Strain of mycobacterium species smc 65-b, used for purification of fresh and sea water from oil and oil products

 

(57) Abstract:

The invention relates to the field of Microbiology. A new strain of Mycobacterium species (deposited in the Central collection of microorganisms of the State concern "biological product" collectible SMC 65-B) isolated from seawater samples of the White sea. Strain suitable for purification of fresh and sea water (up to 7.5% salinity) from pollution by oil and oil products in a wide range of temperatures (10-26oC). The technical result of the invention is the expansion of the scope and efficiency of purification from oil pollution. 3 table.

The invention relates to the field of Microbiology and the receipt of a new strain of bacteria, it is suitable for cleaning fresh and marine waters from pollution by oil within 7-14 days in a wide range of temperatures (10-26oC).

A known strain of Pseudomonas putida 36 (1), which has a high capacity for disposal of oil and oil products.

However, the disadvantage of this strain is the inability to grow and to dispose of the oil at low temperatures. Strain Ps. putida 36 is not suitable for the cleaning of oil polluted sea water.

A known strain of Mycobacterium flavescens (2) used for water purification and soil from is the inability to clean up oil pollution of sea water.

Known consortium of microorganisms called Devouroil (3) that allows you to clean from oil pollution water with a high % salinity (up to 10%). The consortium works effectively when the temperature of the 28oC.

The disadvantage of this consortium is the lack of ability to clean oil spills at low temperatures.

The objective of the invention is the obtaining of a bacterial strain carrying out the purification of sea water and fresh water from oil pollution in a wide range of temperatures (10-30oC) in a relatively short time.

The proposed strain of Mycobacterium sp. isolated from seawater samples of the White sea (the waters of the cargo port of Arkhangelsk). The strain deposited with the Central collection of microorganisms of the State concern "biological product", collection number SMC 65-B.

In experiments on white mice strain is not virulent and non-toxic.

The resulting strain develops from seed mineral environment, using as a source of carbon hydrocarbons of oil and oil products, and is characterized by the following morphological, cultural and physiological characteristics.

Morphological features.

In a visible gram nepalse grammaticale. On meat-peptone agar in a day, there are small round smooth edge of the colony color of melted milk. Colonies are opaque, shiny, oily, easily removed from the agar. For 3-4 days along with a smooth appear rough (3-4%) colonies. In meat-peptone broth culture gives a uniform turbidity and sediment at the bottom.

Physiological and biochemical characteristics.

Mycobacterium sp. SMC 65-B is a facultative aerobe in a wide temperature range 4 - 37oC. From carbohydrates sprayway glucose, sucrose, mannose, arabinose, sorbitol, dulcet, xylose, galactose, mannitol. Starch is not hydrolyses. Proteolytic activity is not. Strain catalanopolonesa, oxidization; has urease activity, nitrate does not recover, phenylalanine and will not desaminase. Additional growth factors is not needed.

The culture is stored in lyophilized form in ampoules at 4oC. the viability of the strain can be maintained by regular re-seeding (1 every 2 weeks) on the mineral environment 8E with oil and a full environment.

The ability of the proposed strain to the biodegradation of oil was studied in relation to marine and fresh water, as svstem Mycobacterium sp. SMC 65-B grown in 250 ml flasks containing 50 ml of sterile medium. The environment is prepared with fresh water containing 0.75 g/l disubstituted ammonium phosphate. Sterile petroleum products contribute by weight with an accuracy of up to 0.05 mg. the substrate Concentration is 0.5% and 1%. Environment infect a two-day agar culture at the rate of 107cells/ml. Flask exhibit in stationary conditions at 26 and 10oC for 14 days, as well as on the rocking chair (150 rpm) at 26oC for 7 days. As a control using uninfected bulb with medium and petroleum products. After a period of incubation the percentage of recycled substrate determined by the gravimetric method after extraction of petroleum chloroform. These experiments are presented in table. 1.

Disposal of oil at an initial level of contamination of 0.5% in the stationary conditions 26oC is 48%. Lowering the temperature to 10oC leads to a decrease of the level of treatment to 39%. Aeration in the rocking chair at the 26oC increases the rate of recycling in 7 days (versus 14 days in stationary conditions) cleaning reaches 48%.

Disposal diz. fuel in fresh water at an initial level of contamination of 1% in a hospital for 14 days at 26oC SOS the fuel, after 7 days the level of treatment is 67%.

Example 2. Dispose of oil in sea water.

Strain of Mycobacterium sp. grown in 250 ml flasks containing 50 ml of sterile medium. The environment is prepared with sea water (3.5% of sea salt) containing 0.75 g/l disubstituted ammonium phosphate. Sterile petroleum products contribute by weight with an accuracy of up to 0.05 mg Concentration of the substrate is 0.5% oil and 10% diz. fuel. Environment infect a two-day agar culture at the rate of 107cells/ml. Flask exhibit in the stationary conditions 26oC and 10oC for 14 days, as well as on the rocking chair (150 rpm) at 26oC for 7 days. As a control using uninfected bulb with medium and petroleum products. After a period of incubation the percentage of recycled substrate determined by the gravimetric method after extraction of petroleum chloroform. Data of the experiment are presented in table. 2.

Recycling of oil in sea water at an initial level of contamination of 0.5% in a hospital for 14 days at 26oC is 44.6% and hardly decreases when the temperature decreases to 10oC (42.6%). Additional aeration in the rocking chair at the 26oC leads to faster recovery and stand the Disposal of diz. fuel in sea water at an initial level of contamination of 1% in a hospital for 14 days at 26oC is 45,6%, with 10oC was 47.1%. Additional aeration in the rocking chair at the 26oC increases the rate of utilization of diz. fuel: after 7 days the level of treatment is 82%.

Example 3. Recycling of oil in sea water with different amounts of sea salt.

Strain of Mycobacterium sp. grown in 250 ml flasks containing 50 ml of sterile medium. The content of sea salt in the environment varies from 0 to 7.5%. As an additional mineral foundations make the disubstituted ammonium phosphate based 0.75 g/l Concentration of oil is 0.5%. Environment infect a two-day agar culture at the rate of 107cells/ml. Flask exhibit in a hospital for 14 days at 26oC. as a control using uninfected bulb with medium and oil. After a period of incubation the percentage of recycled substrate determined by the gravimetric method after extraction of oil chloroform. Data of the experiment are presented in table. 3.

As can be seen from the table, the strain carries out the disposal of oil even at high salinity (7.5 percent) of water. The maximum level of bioremediation (56%) will designsthere ability to bioremediation (50-82%) of marine and fresh water from oil pollution in a wide range of temperatures (10-26oC) in a relatively short time.

The proposed strain differs from the prototype in that it keeps the ability to biodegradation of crude oil in salt water when the temperature decreases from 26 to 10oC. in Addition, Mycobacterium sp. SMC 65-B compared to the prototype is developing fine and recycles oil in fresh water in the temperature range 10-26oC.

The above data suggest that strain of Mycobacterium sp. SMC 65-B for treatment of fresh and marine waters from oil pollution.

Sources of information

1. Auth. St. USSR N 1076446, C 12 N 15/00, 1984, bull. N 8.

2. Patent RU N 2053296, C 12 N 1/20, 1996, bull. N 3.

3. Patent RU N 2023686, CL 02 F 3/34, 1994, bull. N 22.

Strain of Mycobacterium species SMC 65-B deposited in the Central collection of microorganisms of the State concern "biological product" used for purification of fresh and sea water from oil and oil products in a wide range of temperatures.

 

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2 dwg, 12 ex

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