Quality assessment method of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys in operation of spacecrafts, and suspension of spore materials for its implementation

FIELD: biotechnology.

SUBSTANCE: method of qualitative assessment of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys in operation of spacecrafts and the suspension of spore materials fungi of implementation of the said method is proposed. The test and control samples of aluminium and magnesium alloys are prepared. The prepared samples are dried, sterilised. Fungal cultures - strains of microorganisms Paecilomyces variotii Bainier of All-Russian collection of microorganisms F-4039D, Ulocladium botrytis Preuss of All-Russian collection of microorganisms F-4032D, Penicillium chrysogenum Thorn of All-Russian collection of microorganisms F-4034D, Aspergillus sydowii (Bainier et Sartory) Thorn et Church of All-Russian collection of microorganisms F-4037D, Cladosporium sphaerospermum Penz. of All-Russian collection of microorganisms F-4041D are inoculated for growing spores into tubes with a sloped agar medium of Czapek-Dox. The tubes are thermostated at a temperature of (29+2)°C for 14-28 days until appearance of mature spores. Spore materials suspensions of individual cultures of the said fungi are prepared, which are then mixed in equal proportions. And the concentration of spores of each fungi species in the suspension should be in the range of 1-2 mln/cm3. The resulting suspension is applied to the sterilised test samples. Then they are dried and placed as one sample in each Petri dish on the surface of agar medium of Czapek-Dox. The control samples not treated by the spore material are also placed on the surface of the agar medium of Czapek-Dox in the Petri dishes. The Petri dishes with the test and control samples are placed in different desiccators at the bottom of which water is poured to maintain the humidity more than 90%. The desiccators are closed and incubated in the thermostat at a temperature of (29±2)°C. The exposition of test and control samples is carried out for 40, 120 and 180 days. Then the samples are removed from the Petri dishes, and the corrosion products and mycelium are removed from the surface of the samples washing them in running water. The samples are soaked in 70% ethyl alcohol for 30 minutes, then they are washed with detergent and dried. Then, using a scanning electron microscope a qualitative assessment of biocorrosion damages is carried out. According to the change in the appearance of the surface of the samples the initial stages of biocorrosion and its type is evaluated, as well as the distribution of corrosion damage on the surface of the samples, and the dependence of the biocorrosion process on time is determined.

EFFECT: inventions enable to carry out the qualitative assessment of the initial stages of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys with the thickness of more than 2 mm.

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The invention relates to the technical Microbiology and biocorrosion tests, and in particular to methods of determining exposure to aluminum-magnesium alloys used in aerospace technology, to the impacts technophilic strains of microorganisms isolated in real operating conditions with the structural elements of the Russian segment (PC) of the International space station (ISS) and deposited in the all-Russian collection of microorganisms (VKM).

Space station - the high risk area with extreme operation conditions of the equipment. Constant monitoring of all parameters of the internal environment, including flora station, provides improved reliability and security of its operation.

All materials in the process of operation will be subject to various environmental influences that cause corrosion processes on their surface. They can be caused by exposure to chemicals (chemical corrosion), and the action of various microorganisms (biological corrosion). Real corrosion processes typically occur under the action of both factors.

The action of microorganisms is one of the factors contributing to the emergence and development of corrosion processes of metals (Albitskaya O.N., Shaposhnikova N.A. "Influence Plesen the th on the corrosion of metals" // Microbiology. 1960. So 29. S-730; Koval ET, Kasyan D.M., Danowski VI "Studies of fungal corrosion" // Biological damage of building and industrial materials. Kiev: Naukova Dumka, 1978, p.59-60; Lugauskas, A., Mikulskis A.I., Laurene DU Directory of micromycetes-biodestruction polymeric materials. Resp. edit Mwhalen, M.: Nauka, 1987, 340 S.).

In the basis of the action of microorganisms on metals lies electrochemical mechanism. Microorganisms can cause or modify the corrosion in three main ways:

1. Directly acting on the kinetics of electrode reactions.

2. The formation of metabolites with corrosive properties (inorganic and organic acids and the like).

3. Changes on the boundary surface of the metal-electrolyte interface, which can lead to corrosion (for example, the formation of areas with increased formation of oxides) (Costello J.A., 1969. The corrosion of metals by micro-organisms. Int. Biodent. Bull., 5, 101).

Fungi develop on surfaces in contact with metals (fabric, paint, fuel, etc.), where spores are spread by condensation of moisture begin to develop, forming organic acids.

Corrosion promotes condensation of the water vapor in the mycelium of the fungus, the accumulation of them in the growth process of organic acids. In addition, under the mycelium of fungi are conditions, b is appopriate for the development of other microorganisms.

Aluminum alloys are widely used in the aerospace industry, shipbuilding. During the long term stability and reliability of components and subassemblies largely depends on the processes of corrosion, including corrosion caused by microorganisms, bio - corrosion. In the basis of the action of microorganisms on metals lies elektrohimicheskiy mechanism. Microorganisms can cause corrosion damage and strengthen them, directly affecting the kinetics of electrode reactions.

A known method for determining indicators biocorrosion damage steels, aluminum oxide and magnesium alloys (patent RU №1766073, 1995, IPC6: C12Q 1/02, C12Q 1/18), characterized in that the prepared samples metals is dried in a drying Cabinet at a temperature of 80°C for 15 minutes, weighed, wrapped in Kraft paper, sterilized in an oven for 2 hours at a temperature of 165°C, then incubated the samples on the surface of the lawn test-cultures grown on nutrient medium wort agar in a Petri dish. On the surface of the medium wort agar in a Petri dish placed one bacteriological loop spores of the fungus Aspergillus flavus Link BKMF-3190 D and RUB evenly with a spatula over the entire surface of the nutrient medium in Petri dishes wrapped in paper and termostatico is at a temperature of 30°C. After incubation, when the medium surface to form a continuous growth of the fungus on its surface with sterile forceps have sterilized samples of different metals. The number of samples tested each metal must be at least 5 pieces of Control are the same samples of metals, but is placed on the surface of a sterile nutrient medium in Petri dishes, cropped the test culture. Prepared for the tests in Petri dishes with a metal sample is placed in the desiccator, the bottom of which is filled with sterile distilled water, resulting in a desiccator maintain humidity 96-98%. Executory placed in a thermostatic room at a temperature of (30±2)°C. the Duration of exposure the test and control samples 9 months. After finishing the test Petri dishes with metallic samples subjected to autoclave sterilization VK-75 at a pressure of 2 ATM and a temperature of 120°C 2 hours After sterilization of metal samples purified from the mycelium, medium and corrosion products with a scalpel. The products of corrosion of aluminum alloys is removed in solution composition: chromium (VI) oxide 20 g/DM3; orthophosphoric acid (pl. 1,59) 50 cm3/DM3; the temperature of 80-95°C.

When removing corrosion product samples with tweezers in a vertical position immersed in the solution and allowed to stand for 1 min For the em specimens are removed from the solution, washed first in water, then in acetone, dried and weighed.

The weight loss of the samples is determined by the formula

K15Δm15mo100%

where Δmthe mass loss of the test sample;

Δm, mo, m, where mois the mass of the sample before testing, g;

m is the mass of the sample after testing and removal of corrosion products,

The disadvantage of the prototype method is the inability to evaluate the initial stages biocorrosion damage due to low sensitivity of the proposed method. Therefore, commonly used to assess the extent of corrosion weighing samples, as in the prototype, does not give clear results at the initial stages of the process of bio-corrosion, when weight loss is negligible. In addition, when the weighing is impossible to judge about the nature of the damage caused by the used microorganisms.

Known sham Penicillium chrysogenum BKMF-3068D used as the test culture for testing aluminum alloys for funginertness (inventor's certificate SU # 1606530, 15.11.1990, IPC 5: C12N 1/14, C12Q 1/02).

A known strain of the fungus Aspergillus flavus Link as the test culture to determine grebastica steels, aluminum oxide and magnesium alloys (patent RU №1766073, 1995, IPC6: C12Q 1/02, C12Q 1/18), taken as a prototype.

The disadvantages of these strains is their low activity with respect to aluminum-magnesium alloys used in hermozamente volumes, a feature of which is the presence of harmful trace (VMP), which are specific nutrient medium, forming biological aggressiveness (organic compounds with high biocorrosion activity) in respect of various materials including aluminum-magnesium alloys.

The use of fungi from the collection of the ACM in accordance with GOST 9.048-89, as a rule, does not give the expected results, because the culture of fungi during storage and numerous subcultures lose activity. The use of the method in the frequency of occurrence of species for assessment of biodeterioration of materials more revealing than the traditional method of scoring. Mushrooms, featured GOST 9.048-89 testing funginertness not correspond to the actual prevalence materials. Therefore, it is sensible to use the same set of mushrooms, which have a fairly large frequency of occurrence directly on your mater is ale (Journal Mycology and Phytopathology", volume 31, issue 2, Saint-Petersburg, 1997, p.85).

Thus, the well-known collection strains of microorganisms may not be used for qualitative assessment biocorrosion lesions thin-walled hermetic shell of aluminum-magnesium alloys.

Object of the invention is a qualitative assessment of the initial stages biocorrosion lesions thin-walled hermetic shell of aluminum-magnesium alloys, of a thickness not exceeding 2 mm with different kinds of surface treatment (machined, chemically polished, anodized), modeled depending on the operational conditions (location of the material in the operating pressure chamber), and the selection of the consortium of strains of microorganisms, naturally formed depending on the specific location of structural elements in terms hermosanuevo volume.

The technical result of the invention is:

detection of the initiation of the process of bio-corrosion, definition biocorrosion effects (the evaluation of the appearance of the affected surface - determine the type of corrosion, the distribution of corrosion damage and the characteristic of its form, different from chemical corrosion; dependence biocorrosion process from time to time);

- the possibility of increasing the reliability and life of constructio the materials in the presence of technophilic microorganisms in operating conditions hermosanuevo volume.

Declared suspension of spore material including a mixture of spore-bearing materials from individual cultures of fungi Paecilomyces variotii Bainier VKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thom BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D, is a promising test when testing aircraft design materials on biocorrosion resistance.

The technical result is achieved by a method qualitative assessment biocorrosion lesions thin-walled hermetic shell of aluminium-magnesium alloys with the operation of spacecraft is characterized by the fact that made the test and control samples of aluminum-magnesium alloys, prepared the samples dried, sterilized, for growing culture of the fungi spores inoculated in tubes with beveled agar medium of čapek-doksa, thermostatic tubes at a temperature (29±2)°C for 14-28 days before the appearance of Mature spores, preparing a suspension of spore material, which is applied to sterilized test samples, then dried them and put one sample in Petri dishes on agar surface environment of čapek-doksa, control samples not treated spore material, also feature on the surface of the agar medium of čapek-doksa in Petri dishes, Petri dishes with and the probation and control samples are placed in different executory, at the bottom of which pour water to maintain humidity over 90%, executory closed and incubated at a temperature (29±2)°C, conducting exposure of the test and control samples, then the samples are then removed from the Petri dishes, remove the corrosion products and the mycelium from the sample surface, rinsing them under running water, identify indicators biocorrosion lesions on the environment of čapek-doksa prepare a spore suspension of materials from individual cultures of fungi - bacteria strains of Paecilomyces variotii Bainier BKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thom BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D, which are then mixed in equal proportions, and the concentration of spores of each species of fungus in the suspension should be in the range of 1-2 million/cm3obtained suspension by means of a spray is applied on the test samples of aluminum-magnesium alloys with different kinds of surface treatment - machined, chemically polished, anodized, after washing the samples in flowing water, they kept in 70%ethyl alcohol for 30 minutes, then washed with detergent and dried, then using a scanning electron microscope conduct a qualitative assessment biocorrosion lesions: change the appearance of the surface of the samples began to appreciate the major stages of bio-corrosion and its type, the distribution of corrosion damage on the sample surface, and determine the dependence biocorrosion process from the time when the exposure time of the test and control samples performed within 40, 120 and 180 days.

The technical result is achieved by using spore suspensions of materials, including a mix of spore-bearing materials from individual cultures of fungi Paecilomyces variotii Bainier BKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thom BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D in equal proportions, to assess biocorrosion lesions thin-walled hermetic shell of aluminium-magnesium alloys with the operation of spacecraft.

The microorganism strain Paecilomyces variotii Bainier BKM F-4039D.

The property of the microorganism. The ability to cause damage of structural materials used in aerospace engineering (aluminum-magnesium alloys, polymeric materials), the origin of hermosanuevo volume of the spacecraft.

Pedigree strain (site selection, source selection, and other information about the movement of the strain). The strain isolated from the dust collected by the air filter on the Russian Segment of the International Space Station (PC MKC) in the period of the ISS-9, October 24, 2004

The method of obtaining strain. Sowing was carried out on solid nutrient cf the control method of fingerprint fragment filter.

The authentication method determiner, which was carried out identification. Identification based on morphological characters. Keys: R.A. Samson: Paecilomyces and some allied Hyphomycetes. // Studies in Mycology. 1974. No. 6. 119 pp.

Cultural-morphological and microscopic characteristics of the strain. Colonies on the environment wort agar (malt-agar growing very quickly and reach a diameter of 8 cm for 7 days at 25°C. Velvety pubescence flat, sometimes fluffy and then felt quite high, very small. Consist of a dense layer conditoner structures. The mycelium is colorless. Sporulation from dark Krasnogo to olive-beige-brownish color. Exudate absent or inconspicuous. The reverse of the colony yellowish to yellow-brown and almost black, with abundant formation of chlamydospores. Conidiophores smooth, slightly rough, or with inlay, are quite dense brush, consisting of disordered twigs, bearing Metula with 2-7 Felidae. Basically 35-90×3.5-7.0 µm, but sometimes up to 150 µm in length. Tiality in bunches or single, variable in shape and size, 12-20(35)×2.5-5.0 µm, have a cylindrical abdomen and long, thin neck. Conidia hemispherical, club-shaped, elliptic, smooth, 3.2-15.0×2.0-5.0 µm. Chlamydospores are usually solitary or in short chains, rounded, thick-walled, dark-colored, gadkari slightly rough 4.0-8.0 (10) µm in diameter.

Description of the conditions necessary for the cultivation of the strain (medium composition, temperature, time of cultivation, and so on). The cultivation conditions of strain features are not standard for most filamentous fungi.

Environment:

- Chapek's medium with 3%sucrose content (sucrose 30 g; NaNO32 g; KH2PO41 g; KCl, 0.5 g; MgSO47H2About - 0.5 g; FeSO47H2O - 0.001 g; agar 20 g; tap water - 1 liter);

- wort-agar (16 - ballymaloe nikolenka beer wort - 150 ml; agar 20 g; tap water is 800 ml); the temperature of 22-25°C; term cultivation 7-10 days.

The optimum pH of cultivation: 6-6,5.

Description storage mode strain (environments, conditions, time limit, etc.). For keeping fit all the methods. The strain was maintained through periodic transfers on the environment wort agar.

This filamentous fungus is not listed as pathogenic in the "Regulations on the procedure for accounting, storage, handling, dispensing and delivery of cultures of bacteria, viruses, Rickettsia, fungi, protozoa, Mycoplasma, bacterial toxins, poisons of biological origin".

The strain of microorganism Ulocladium botrytis Preuss BKM F-4032D.

The property of the microorganism. The ability to cause damage of structural materials used in aerospace engineering (aluminum-magnesium alloys, polymeric materials), and the origin is of an unforgettable - from hermosanuevo volume of the spacecraft.

Pedigree strain (site selection, source selection, and other information about the movement of the strain). The strain isolated from the dust collected by the air filter on the Russian Segment of the International Space Station (PC MKC) in the period of the ISS-8, April 30, 2004

The method of obtaining strain. Sowing was carried out on solid medium method of fingerprint fragment filter.

The authentication method determiner, which was carried out identification. Identification based on morphological characters. Keys: Ellis MV More Dematiaceous Hyphomycetes. Commonwealth Mycologigical institute, Kew., Kew: Surrey, 1976. Simmons E.G. Typification of Altemaria, Stemphylium and Ulocladium // Mycologia. 1967. Vol.59. Page 67-92.

Cultural-morphological and microscopic characteristics of the strain. Colonies on potato-dextrose and wort-agar growing reach 5-6 cm in diameter for 7 days. Velvety, slightly small, powdery, can be slightly fluffy. Olive-brown to black color. On the environment of čapek colony more limited. Conidiophores erect or prostrate, short, highly branched, crankshaft-curved with conidia formed in the nodes. The colour ranges from almost unpainted to brown, conidial scar remaining around the pores colorless. The size of conidiophores up to 50×4.5 µm. Conidia Moralny (mnogolet the data with longitudinal and transverse septa are formed of a single, very rarely in short chains, ovoid form, without spout, first, light brown, olive, rapidly darkening to almost black, the surface is rough warty until lumpy. The sizes vary within 19-25×7-12 µm, with (1-)2-3 transverse and 0-2 oblique or longitudinal septa. Secondary conidiophores are formed extremely rare.

Description of the conditions necessary for the cultivation of the strain (medium composition, temperature, time of cultivation, and so on). The cultivation conditions of strain features are not standard for most filamentous fungi.

Environment:

- Chapek's medium with 3% sucrose (sucrose 30 g; NaNO32 g; KH2PO41 g; KCl, 0.5 g; MgSO47H2O - 0.5 g; FeSO47H2About - 0.001 g; agar 20 g; tap water - 1 liter);

- wort-agar (16-ballymaloe not hopped beer wort - 150 ml; agar 20 g; tap water is 800 ml); the temperature of 22-25°C; term cultivation 7-10 days.

The optimum pH of cultivation: 6-6,5.

Description storage mode strain (environments, conditions, time limit and so on). For keeping fit all the methods. The strain was maintained through periodic transfers on the environment wort agar.

This filamentous fungus is not listed as pathogenic in the "Regulations on the procedure for accounting, storage, handling, dispensing and delivery of cultures of bacteria, viruses, Rickettsia, fungi, protozoa, Mycoplasma, bacterial toxins, poisons of biological origin".

The strain of microorganism Penicillium chrysogenum Thom BKM F-4034D.

The property of the microorganism. The ability to cause damage of structural materials used in aerospace engineering (aluminum-magnesium alloys, polymeric materials), and the origin of hermosanuevo volume of the spacecraft.

Pedigree strain (site selection, source selection, and other information about the movement of the strain). The strain isolated from the dust collected by the dust collector on the Russian Segment of the International Space Station (PC MKC) in the period of the ISS-9, October 24, 2004

The method of obtaining strain (mutant). Planting on a solid nutrient medium from the suspension of dust.

The authentication method determiner, which was carried out identification. Identification based on morphological characters. Keys: Raper K.B., Thom S. Manual of the Penicillia. New York: Heftier Publishing Co. 1968. 875 p. Ramirez C. Manual and Atlas of the Penicillia. Amsterdam; New York; Oxford: Elsiveier Biomedical Press, 1982. 874 p. Pitt G.I. A laboratory guide to common Penicillium species (sec. ed.). North Ryde, U.S.W., Australia: CSIRO, Division of Food Processing, 1991. 188 p.

Cultural-morphological and microscopic characteristics of the strain. Colonies growing on the Chapek's medium with 3% sucrose, 7 days growth at 25°C, 35-50 mm in diameter. The surface is velvety, the colony radial is folded, dull green mass of conidia form characteristic of the crust, the exudate is usually formed mycelium colorless. The reverse side unpainted, cream or yellow-brown. On wort-agar colony growing more velvety. Conidiophores aggregated in dense fascicles on substrate mycelium, rough rough, 200-400 µm in length. Brush usually with a three-tiered pressed twig compact, Metulla on 3-5 verticil 10-15×3-3,5 µm, are 5-8 butylamine of failed 9-12×2.5-3 µm. Conidia are almost spherical, 3.5-4.0 µm.

Description of the conditions necessary for the cultivation of the strain (medium composition, temperature, time of cultivation, and so on). The cultivation conditions of strain features are not standard for most filamentous fungi.

Environment:

- Chapek's medium with 3% sucrose (sucrose 30 g; NaNO32 g; KH2PO41 g; KCl, 0.5 g; MgSO47H2About - 0.5 g; FeSO47H2O - 0.001 g; agar 20 g; tap water - 1 liter);

- wort-agar (16-ballymaloe not hopped beer wort - 150 ml; agar 20 g; tap water is 800 ml); the temperature of 22-25°C; term cultivation 7-10 days.

The optimum pH of cultivation: 6-6,5.

Description storage mode strain (environments, conditions, time limit and so on). For keeping fit all the methods. The strain was maintained through periodic transfers on the environment wort agar.

Given the initial view of the filamentous fungus is not listed as pathogenic in the "Regulations on the procedure of accounting, storage, handling, dispensing and delivery of cultures of bacteria, viruses, Rickettsia, fungi, protozoa, Mycoplasma, bacterial toxins, poisons of biological origin".

The strain of microorganism Aspergillus sydowii (Bainier et Sartory) Thom et Church BKMF-4037D.

The property of the microorganism. The ability to cause damage of structural materials used in aerospace engineering (aluminum-magnesium alloys, polymeric materials), and the origin of hermosanuevo volume of the spacecraft.

Pedigree strain (site selection, source selection, and other information about the movement of the strain). The strain isolated from the dust collected by the dust collector on the Russian Segment of the International Space Station (PC MKC) in the period of the ISS-9, October 24, 2004

The method of obtaining strain (mutant). Planting on a solid nutrient medium from the suspension of dust.

The authentication method determiner, which was carried out identification. Identification based on morphological characters. Keys: Raper K.B., Fennel D.I. The genus Aspergillus. Baltimore: Williams & Wilkins, 1965. 686 p. M.A. Klich Identification of common Aspergillus species. CBS 2002. 116 R.

Cultural-morphological and microscopic characteristics of the strain. Colony on Chapek's medium at 25°C are growing well, reaching 3-4 cm in diameter for 10 days, folded in the center raised. The structure of the colony dense velvety, mycelium be the first, the color of the visible blue-green exudate usually rich, straw-yellow to dark red-brown. The reverse side of the colony from colorless to coral-pink, maroon or reddish-black with aging. On the environment wort agar colonies growing more flat velvety. Conidial heads are radial to 100-150 µm in diameter. Conidiophores hyaline, thick-walled, smooth, up to 500 µm in length. From aerial mycelium leave a short conidiophores with partially reduced conidiogenesis structures. Bubble rounded up to 20 µm in diameter. Heads biseriate, Metulla 6-7×2-3 µm, Fieldy 7-10 x 2-2 .5 µm. Conidia spherical, considerably prickly, 2.5-3(3.5) µm. Sometimes formed vitreous cells rounded.

Description of the conditions necessary for the cultivation of the strain (medium composition, temperature, time of cultivation, and so on). The cultivation conditions of strain features are not standard for most filamentous fungi.

Environment:

- Chapek's medium with 3% sucrose (sucrose 30 g; NaNO32 g; KH2PO41 g; KCl, 0.5 g; MgSO47H2O - 0.5 g; FeSO47H2O - 0.001 g; agar 20 g; tap water - 1 liter);

- wort-agar (16-ballymaloe not hopped beer wort - 150 ml; agar 20 g; tap water is 800 ml); the temperature of 22-25°C; term cultivation 7-10 days.

The optimum pH of cultivar is for: 6-6,5.

Description storage mode strain (environments, conditions, time limit, etc.). For keeping fit all the methods. The strain was maintained through periodic transfers on the environment wort agar.

This filamentous fungus is not listed as pathogenic in the "Regulations on the procedure for accounting, storage, handling, dispensing and delivery of cultures of bacteria, viruses, Rickettsia, fungi, protozoa, Mycoplasma, bacterial toxins, poisons of biological origin".

The microorganism strain Cladosporium sphaerospermum Penz. BKM F-4041D.

The property of the microorganism. The ability to cause damage of structural materials used in aerospace engineering (aluminum-magnesium alloys, polymeric materials), and the origin of hermosanuevo volume of the spacecraft.

Pedigree strain (site selection, source selection, and other information about the movement of the strain). The strain isolated from the dust collected by the dust collector on the Russian Segment of the International Space Station (PC MKC) in the period of the ISS-11, October 15, 2005

The method of obtaining strain (mutant). Planting on a solid nutrient medium from the suspension of dust.

The authentication method determiner, which was carried out identification. Keys: Ellis MV Dematiaceous Hyphomycetes. Commonwealth Mycologigical institute, Kew., Kew: Surrey, 1971. 609 p. Zalar P, Hoog GS de, Schroers H-J, Crous PW, Groenewald JZ, Gunde-Cimerman N (2007). Phylogenyand ecology of the ubiquitous saprobe Cladosporium sphaerospermum, with descriptions of seven new species from hypersaline environments. Studies in Mycology 58:157-183.

Cultural-morphological and microscopic characteristics of the strain. Colonies on potato-dextrose agar at 25°C on day 7 reach 21-44 mm in diameter. Velvety, smooth or wrinkled, dark-olive-brown-black. The exudate is not formed, the reverse is brown-black. Mycelium dense dark-colored aerial mycelium is usually absent. Conidiophores poorly branching upright, olive-brown, smooth or gently rough, with several extended top with dark scars from conidia length is highly variable (10-)45-130(-300)×(2.5-)3-4(-6) µm. Conidial chain branched tree. Ramachandi with 2-3 septa, conidia following orders from 1-0 septa terminal conidia ovoid or subspherical, smooth or gently warty, prostrannye to both ends with a noticeable scar. The size of the terminal conidia (2.5-)3-4(-7)×(2-)3-3 .5(-4.5) µm. Ramachandi second-order cylindrical (4-)9-17 .5(-48.5)×(2-)3-3 .5(-8.5) µm.

Description of the conditions necessary for the cultivation of the strain (medium composition, temperature, time of cultivation, and so on). The cultivation conditions of strain features are not standard for most filamentous fungi.

Environment:

- Chapek's medium with 3% sucrose (sucrose 30 g; NaNO32 g; KN2PO4 1 g; KCl, 0.5 g; MgSO47H2O - 0.5 g; FeSO47H2O - 0.001 g; agar 20 g; tap water - 1 liter);

- wort-agar (16-ballymaloe not hopped beer wort - 150 ml; agar 20 g; tap water is 800 ml); the temperature of 22-25°C; term cultivation 7-10 days.

The optimum pH of cultivation: 6-6,5.

Description storage mode strain (environments, conditions, time limit and so on). For keeping fit all the methods. The strain was maintained through periodic transfers on the environment wort agar.

This filamentous fungus is not listed as pathogenic in the "Regulations on the procedure for accounting, storage, handling, dispensing and delivery of cultures of bacteria, viruses, Rickettsia, fungi, protozoa, Mycoplasma, bacterial toxins, poisons of biological origin".

The above strains of microorganisms: Paecilomyces variotii Bainier BKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thom BKM F-4032D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D deposited by the all-Russian collection of microorganisms, Institute of biochemistry and physiology of microorganisms, name Gscreen (142290, Pushchino, Moscow region, Prospekt nauki, 5).

The invention consists in the following.

The method is based on testing and application of a suspension of spores of fungi on the surface of samples of aluminum-magnesium alloys taken the methods described in GOST is x 9.048-89 and 9.049-91. Tests biocorrosion lesions is carried out in an accelerated mode, which is characterized by elevated temperature (29±2)°C and 90%humidity, for 40, 120, 180 days, when the influence of these factors. Testing is carried out in a humid chamber (desiccator).

For testing use two sampling: test and control. The test sample is designed to determine the intensity of the development of fungi and their effect on the product parameters, a control sample to determine the effect on the parameters of products of high humidity and high air temperature without the action of fungi, to compare the test results with the test sample. Test and control samples made from a cylindrical billet of the material of construction is aluminum-magnesium alloy with a thickness of 5 mm and a diameter of 16 mm (GOST/TU - 1-90395-91, melting batch No. 89606) with different kinds of surface treatment: machined, chemically polished, anodized.

Prepared samples are dried and sterilized at a temperature of 165°C for 2 hours.

For growing dispute culture fungi strains of microorganisms Paecilomyces variotii Bainier BKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thom BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D - inoculated in tubes with RMSE of the military agar medium of čapek-doksa, then thermostatic tubes at a temperature (29±2)°C for 14-28 days before the appearance of Mature spores. Culture of fungi are supported on the environment wort agar.

For the preparation of a suspension of spores obtained using 14-28-day cultures of the fungi. The spore suspension is prepared separately for each species of fungus with the use environment of čapek-doksa. To do this in a test tube or flask containing 25 cm3environment of čapek-doksa, carry spores from the surface of the grown culture through the loop. As the liquid dispersion medium water use. A suspension of each strain of the fungus is prepared separately: Paecilomyces variotii Bainier BKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thom BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D. The suspension is thoroughly shaken and filtered through 4 layers of sterile gauze. The concentration of spores of each fungus counted separately in the Goryayev camera or photoelectric colorimeter. To determine the concentration of spores using a photoelectric colorimeter using a filter that creates a wavelength λ=400 nm, and the cell (50±0,5) mm Optical density, corresponding to a concentration of 1-2×106/cm3for Paecilomyces variotii, 0,28 is 0.55. The density of the suspensions of other strains evaluated using the camera Goryaeva, due to the nature of the spores of these species.

A spore suspension of kadogawa mushroom mixed in equal volumes and used to infect samples of metal. The suspension is applied to a sterile test samples evenly by means of a spray (e.g., sprinklers). The thus treated samples kept in a laminar box at a temperature of 25°C before drying. The suspension is applied from two sides that are subsequently dried in a laminar box. Control samples treated with sterile environments and as a test, placed on the surface of sterile agar medium of čapek-doksa in Petri dishes, which are then put in executory.

Then the samples contaminated metal (test samples) are placed in the centre of Petri dishes (one sample per Cup), pre-filled agar medium of čapek-doksa. Petri dishes with test and control samples are placed in different executory, the bottom of which water is poured to create 90%humidity, and maintained at a temperature (29±2)°C to monitor the initial stages biocorrosion damage. Every 7 days executory reveal for 3 minutes for fresh air. The time of incubation, 40, 120 and 180 days.

After the completion of incubation, the samples removed from the Petri dishes, and the resulting mycelium and corrosion products are removed from the sample surface (in accordance with ISO 8407) by easy mechanical cleaning using a soft brush with subsequent interest is affected by washing in running water. Then the samples are incubated in 70%ethyl alcohol for 30 minutes, then washed with detergent (e.g., "Fairy Plus, Procter & Gamble and again running tap water followed by rinsing in distilled water. After that, the samples are dried in Boxing. Control samples subjected to the same treatment. Experiments carried out in triplicate.

Test and control samples stored in sterile Petri dishes before the examination in a scanning electron microscope.

In the experiments, there was complete fouling samples of mycelium in the presence of mineral medium with sucrose, which is the carbon source for fungal growth and contributing to the formation of acidic products of their activity. Thus, there is the possibility to investigate the direction and nature of damage to aluminum-magnesium alloys under the influence of such factor as the fouling that can occur in terms of violations of temperature and humidity conditions (higher temperature and humidity) in the life-support system in hermozamente volumes.

The dried samples are examined using a scanning electron microscope (SEM)used for observation of the microrelief of various surfaces.

Microscopic examination of the samples was held the but on scanning electron microscope SCAN ITSELF firm CAMBRIG (accelerating voltage 20 kV, the registration mode is a secondary electron image). The alloy samples, which could not stay for organic compounds, were placed in the microscope without pre-treatment, and samples from the remains of organisms previously deposited alloy of gold and palladium by the method of ion sputtering of the metal in an argon atmosphere (the layer thickness of 25 nanometers).

The invention is illustrated by tables.

In tables 1, 5, 9 "Anodized surface designs presented were taken with a scanning electron microscope pictures with different magnification fragments anodized surfaces of test samples: with biocorrosion damage, chemical corrosion and control samples with exposure time of 40 days (table 1), 120 days (table 5) and 180 days (table 9).

In tables 2, 6, 8 "Chemically polished surface of the samples" presents pictures with different magnification fragments chemically polished surfaces of test samples: with biocorrosion damage, chemical corrosion and control samples with exposure time of 40 days (table 2), 120 days (table 6) and 180 days (table 8).

In tables 3, 4, 7 the Surface of the samples after mechanical treatment (exposure to 40 days)" presents pictures with different magnification fragments surfaces and the testing of the samples after mechanical processing: biocorrosion damage, chemical corrosion and control samples with exposure time of 40 days (table 3), 120 days (table 4) and 180 days (table 7).

As a result of the test shows that on the surface of the aluminum-magnesium alloy as the result of biological corrosion is undergoing profound changes of the surface increases with the duration of exposure of the consortium of microorganisms.

Evaluation of bio-corrosion specimens: the type definition of bio-corrosion, and the distribution of corrosion damage on the surface of the samples, carried out according to GOST 9.908-85.

So, on the anodized surface of the test specimen visible spots oval, cracks, large and small hollows (table 1); visible deep cracks and flaking crust (table 5); spots, loose cortex (table 9); the surface of the control sample: covered with small pores uniformly distributed throughout the surface of the sample (table 1), became scaly (table 5), covered with a network of cracks (table 9).

Chemically polished surface of the test specimen visible: thin dehiscent, partially flaking crust and a small ulceration oval (table 2); thick dehiscent cork and traces of mycelium consortium (table 6); elongated and sticky flakes (table 8); in the control sample are visible: surface ijazul the deposits (table 2), large and small hollows (table 6) and rounded flat spots (table 8).

On the surface of the test specimen after machining visible subsurface ulcers rounded (table 3), was formed dehiscent cortex, sometimes resulting in grainy, traces of the original surface topography visible with difficulty (table 4), on the surface visible rounded scales (table 7); in the control sample visible defects in the form of furrows (table 3), significant disturbance of surface topography (table 4), the surface is covered with dehiscent cortex (table 7).

In the study of test samples of aluminum-magnesium alloy subjected to longer trials (180 days) on biological corrosion, revealed noticeable changes in the surface compared to the control samples.

When comparing the appearance of metal samples that were exposed to bio-corrosion, it is noticeable that the greatest changes were surface samples after mechanical processing.

Comparison destruction of biological and chemical corrosion led to the conclusion that these processes are different in outward appearance. Chemical corrosion does not cause profound changes of aluminum-magnesium alloy. The processes of bio-corrosion have their own characteristics, namely, the increased heterogeneity of impacts on the surface Metalli his deep subsurface defeat of metabolic products of fungi. Corrosion distribution of lesions on the surface of the test sample is uneven.

Thus, biocorrosion damage are complex and cannot be attributed to any one type of corrosion, and are a combination of several types of corrosion: localized corrosion and corrosion spots, characterized by small corrosion lesions of irregular shape (primarily anodized surface of the test samples), intergranular corrosion, characterized by the presence of corroded zone along the grain boundaries of the metal, affecting the boundaries of all grains or grains (primarily chemically polished surface of the test samples), subsurface corrosion, characterized by the fact that corrosion damage is on the surface of small area and mainly focused under the surface of the metal (primarily the surface of the test pieces after machining).

1. Method qualitative assessment biocorrosion lesions thin-walled hermetic shell of aluminium-magnesium alloys with the operation of spacecraft, characterized by the fact that made the test and control samples of aluminum-magnesium SPLAVE is, prepared samples are dried, sterilized, for growing culture of the fungi spores inoculated in tubes with beveled agar medium of čapek-doksa, thermostatic tubes at a temperature (29±2)°C for 14-28 days before the appearance of Mature spores, preparing a suspension of spore material, which is applied to sterilized test samples, then dried them and put one sample in Petri dishes on agar surface environment of čapek-doksa, control samples not treated spore material, also feature on the surface of the agar medium of čapek-doksa in Petri dishes, Petri dishes with test and control samples are placed in different executory, the bottom of which pour water to maintain humidity over 90%, executory closed and incubated at a temperature (29±2)°C, conducting exposure of the test and control samples, then the samples are then removed from the Petri dishes, remove the corrosion products and the mycelium from the sample surface, rinsing them under running water, identify indicators biocorrosion lesions, characterized in that environment čapek-doksa prepare a spore suspension of materials from individual cultures of fungi - bacteria strains of Paecilomyces variotii Bainier BKM F-4039D, Ulocladium botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thorn BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Churh BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D, which are then mixed in equal proportions, and the concentration of spores of each species of fungus in the suspension should be in the range of 1-2 million/cm3obtained suspension by means of a spray is applied on the test samples of aluminum-magnesium alloys with different kinds of surface treatment - machined, chemically polished, anodized, after washing the samples in flowing water, they kept in 70%ethyl alcohol for 30 min, then washed with detergent and dried, then using a scanning electron microscope conduct a qualitative assessment biocorrosion lesions: change the appearance of the surface of the sample estimate of the initial stages of bio-corrosion and its type, the distribution of corrosion damage on the sample surface, and determine the dependence biocorrosion process from the time when the exposure test and control samples performed within 40, 120 and 180 days.

2. The spore suspension material including a mixture of spore-bearing materials from individual cultures of fungi Paecilomyces variotii Bainier BKM F-4039D, Ulocladiurn botrytis Preuss BKM F-4032D, Penicillium chrysogenum Thorn BKM F-4034D, Aspergillus sydowii (Bainier et Sartory) Thom et Church BKM F-4037D, Cladosporium sphaerospermum Penz. BKM F-4041D in equal proportions, to assess biocorrosion lesions thin-walled hermetic shell of the brand is ievo-magnesium alloys with the operation of spacecraft.



 

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