from heavy metal ions, various methods are used, or easily digestible organic food is added for SVB [copyright certificate No. 857013, cl. Since 02 F 3/28, 1979], or use specially selected biocenoses of microorganisms [copyright certificate No. 941319, class. C 02 F 3/34, 1980].

However, the microorganisms used do not provide special activity for the growth of SFS in industrial wastewater, characterized by a different degree of pollution and a different composition of polluting components.

The prototype was a bacterial strain Desulfovibrio baarsii EGAST-6, used for wastewater treatment from heavy metal ions. This strain is a highly active strain of sulfate-reducing bacteria, providing a stable increase in biomass during wastewater treatment from heavy metal ions [RF patent No. 2150502, class. C 02 F 3/34, C 12 M 1/20, bull. No. 16, 2000].

The strain Desulfovibrio baarsii EGAST-6 isolated from industrial runoff containing cutting fluids.

The strain Desulfovibrio baarsii EGAST-6 was deposited in the All-Russian collection of microorganisms (VKM) of the Institute of Biochemistry and Physiology of Microorganisms under the number VKM B-2025 D.

The strain EGAST-6 Desulfovibrio baarsii was identified: Bergey's Manual of Systematic Bacteriology "; Edit .: Krieg N.R., Holt J.G. - Baltimore, Williams Wilkins, 1984, V.1, 964 p.

This strain is an active producer of hydrogen sulfide for the precipitation of heavy metal ions from solutions and is characterized by the following cultural and morphological characteristics.

Cultural and morphological characteristics of the strain are determined during its cultivation on standard media, the recipes of which are taken from the catalog of the All-Russian collection of microorganisms No. 92 and No. 109 and are given below in the publication "All-Russian collection of microorganisms. Catalog ”, Pushchino-Moscow, VNITI, 1992, 362 p. Cultivation is carried out under anaerobic conditions at 30 ° C for two days.

Cultivation conditions:

- cultivation is carried out under anaerobic conditions at 30 ° C for two days;

- the optimal pH of 7.2 ... 7.4;

- used environment: 63 DSM.

Morphological features and cell cytology:

- under microscopy, the cells are represented by single vibrios with a size of 0.5 ... 1.0 and 1.5 ... 2.0 microns;

- Gram stain - negative;

- they do not form a dispute.

The physiology properties of the strain:

- strict anaerobic;

- acetate, propionate, butyrate are used as hydrogen donors and carbon sources.

Biochemical characteristics of the strain:

- cells contain cytochrome C.

Storage conditions:

- preserved by reseeding on the cultivation medium;

- can be stored in the refrigerator at 4 ... 10 ° C for 3 ... 6 months.

Prescriptions of media from the catalog of the All-Russian Collection of Microorganisms: N 92. Medium with lactate for Desulfovibrio.

Solution 1:

K ₂ HPO ₄ 0.5 g NH ₄ Cl 1.0 g CaCl ₂ · 6H ₂ O 0.1 g MgSO ₄ · 7H ₂ O 2.0 g Na ₂ SO ₄ 1.0 g sodium lactate 5.0 g yeast extract 1.0 g resazurin 0.001 g cysteine 0.5 g distilled water 950.0 ml

Solution 2:

NaHCO ₃ 4.0 g distilled water 40.0 ml

Solution 3:

Na ₂ S · 9H ₂ O 0.3 g distilled water 6.0 ml

Solution 4:

FeSO ₄ · 7H ₂ O 0.4 g distilled water 10.0 ml

Preparation: bring solution 1 to a boil while blowing through a solution of an oxygen-free gas mixture consisting of 97% N ₂ and 3% H ₂ . Sterilize these gases in the atmosphere. Solution 3 is sterilized in an atmosphere of N ₂ . After sterilization, connect the solutions. The pH of the medium after mixing is set to 6.8.

N109. Wednesday

Solution 1:

K ₂ HPO ₄ 0.5 g NH ₄ Cl 1.0 g CaCl ₂ · 6H ₂ O 0.1 g MgSO ₄ · 7H ₂ O 2.0 g Na ₂ SO ₄ 1.0 g sodium lactate 2.0 g yeast extract 1.0 g resazurin 1.0 g distilled water 980.0 ml

Solution 2:

FeSO ₄ · 7H ₂ O 0.5 g distilled water 10.0 ml

Solution 3:

sodium thioglycolate 0.1 g vitamin C 0.1 g distilled water 10.0 ml

Solution 4 (trace element solution SL-10):

HCl (25%; 7.7 M) 10.0 ml FeCl ₂ · 4H ₂ O 1.5 g ZnCl ₂ 70.0 mg MnCl ₂ · 4H ₂ O 100.0 mg H ₃ IN ₃ 6.0 mg CoCl ₂ · 6H ₂ O 190.0 mg CuCl ₂ · 2H ₂ O 2.0 mg NiCl ₂ · 6H ₂ O 24.0 mg Na ₂ MoO ₄ · 2H ₂ O 36.0 mg distilled water 990.0 ml

Preparation: all solutions are sterilized separately by autoclaving at 121 ° C for 15 min in a nitrogen atmosphere. Solution 1 is boiled immediately before autoclaving, purging with nitrogen. To prepare the medium, solutions 1, 2 and 3 are mixed, and 1 ml is added from solution 4. The pH of the medium after mixing is set equal to 7.2 ... 7.4.

The disadvantage of this strain is the insufficiently high efficiency of the degree of purification of wastewater from galvanic plants from heavy metal ions.

The objective of the invention is to obtain a highly active strain of sulfate-reducing bacteria used to treat wastewater from heavy metal ions, and thereby expanding the range of bacterial strains used to treat wastewater from heavy metal ions.

The technical result is the use of a strain of bacteria Desulfovibrio sp. SVB-2 for wastewater treatment from heavy metal ions.

The proposed bacterial strain Desulfovibrio sp. SVB-2 is isolated from industrial runoff from galvanic plants containing heavy metal ions. Microorganism strains are highly resistant to heavy metal ions and use organic compounds contained in this waste water as food.

The resulting strain of Desulfovibrio sp. SVB-2 is deposited with the All-Russian Collection of Microorganisms (VKM) of the Institute of Biochemistry and Physiology of Microorganisms named after G.K.Skryabin RAS (IBPM) under registration number VKM V-2285 D.

The proposed strain of Desulfovibrio sp. SVB-2 is not known in science and technology, therefore, the properties that it exhibits are new, and therefore, the claimed solution has an inventive step.

The proposed strain is identified: "Bergey's Manual of Systematic Bacteriology", 1984. VIII ed. v. one; Baltimore-London, William & Wilkins, 964 p .; Bergey's Manual of Systematic Bacteriology, 1986. VIII ed. v.2; Baltimore-London, William & Wilkins, 1599 p .; Bergey's Manual of Determinative Bacteriology, 1989. IX ed. v. 1, Baltimore-London, William & Wilkins, 787 p.

This strain is an active producer of hydrogen sulfide for the precipitation of heavy metal ions from solutions and is characterized by the following cultural and morphological characteristics.

Cultural and morphological characteristics of the patented strain are determined during its cultivation on standard media, the recipes of which are taken from the catalog of the All-Russian collection of microorganisms No. 92 and No. 109 and are given below in the publication "All-Russian collection of microorganisms. Catalog ”, Pushchino-Moscow, VNITI, 1992, 362 p. Cultivation is carried out under anaerobic conditions at 30 ° C for two days.

The proposed strain is characterized by the following features:

The generic and species name of the strain is the bacterial strain Desulfovibrio sp. SVB-2.

Origin - the strain is isolated from the wastewater of galvanic industries.

Morphological characteristic - the shape of the cells is a vibrio, 0.5-1.0 × 3.0 × 5.0 microns in size.

Spore formation - no.

Gram reaction is negative.

Cell motility is motile.

Physiological characteristic - attitude to oxygen - anaerobic.

Growth Eh-0-200 mV. With growth due to anaerobic respiration, electron acceptors: sulfate, sulfite, thiosulfate. Used as substrates in the reduction of sulfates (hydrogen and carbon donors): butyrate, propionate, succinate, lactate, malate, pyruvate. Does not use: acetate, ethanol. It grows at pH 6.5 ... 8.5, the optimum is 7.2. It does not require NaCl for growth, it grows at a temperature of 20-40 °, an optimum of 30 ° C.

Biochemical characteristics of the strain - cells contain cytochrome C and desulfoviridine.

A useful property in connection with which the culture is deposited is the precipitation of heavy metal ions by hydrogen sulfide.

Cultivation conditions - medium used - 63 DSMZ.

Storage conditions - stored by reseeding on the cultivation medium.

An example of the use of a strain of bacteria Desulfovibrio sp. SVB-2 for wastewater treatment from heavy metal ions.

The process is carried out in bioten with loading from pumice with an upward flow of liquid. Seeds are microbial strains grown in a liquid medium in flasks. Biotenes are inoculated with strains of microorganisms Desulfovibrio sp. SVB-2, which are fixed on a pumice stone. The cultivation temperature in the biotank is maintained in the range of 28 ... 30 ° C. Simulated wastewater of galvanic production is passed through the biotank. As a result of the life of the SRB, hydrogen sulfide is released, which combines with heavy metal ions that precipitate.

The efficiency of wastewater treatment containing heavy metal ions is given in the table.

The table shows that the efficiency of wastewater treatment of galvanic production from heavy metal ions when using the proposed strain is 99.9%. Thus, wastewater from galvanic plants can be effectively cleaned of heavy metal ions using the strain Desulfovibrio sp. SVB-2.

Table No. Wastewater Category The concentration of metals, mg / l The sum of heavy metals, mg / l The degree of purification,% Cr Zn Fe Cu Ni COD Wastewater treatment using a prototype strain one Source wastewater 58.8 10.0 115.0 5.2 1,0 650 190.0 - 2 Treated wastewater 0,0 0,0 0.5 0,0 0,0 130 0.5 99.7 Wastewater treatment using the proposed strain 3 Source wastewater 60.0 10.0 118.0 8.9 1,0 530 197.9 - four Treated wastewater 0,0 0,0 0.1 0,0 0,0 75 0.1 99.9

Claims (1)

Bacterial strain Desulfovibrio sp. SVB-2, used for wastewater treatment from heavy metal ions, was deposited by the All-Russian Collection of Microorganisms of the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences (IBPM) under registration number VKM V-2285.