Научная статья на тему 'The structure and properties of microbiocenosis in dumps of the fuel and energy complex of Ukraine'

The structure and properties of microbiocenosis in dumps of the fuel and energy complex of Ukraine Текст научной статьи по специальности «Биологические науки»

CC BY
76
13
i Надоели баннеры? Вы всегда можете отключить рекламу.
Журнал
Biotechnologia Acta
CAS
Область наук
Ключевые слова
C RYPTOCOCCUS ALBIDUS / -L-RHAMNOSIDASE / NARINGIN / NEOHESPERIDIN / RUTIN / FLAVONOIDS / CITRUS JUICES / GREEN TEA / ABORIGINAL COMMUNITY / DUMPS / BIOLEACHING / АБОРИГЕННЕ УГРУПОВАННЯ / ВІДВАЛИ / БіОВИЛУГОВУВАННЯ / АБОРИГЕННОЕ СООБЩЕСТВО / ОТ ВАЛЫ / БИОВЫЩЕЛАЧИВАНИЕ

Аннотация научной статьи по биологическим наукам, автор научной работы — Blayda I.A., Vasyleva T.V., Sliusarenko L.I., Shuliakova S.N., Хитрич В. Ф.

The work aimed to conduct complex chemical and microbiological study of the dumps of the fuel and energy complex of Ukraine. It is established that the qualitative composition of the aboriginal microbiota of the studied technogenic substrates does not to depended on the storage time, because it was determined by the chemical and mineralogical compositions and is mainly represented by the heterotrophic and acidophilic chemolithotrophic bacteria (AСB). It is noted that the number of all groups of microorganisms in dumps increased during long term storage due to internal processes and the impact of external climatic factors. In our experiment the ACB association demonstrated the maximum leaching activity when the divalent iron was as an energy source. It is also noted that the “silicate” bacteria present in the aboriginal consortium and have no leaching activity, significantly increase bioleaching rates by ACB. The results of the study indicate on the formation of resistant specific microbiocenoses in the dumps of the fuel and energy complex that can be used as sources of highly active strains obtaining for use in biotechnological processes of metal extraction.

i Надоели баннеры? Вы всегда можете отключить рекламу.
iНе можете найти то, что вам нужно? Попробуйте сервис подбора литературы.
i Надоели баннеры? Вы всегда можете отключить рекламу.

Текст научной работы на тему «The structure and properties of microbiocenosis in dumps of the fuel and energy complex of Ukraine»

UDC 579:662.7 https://doi.org/10.15407/biotech11.05.054

THE STRUCTURE AND PROPERTIES OF MICROBIOCENOSIS IN DUMPS OF THE FUEL AND ENERGY COMPLEX OF UKRAINE

I. A. Blayda T. V. Vasyleva

L. I. Sliusarenko Odesa National Mechnykov University, Ukraine

S. N. Shuliakova V. F. Rhitrich

E-mail: [email protected]

Received 19.05.2018 Revised 09.08.2018 Accepted 30.10.2018

The work aimed to conduct complex chemical and microbiological study of the dumps of the fuel and energy complex of Ukraine. It is established that the qualitative composition of the aboriginal microbiota of the studied technogenic substrates does not to depended on the storage time, because it was determined by the chemical and mineralogical compositions and is mainly represented by the heterotrophic and acidophilic chemolithotrophic bacteria (ACB). It is noted that the number of all groups of microorganisms in dumps increased during long term storage due to internal processes and the impact of external climatic factors. In our experiment the ACB association demonstrated the maximum leaching activity when the divalent iron was as an energy source. It is also noted that the "silicate" bacteria present in the aboriginal consortium and have no leaching activity, significantly increase bioleaching rates by ACB. The results of the study indicate on the formation of resistant specific microbiocenoses in the dumps of the fuel and energy complex that can be used as sources of highly active strains obtaining for use in biotechnological processes of metal extraction.

Key words: aboriginal community, dumps, bioleaching.

The fuel and energy complex (FEC) of Ukraine produces waste products as a result of the coal mining and processing. Accumulating in huge quantities at the territories of industrial complexes, the waste creates an additional burden on the environment. At the same time, the waste of FEC contains, in particular, rare metals in industrial concentrations, which makes it a "man-made deposit", the unconventional raw materials of valuable components [1, 2]. A special aboriginal microbial community is formed under the influence of industrial and natural factors, and later affected by the storage conditions in the studied anthropogenic ecosystems. In long-term storage, the substrates change affecting the structure and composition of the microbiocenosis and the ability of the formed equilibrium systems to destruct. The available literature data suggests that the use of the aboriginal consortium associations in the leaching metals biotechnology is promising due to the effect of syntrophic relationships between individual groups of microorganisms in the

community [3, 4]. The microbial biotechnologies should be implemented based on comprehensive studies of the biological and physicochemical properties of the initial solid substrate, the qualitative and quantitative assessment of the indigenous microbial community present in it, the possibility of isolating, selecting and selecting the most promising highly active strains. There is ample evidence of the microbial role in both the formation and destruction processes of geogenic substrates (natural ores and minerals, native sulfur, oil, peat, coal, etc.), accompanied by the bioextraction of useful components [5-7]. Information is limited about the life and biochemical activity of microorganisms in the raw materials of technogenic origin.

The aim of the work was to conduct a comprehensive chemical and biological research of technogenic raw materials produced by the FEC enterprises of Ukraine to establish the structure and properties of their microbiocenosis.

Material and Methods

The objects of research were dumps of the Central concentrating plant (CCP) of the Lviv-Volyn coal basin (LVCB), fly ash and ash from the burning of LVCB coal at Ladyzhinska and Dobrotvorska TPSs, respectively. The samples (more than 60 in total) were taken during 2008-2014 (April to November) on the slopes of the dumps at the 50.0 ± 5.0 cm surface layer.

To identify various physiological groups of native microbial microorganisms, enrichment cultures and specific nutrient media were used: 9K and 9K * for the acidophilic chemolithotrophic bacteria (ACB); Beyerinck for the neutrophilic chemolithotrophic bacteria; 882 for representatives of the genus Leptospirillum; 150a for the moderately thermophilic representatives of the genus Acidithiobacillus, such as A. caldus; Gorbenko for the heterotrophic bacteria; Czapek for the filamentous fungi; and A-27 for the "silicate" bacteria (Table 1) [8, 9].

As the energy source, either FeSO47H2O at a concentration of 44.5 g/dm3, or elemental sulfur or thiosulfate at a concentration of

5.0 g/dm3 was added to the mineral background of the 9K medium.

The biomass of various representatives of the dump microbiocenosis was accumulated at a ratio of solid (substrate) to liquid (nutrient medium) S: L = 1: 10. In the control experiments sterile substrate was introduced to the nutrient medium. The cultures were incubated at a temperature of 30.0 ± 0.5 °C for mesophilic (MP) and 50.0 ± 0.5 °C for moderately thermophilic (MTP) bacteria, pH 3.0-7.0 for 5 days. The development of microorganisms was evaluated by the presence of the surface film, the change in pH and the appearance of the bacterial suspension. The abundance of representatives of different microbial groups was established by sowing tenfold serial dilutions of the bacterial suspension on agar media of the same composition. The number of spore-forming bacteria was determined after heat treatment at 80.0 ± 0.5 °C for 15 minutes.

The biogeochemical activity of the aboriginal community was judged by the concentration of metals transferred from the solid phase to the culture medium. Selective nutrient media were used as leaching solutions

Table 1. The composition of nutrient media (g/dm3) for identification of microbial groups in waste products of the fuel and energy complex of Ukraine

Mineral components Culture media [8, 9]

A-27 Czapek 9K 9K* 150a Beyerinck 882

KH2PO4 0.50 0.05 0.50 0.027

(NH4)2SO4 3.00 0.45 3.00 0.132

MgSO4-7H2O 0.50 0.50 0.50 0.50 0.50

MgCl2-6H2O 0.10 0.053

nh4cl 0.10

KCl 0.50 0.10 0.05 0.10

NaNO3 3.00

K2HPO 1.00

Na2HPO4 2.00 0.20

Ca(NO3)2 0.01 0.014 0.01

CaCl2-2H2O 0.147

FeCl3 0.001 0.01

NaHCO3 1.00

CaCO3 1.00

Quartz 10.00

Sucrose 5.00 30.00

Yeast extract 0.02

pH 7.5-8.0 6.6-7.2 1.0-3.5 1.8-4.0 1.5-3.5 4.5-8.5 2.0-4.0

(Table 1). Sterile waste with a sterile leach solution served as controls. The bioleaching process was carried out by the vat method at a ratio S: L = 1:10, pH — 2.0, 30.0 ± 0.5 °C for MP and 50.0 ± 0.5 °C for MTP bacteria for 7 days. The concentration of metals in solid substrates and solutions was determined by atomic absorption spectroscopy on AAS-1 (Germany) and C-115PK Selmi (Ukraine) devices [10]. The reliability of obtained results was evaluated by the Student's i-test with a probability of P < 0.05.

Results and Discussion

Table 2 shows the chemical composition of the studied substrates by the main components.

To observe the development of aboriginal associations in substrates, depending on the timing of their accumulation, preliminary microbiological crops were grown on nutrient media selective for acidophilic chemolithotrophic and heterotrophic microorganisms because those are typical representatives of microbiocenosis of geogenic and technogenic origin (Table 3).

The microorganisms were not detected at all or their communities were very poor in fresh substrates, especially heat-treated ash and fly ash. In storage, the communities are formed. In dumps with an acidic environment, the conditions favor the active growth of both chemolithotrophic and heterotrophic microorganisms. In neutral or weakly alkaline ash and fly ash, the heterotrophic component forms an association faster than the chemolithotrophic one, and quantitatively the

former is more pronounced, both in comparison with the dumps and in relation to its own chemolithotrophic component. However, generally, stable, numerous aboriginal equilibrium communities form during the storage of the studied technogenic substrates for longer than three years. Then it is possible to isolate active strains for biotechnological developments. For further research, ash and fly ash (storage period 24-36 months), as well as "stale" dumps substrates with different shelf life were chosen: 24-28 months (black) and more than 60 months (red).

The results of microbiological studies are shown in Fig. 1. They indicate the quantitative prevalence of heterotrophic and acidophilic chemolithotrophic bacteria, both mesophilic and moderately thermophilic, in waste products of FEC.

In all the studied substrates, especially with acidic pH (black and red coal preparation waste), ACB dominate which use bivalent iron and sulfur / thiosulfate as an energy source. In the mesophilic association of all substrates that develops on a standard 9K medium with bivalent iron or thiosulfate, a lot of small Gram-negative rod-shaped cells were noted. In the microbial associations of iron and sulfur-oxidizing bacteria, no significant differences in cell morphology were seen in accordance with the available literature data. The number of cells oxidizing bivalent iron and thiosulfate in ferrous dumps, fly ash and ash was 104-105 cells/g.

In the red "stale" dumps, their number was greater and reached 108 and 106 cells/g for iron-

Table 2. The content of main controlled metals in waste products of FEC of Ukraine (g/ton)

Metal Minimum industrial content Ash, Dobrotvorska TPS Fly ash, Ladyzhinska TPS Dumps of coal concentrating, CCP "Chervonohradska"

Black Red

Plumbum 18-22 75.0±0.05 120.0±0.1 42.20±0.05 57.92±0.05

Nickel 80-120 110.0±0.1 170.0±0.1 134.2±0.1 132.9±0.1

Cadmium 45,0-55,0 8.5±0.05 7.5±0.05 2.82±0.05 3.63±0.05

Aluminum (2.5-5.0)103 (105.0±0.1)103 (37.5±0.05)103 (13.92±0.05)103 (8.92±0,05)103

Cuprum 80-100 92.5±0.1 60.0±0.05 62.18±0.05 78.90±0.05

Manganese 850-1000 1750±0.1 600.0±0.1 317.7±0.1 812.9±0.1

Zinc 65-70 110-180 315.0±0.1 112.5±0.1 130.9±0.1

Germanium 15-20 30-40 45.0±0.05 26.0±0.1 30.0±0.1

Gallium 15-20 30-40 95.0±0.1 15.1±0.1 22.4±0.1

Table 3. Microbial characteristic of waste products of FEC of Ukraine

Substrate Storage time рН of water extract Microbial abundance, cell/g

Heterotrophic Acidophilic chemolithotrophic

Bacteria Fungi Oxidizing Fe(II) Oxidizing S0

Dumps Freshly produced 1.8-2.0 100±5 50±2 (5.3±0.4)x102 15±5

Dumps 10-14 months 2.0-2.4 (3.60±0.65)x103 (1.50±0.25)x102 (4.70±0.85)x103 (1.55±0.25)x102

Dumps 24-28 months 2.6-3.3 (7.80±1.65)x105 (8.70±0.75)x102 (7.50±1.56)x105 (7.70±0.55)x104

Dumps > 36 months 3.0-3.6 (5.60±1.25)x107 (6.70±1.35)x103 (9.35±1.85)x107 (4.80±0.95)x105

Dumps > 60 months 3.5-4.5 (9.30±1.85)x107 (1.50±0.30)x104 (3.70±0.75)x108 (3.95±0.75)x106

Ash Freshly produced 5.8-6.2 15±3 n/o* n/o n/o

Ash 10-14 months 6.0-7.0 (8.20±1.65)x103 (2.70±0.55)x102 (5.80±1.15)x102 (3.50±0.70)x102

Ash 20-24 months 6.2-7.4 (7.95±1.58)x105 (8.75±1.75)x102 (2.80±0.55)x104 (4.30±0.85)x103

Ash 24-36 months 6.4-7.8 (4.70±0.96)x107 (2.10±0.45)x103 (3.55±0.75)x105 (4.55±0.95)x104

Fly ash Freshly produced 6.4-8.0 9±1 n/o n/o n/o

Fly ash 10-14 months 7.0-9.5 (5.20±1.1)x103 (4.30±0.85)x102 (9.70±1.95)x102 (4.15±0.80)x102

Fly ash 20-24 months 7.8-10.2 (8.55±1.75)x105 (2.75±0.55)x103 (1.25±0.25)x103 (9.70±1.95)x102

Fly ash 24-36 months 9.0-10.8 (2.55±0.55)x108 (7.70±1.55)x103 (8.70±1.75)x104 (1.40±0.25)x103

* — not observed.

and sulfur-oxidizing bacteria, respectively. The obtained data suggest the presence of representatives of the genus Acidithiobacillus (A. ferrooxidans and A. thiooxidans), widely distributed in natural sulfide ores in the studied technogenic raw material and actively participating in metal leaching processes [11].

In mesophilic conditions of medium 882, morphologically different cells — spirillus, vibrios, and small curved rods — were observed. Their number in black waste, fly ash, and ash was 103-104 cell/g, in the red "stale" dumps it was 105 cell/g. This suggested the presence of representatives of the genus Leptospirillum in the studied microbiocenosis. The genus includes iron chemolithotrophic bacteria [12]. These results are consistent with the available literature data, according to which bacteria of the genus Leptospirillum are always present in natural and man-made mineral raw materials and do not make a significant contribution to the bioleaching processes themselves, but in an aboriginous consortium with A. ferrooxidans and A. thiooxidans they contribute to the efficiency of extraction metals [13].

In mesophilic conditions of Beyerinck's medium, the development of small Gramnegative rods was observed in the amount of 102 cell/g (black dumps, fly ash and ash) and

104 cell/g in red "stale" dumps. This suggested the presence of neutrophilic thionic bacteria of the genus Thiobacillus in the studied microbiocenoses. There is no data on the presence of this group of bacteria in technogenic raw materials, as well as any information about their ability to leach metals. However, it is known that in nature, Thiobacillus thioparus in association with Thiobacillus ferrooxidans are active agents of corrosion of metallic and non-metallic products. Therefore their presence in biocenoses with quantitative advantage of acidophilic bacteria is quite possible [14].

The microbiocenosis of FEC waste products is formed in widely ranging temperatures during storage. There is also the self-heating and self-ignition phenomena. Thus it was interesting to identify bacteria for which the optimum temperature for growth is 45.0-50.0 °C. An insignificant amount (103-104 cell/g) of small gram-negative rod-like cells was noted in all substrates in the cumulative culture based on the medium 150a. This can be attributed to the genus Acidithiobacillus, in particular A. caldus. According to a number of studies, A. caldus is always, albeit slightly, present in mineral raw materials and, together with A. ferrooxidans and A. thiooxidans, contributes to the efficiency of metal extraction [15].

MIT ACE. oaxliiing Feçn) 30.0% \

SporogHitui 7.0%

NoïL-sfKOfigeûoys 13.0%

MTPACB. i-ivi.Hi-mp s13 20.0 %

Hl SiLcate" bacteria 1,0 %

\ MP ACE. Willing S*1 3 0 H

MP ACE, pe(II) 26,0 %

a

MP ACE. oxidizing FcQI) 35,0%

MTP ACE. ftKidinDg 47,0%

kfPACB.MtidlELilg 2.0%

"Silicate" bacteria 2.0 % , Non-iporogenouj 6,0 H

.y f".r i.: 2,0 %

MTPACB, tundra^«® 6,0*

MPACE 15,D%

MTP ACE 21,0%

Het« oil 0|>liic 95.0°«

iНе можете найти то, что вам нужно? Попробуйте сервис подбора литературы.

Heterotrophic 64

MTPACB.1,0"

Fig. 1. Quantitative and qualitative content of microbial cenoses: a — black dumps of CCP; b — red dumps of CCP; c — fly ash of Ladyzhinska TPS; d — ash of Dobrotvorska TPS

Mixotrophic, moderately thermophilic bacteria were found in all studied waste substrates. In a cumulative culture based on a modified 9K medium with thiosulfate or bivalent iron, an abundant development of Gram-positive short round or cocco-like spore-forming cells was recorded, which can be attributed to the genus Sulfobacillus. Their numbers were much higher in red "old" waste and reached almost 109 cell/g. This is the maximum quantitative indicator among all the iron- and sulfur-oxidizing bacteria identified in the studied substrates, possibly associated with the processes leading to self-heating of the dumps.

The FEC waste products are bio-inert systems. However, an intensive development of heterotrophic bacteria, both spore-and non-spore-forming (Fig. 1) was observed there, with numerical predominance in ash and fly ash with neutral and weakly alkaline pH (up

to 108 cell/g), as well as in the "stale" coal enrichment dumps, up to 107 cell/g. These results are consistent with available reports on the ability of certain representatives of heterotrophic bacteria to grow in mineral solutions in the presence of trace amounts (pg/dm3) of organic substrates, as well as the activity of the representatives of the genera Pseudomonas and Bacillus in the leaching of gold and uranium [16].

The presence of so-called "silicate" bacteria capable of destroying silica and silicates was first established in the heterotrophic component of FEC waste products. In bacterial suspension on nutrient A-27 medium, Grampositive large rods capable of forming spores were recorded, on agarized A-27 medium they were formed in almost identical round transparent colorless colonies. Their number was in the range of 103-104 cell/g, reaching a maximum in ash (105 cell/g), which may be

b

d

c

due to the increased content of silicon and aluminum in this substrate (Table 1) [17].

Thus, the qualitative composition of the microbiocenoses of the FEC waste products under study does not depend on the storage time, since it is determined by the chemical and mineralogical composition of the substrates. As they accumulate and are stored under the influence of external factors, quantitative differences arise in the microbiocenoses, which are affected by the accumulation time, composition and pH of waste products. When comparing substrates with the same storage time but different pH, it is obvious that in ash (pH 6.4-7.8) and fly ash (pH 9.0-10.8) the conditions are more favorable for the development of heterotrophic microorganisms both in comparison with the black dumps (pH 2.6-3.3), and in relation to the development of own acidophilic chemolithotrophic component (Fig. 1). It should be noted that the abundance of the ACB community in fly ash (amorphous, finely dispersed, with a large specific surface), despite its alkaline pH, is comparable to the coarse ash. This is another important factor for bioleaching associated with the presence of microorganism cells, either free or attached to the surface of solid particles. The presence of defects in the crystal structure and a large specific surface of the substrate contribute to faster growth and development of all possible microorganisms of the consortium [18].

The biotechnological potential of representatives of the consortium of FEC waste products was determined by their ability to create favorable conditions for certain groups of microorganisms to destroy substrates and extract valuable metals from those.

Representatives of acidophilic chemolitho-trophic bacteria oxidizing bivalent iron and thiosulfate were the most numerous group of practical interest in the aboriginal consortium of FEC waste products. The results on leaching of metals from FEC waste products by the most numerous groups in the aboriginal consortium, the mesophilic and moderately thermophilic association of ACB, are presented in Fig. 2 and 3 respectively. In the control experiments with sterile substrates and nutrient mediums the leaching of Ge, Ga, Cd, Ni, Cu, Zn, Mn and Al did not exceed 2-4%, the leaching of Pb did not exceed 0,3-0,5%.

Hence, the association of moderately thermophilic bacteria of the studied substrates, regardless of their nature, was distinguished by a higher leaching activity compared to the mesophilic community. However, maximum

leaching rates of metals, both rare and heavy, were achieved using bivalent iron as an energy source similarly in mesophilic and moderately thermophilic conditions. This confirms the leading role of A. ferrooxidans in the processes of bacterial leaching metals under mesophilic conditions [9, 18, 19]. It is established that the black coal dumps is the most "accessible" to microorganisms; the degree of leaching of almost all registered metals exceeds the similar indicators for other substrates.

The change in the microbial landscape and the number of bacteria was studied during the entire experiment of metal bioleaching of from the black dumps (Fig. 4). Thus, during the first day, the number of bacteria in the mesophilic association did not exceed 4.5x102 cell/g. Gram-negative short thin cells prevailed in the stained microscopic preparation, sometimes larger Gram-positive cells with rounded ends were encountered (Fig. 4, a). After five days, the number of bacteria increased significantly and reached 7.8x107 cell/g. During this period of time, bloated round cells with thickened membrane and unstained contents were recorded in the bacterial suspension (Fig. 4, b). Bipolar inclusions were present on the surface of some cells. According to available literature data, those are globules of sulfur resulting from the oxidation of mineral raw materials [20].

The appearance of rounded large cells was also observed during longer-term cultivation of the studied strains (Fig. 4, c). After 7 days their total amount in the bacterial suspension decreased to 5.7x 104 cell/g. This was accompanied by lysis of the cells, a shift in the pH of the leach solution towards neutral values and the beginning of deposition of insoluble Fe+3 compounds.

When using the moderately thermophilic association of ACB, the number of bacteria was slightly higher. At the beginning of the experiment it was 5.7x104 cell/g, within five days it reached a maximum of 7.3x1010 cell/g and at the end of the experiment (after 7 days) decreased to 3.9x105 cell/g. This changing pattern of biomass amount is a reflection of the classical phases of growth and development of microorganisms: exponential, stationary, dying off and cell lysis phases [9].

Despite the quantitative advantage of the heterotrophic component in the waste product microbiocenosis (Fig. 1), its leaching activity was insignificant and the extraction of metals into the solution did not exceed 15.0%. Extraction of metals into the solution as a result of the activity of "silicate" bacteria was also

tf*-

OÔ c

s

a

a

90

80

S 70

r. 60

50

40 30 20 10

0

*

8

i

mm

in II I

s nnam i

i iinii

*

[

Ge Ga Cd | Hi J Cu Pb Za Mh A1 Black dumps

Ge Ga Cd Ni Cu Pb Zn Mn A1 Red dumps

■ Iron aThio sulfate

! II «11

i

Ge Ga Cd Ni Cu Pb Zn Mn A1 Fly ash Ash

■ Iron ■Thiosulfate

Fig. 2. Metal leaching by the mesophilic association of acidophilic chemolithotrophic bacteria from the FEC

waste products

Hereinafter: *P < 0.05 compared with control In the control experiments with sterile substrates and nutrient mediums the leaching of Ge, Ga, Cd, Ni, Cu, Zn, Mn and Al did not exceed 2-4%, the leaching of Pb did not exceed 0,3-0,5%

low (it did not exceed 12%). However, it was obvious that the substrates were destroyed and their appearance changed after contacting with the nutrient medium A-27, creating favorable conditions for the growth and activity of "silicate" bacterial consortium (Fig. 5).

The waste products contained a lot of silicon-containing phases (silica, aluminosilicates). Hence our assumption that the association of "silicate" bacteria at the

initial stage of processing the FEC dumps due to the destruction of stable crystalline silicate structures can enhance the effectiveness of the further action of the chemolithotrophic component of the consortium in relation to metal recovery. This idea was confirmed (Fig. 6), the results are patented [21].

Thus, the complex chemical and microbiological studies of waste products of FEC of Ukraine showed that the physico-

Fig. 3. Metal leaching by the moderately thermophilic association of acidophilic chemolithotrophic bacteria from the FEC waste products

b

Fig. 4. Micrographs of the association of mesophilic ACB in the metal leaching from the black dumps:

at the beginning of the process (a), after 5 (b) and 7 (c) days; x1000

a

c

Fig. 5. Appearance of black waste dumps:

before (a) and after (b) bacterial leaching with nutrient medium A-27

Fig. 6. Extraction of germanium and gallium from the waste dumps:

by the mesophilic association of acidophilic chemolithotrophic bacteria (a), after pre-treatment with medium

A-27 (b). Leaching time 24 hours

chemical composition and the conditions of their formation and storage determine the structure of their microbiocenosis. The qualitative composition of the native microbiota does not depend on the storage time. The microbiota is determined by the chemical and mineralogical composition of substrate, and is represented mainly by heterotrophic and acidophilic chemolithotrophic bacteria. An increase in the number of all groups of microorganisms in long-term stored waste products is established, which is a result of internal processes and the influence of external climatic factors. The

maximum leaching activity is observed in the ACB associations, especially when using bivalent iron as an energy source. The "silicate" bacteria found in the structure of the aboriginal consortium are not capable of leaching metals on their own. However in combination with ACB they contribute to a significant increase in the bioleaching rates. The research results indicate the formation of stable specific microbiocenoses in FEC waste products. The coenoses can be sources of obtaining highly active strains for use in biotechnological processes of metal extraction.

REFERENCES

1. Galetskii L. S., Naumenko U. Z., Pilipchik A. D. Technogenic deposits are a new non-traditional source of mineral raw materials in Ukraine. Ekolohiia dovkillia ta zabezpechennia zhyttediialnosti. 2002, 5(6), 77-81. (In Ukrainian).

2. Pashkov G. L., Saykova S. V., Kuz'min V. I. Ash of natural coals is an unconventional source of raw materials of rare elements. Zhurnal Sibirskogo federalnogo universiteta. Seriia: Tekhnika i tekhnologii. 2012, V. 6, P. 520530. (In Russian).

3. Blayda I. A. Extraction of valuable metals during industrial wastes processing by biotechnological methods (Review). Energo-tehnologii i resursosberezhenie. 2010, V. 6, P. 39-45. (In Russian).

4. Blaida I. A., Vasileva T. V., Sliusarenko L. I., Khitrich V. F., Ivanytsia V. A. Extraction of rare and nonferrous metals by microbial communities of the ash from burning Pavlograd's coal. Mikrobiologiya i Biotekhno-logiya. 2012,V. 3, P. 91-101. (In Russian).

5. Karavayko G. I., Kuznetsov S.I., Golomzik A.I. The role of microorganisms in leaching metals from ores. Moskva: Nauka. 1972, 248 p. (In Russian).

6. Ivanov M. V., Karavayko G.I . Geological microbiology. Mikrobiologiya. 2004, 73(5), P. 581-597. (In Russian).

7. Tolstov E. A., Latyshev V. E., Lil'bok L. A. Possibilities of using biogeotechnology in leaching of poor and refractory ore. Gornyj zhurnal. 2003, V. 8, P. 63-65. (In Russian).

8. Methods for General Bacteriology. V. 2. Moskva: Mir. 1984, 265 p. (In Russian).

9. Karavayko G. I., Rossi Dzh., Agate A. Biotechnology of metals. A Practical Guide. Moskva: Tsentr mezhdunarodnykh proektov GKNT. 1989, 375 p. (In Russian).

10. Khavezov I., Tsalev D. Atomic Absorption Analysis. Leningrad: Khimiya. 1983, 144 p. (In Russian).

11. Vasileva T. V., Blaida I. A., Ivanytsia V. A. The main groups of microorganisms involved in the biohydrometallurgical process. Problemy ekolohichnoi biotekhnolohii. Available at http://jrnl.nau.edu.ua/index.php/ ecobiotech/ article/view/4678 (accessed, June, 2013).

12. Shuang Mi, Jian Song, Jianqun Lin, Yuanyuan Che, Huajun Zheng, Jianqiang

Lin. Complete Genome of Leptospirillum ferriphilum ML-04 Provides Insight into Its Physiology and Environmental Adaptation. The Microbiological Society of Korea. 2011, 49(6), 890-901. https://dpi.org/10.1007/ s12275-011-1099-9

13. Giaveno A., Lavalle L., Chiacchiarini P., Donati E. Bioleaching of zinc from low-grade complex sulfide ores in an airlift by isolated Leptospirillum ferrooxidans. Hydrometallurgy. 2007, 89(1-2), 117-126. https://dpi.org/10.1016/j.hydromet. 2007.07.002

14. Andreyuk E.I., Kozlova I.A. Lithotrophic bacteria and microbiological corrosion. Kiyv: Naukova dumka. 1977, 164 p. (In Russian).

15. Zhou Qiu Guan, Bo Fu, Hong Bo Zhou. Isolation of a strain ofAcidithiobacillus caldus and its role in bioleaching of chalcopyrite. World J. Microbiol. Biotechnol. 2007, 23(9), 1217-1225.

16. Kukanova S. I. Heterotrophic microorganisms and their role 3 processes of gold extraction from non-standard raw materials. Ph.D. dissertation. Dept. Rudnoi mikrobiologii i bio-geotehnologii. Institut mikrobiologii Respubliki Uzbekistan, Tashkent, 1992. (In Russian).

17. Karavayko G. I., Belkanova N. P., Eroshchev-Shak V. A., Avakyan Z. A. The role of microorganisms and some physicochemical factors of the environment in the destruction of quartz. Mikrobiologiya. 1984. V. 53(6), P. 976-981. (In Russian).

18. Torma A. E. The role of Thiobacillus ferro-oxidans in hydrometallurgical processes. Adv. Biochem. Engin. 1977, 6, 1-37. https:// dpi.org/10.1007/3-540-08363-4_1

19. Tributsch H. Direct vs indirect bioleaching. Hydrometallurgy. 2001, 59(2-3), 177185. https://dpi.org/10.1016/S0304-386X(00)00181-X

20. Sokolova G. A., Karavayko G.I. Physiological and geological activity of thiobacteria. Moskva: Nauka. 1964, 332 p. (In Russian).

21. Blaida I. A., Vasileva T. V., Semenov K. I., Baranov V. I., Ivanytsia V. A. A two-stage method of bioleaching of gallium and germanium. UA Patent 104268. January 25, 2016. (In Ukrainian).

СТРУКТУРА ТА ВЛАСТИВОСТ1 М1КРОБ1ОЦЕНОЗ1В В1ДХОД1В ПАЛИВНО-ЕНЕРГЕТИЧНОГО КОМПЛЕКСУ УКРА1НИ

I. А. Блайда Т. В. Васильева Л. I. Слюсаренко С. М. Шулякова В. Ф. Хитрич

Одеський нащональний ушверситет iMern I. I. Мечникова, Укра!на

E-mail: [email protected]

Метою роботи було проведення комплексного хiмiко-мiкробiологiчного досл^ження в^вальних продуктiв паливно-енергетич-ного комплексу Укра!ни. Встановлено, що якiсний склад аборигенно! мiкробiоти досль джених техногенних субстратiв не залежить вщ термiнiв зберiгання, оскiльки визнача-еться хiмiчним i мiнералогiчним складом, i представлений переважно гетеротрофними i ацидофiльними хемолiтотрофними бактерь ями (АХБ). Виявлено зростання чисельност всiх груп мiкроорганiзмiв у вiдвальних продуктах тривалого збер^ання, що е результатом внутр^шх процесiв i впливу зовшшшх клiматичних факторiв. Показано, що мак-симальну вилуговувальну активнiсть мають асоцiацii АХБ, особливо в разi використання двовалентного залiза як джерела енергп. Ви-явленi в структурi аборигенного консорцiуму «сил^атш» бактерii, що не здатш вилугову-вати метали самосийно, в поеднаннi з АХБ сприяють значному пiдвищенню показникiв бшвилуговування. Результати дослiджень свiдчать про формування у в^вальних продуктах паливно-енергетичного комплексу стiйких специфiчних мiкробiоценозiв, що можуть бути джерелами отримання високо-активних штамiв для використання в б^тех-нологiчних процесах вилучення металiв.

Ключовi слова: аборигенне угруповання, вщвали, бiовилуговування.

СТРУКТУРА И СВОЙСТВА МИКРОБИОЦЕНОЗОВ ОТХОДОВ ТОПЛИВНО-ЭНЕРГЕТИЧЕСКОГО КОМПЛЕКСА УКРАИНЫ

И. А. Блайда Т. В. Васильева Л. И. Слюсаренко С. Н. Шулякова В. Ф. Хитрич

Одесский национальный университет имени И. И. Мечникова, Украина

iНе можете найти то, что вам нужно? Попробуйте сервис подбора литературы.

E-mail: [email protected]

Целью работы было проведение комплексного химико-микробиологического исследования отвальных продуктов топливно-энергетического комплекса Украины. Установлено, что качественный состав аборигенной микро-биоты исследованных техногенных субстратов не зависит от сроков хранения, поскольку определяется химическим и минералогическим составом, и представлен в основном гетеротрофными и ацидофильными хемоли-тотрофными бактериями (АХБ). Выявлено возрастание численности всех групп микроорганизмов в отвальных продуктах длительного хранения, что является результатом внутренних процессов и воздействия внешних климатических факторов. Показано, что максимальной выщелачивающей активностью обладают ассоциации АХБ, особенно при использовании двухвалентного железа в качестве источника энергии. Обнаруженные в структуре аборигенного консорциума «силикатные» бактерии, не способные выщелачивать металлы самостоятельно, в сочетании с АХБ способствуют значительному повышению показателей биовыщелачивания. Результаты исследований свидетельствуют о формировании в отвальных продуктах топливно-энергетического комплекса устойчивых специфических микробиоценозов, которые могут быть источниками получения высокоактивных штаммов для использования в биотехнологических процесах извлечения металлов.

Ключевые слова: аборигенное сообщество, от валы, биовыщелачивание.

i Надоели баннеры? Вы всегда можете отключить рекламу.