Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula
Microorganisms, superresistant to toxic metals, have been revealed in a geographical zone of internal island shelf of Antarctic peninsula (30x60 km) on islands Darboux, Lippmann, Berthelot, Cruls, Barchans, Jalour, Uruguay, Scua, Three pigs, Winter, Grotto, Galindez, Pitermann, Booth, capes Perez, R...
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| Cite this: | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula / A.B. Tashyrev, P.V. Rokitko, N.A. Matvieieva, A.A. Tashyreva, V.A. Romanovskaya // Український антарктичний журнал. — 2009. — № 8. — С. 276-283. — Бібліогр.: 5 назв. — англ. |
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| author | Tashyrev, A.B. Rokitko, P.V. Matvieieva, N.A. Tashyreva, A.A. Romanovskaya, V.A. |
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| citation_txt | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula / A.B. Tashyrev, P.V. Rokitko, N.A. Matvieieva, A.A. Tashyreva, V.A. Romanovskaya // Український антарктичний журнал. — 2009. — № 8. — С. 276-283. — Бібліогр.: 5 назв. — англ. |
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| description | Microorganisms, superresistant to toxic metals, have been revealed in a geographical zone of internal island shelf of Antarctic peninsula (30x60 km) on islands Darboux, Lippmann, Berthelot, Cruls, Barchans, Jalour, Uruguay, Scua, Three pigs, Winter, Grotto, Galindez, Pitermann, Booth, capes Perez, Rasmussen, Tuxen (coast of Antarctic peninsula), and also on distant from them island King-George. The highest number of metalresistant microorganisms is revealed on the medium containing 1000 ppm of Cr(VI) - up to 5,0х10⁵ cells/g of soil and on the medium containing 250 ppm of Ni²⁺ – 3,0-5,0x10⁵ cells/g of soil.
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276
OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE
INTERNAL SHELF OF THE ANTARCTIC PENINSULA
Tashyrev A.B., Rokitko P.V., Matvieieva N.A., Tashyreva A.A., Romanovskaya V.A.
Zabolotny Institute of microbiology and virology of NAS of Ukraine, Ukraine, D 03680, Kiev, str.,
154. E-mail: tach@i.com.ua
Abstract. Microorganisms, superresistant to toxic metals, have been revealed in a geographical zone of internal
island shelf of Antarctic peninsula (30x60 km) on islands Darboux, Lippmann, Berthelot, Cruls, Barchans,
Jalour, Uruguay, Scua, Three pigs, Winter, Grotto, Galindez, Pitermann, Booth, capes Perez, Rasmussen, Tuxen
(coast of Antarctic peninsula), and also on distant from them island King-George. The highest number of
metalresistant microorganisms is revealed on the medium containing 1000 ppm of Cr(VI) - up to 5,0x10 cells/g
2+ 5 of soil and on the medium containing 250 ppm of Ni – 3,0-5,0x10 cells/g of soil. Critical concentrations of toxic
2+ 2+ 2+ 2+metals for the Antarctic microorganisms are determined: Hg - 100 ppm, Cu and Ni - 500 ppm, Co - 1000
ppm. For chromium this parameter is higher than 2000 ppm. For the analysis of results of frequency of
metalresistant microorganisms occurrence used the factor representativeness (occurrence) C suggested by us oc
which allows to characterize in a complex geoobjects since connects two parameters: total of microorganisms and
amount metalresistant microorganisms. Factor C for nickelresistant and copperresistant microorganisms is oc
practically identical on the investigated geoobjects. High C are found out on cape Rasmussen and on island oc
Winter, and the lowest - on island Lippmann. Thereby, we have shown for the first time, that resistance of
microorganisms to ultrahigh concentration of toxic metals is widespread phenomenon in soils on an internal
island shelf of the Western Antarctic Region. This conclusion is based on: a) superresistancies and polyresistance
2+ 2+ 2+ 2+of the Antarctic microorganisms to toxic metals (Hg , Cu , Cr(VI), Ni , Co ); b) high frequency of occurrence
and high amount of metalresistant microorganisms in the Antarctic soil; c) occurrence of resistant to toxic metals
microorganisms in all investigated soil samples in a zone of an internal island shelf of Antarctic peninsula in the
extent 30x60 kms.
Key words: Antarctic Region, microorganisms, toxic metals, resistance, polyresistance.
1. Introduction
The phenomenon of resistance of the Antarctic microorganisms to toxic metals - one of the
least studied aspects of the Antarctic microbiology. Only in a few publications the data concerning
resistance to metals of water and ice bacteria are resulted (De Souza et al. 2006). Earlier we had been
found out microorganisms resistant to toxic metals in the Antarctic samples selected on
biogeographical polygon of island Galindez, (Tashyrev A.B. et al., 2007). At the same time, studying
distribution resistant to toxic metals microorganisms in other geographical zones of Antarctic Region
is of interest. In this connection, the purpose of investigation was studying of metalresistant
microorganisms' distribution on the islands of an internal shelf and on the coast of Antarctic peninsula
(30x60 km zone).
2. Methods and materials
Research objects were aerobic heterotrophic microorganisms isolated from native and frozen
(-20°Ñ) samples of soils. Samples were selected during 13-th Antarctic expedition on geographical
objects (further in the text the term "geoobjects" is used) internal island shelf of Antarctic peninsula.
Preparation of solutions of toxic metals. Toxic metals brought in as solutions of salts
CuCI x2H O, K CrO , Hg(NO ) Ni(NO ) x6H O è CoCl x6H O. Necessary concentration of toxic 2 2 2 4 3 2, 3 2 2 2 2
2+ 2+ 2+metals in a nutrient medium obtained by dilution of initial solutions: 20 g/l for Cu , Co , Ni , Ñr(VI)
5
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BIOLOGICAL RESEARCH
277
2+and 4 g/l for Hg (recalculation on a cation of metal). Nitrate of mercury obtained by dissolving 1 g
metal mercury in 5,0 ml concentrated HNO . A solution evaporated on a water bath, and then Hg(NO ) 3 3 2
dissolved in distilled water and lead up volume up to 1 liter. Solutions of metal ions sterilized during
20 min on boiling water bath, then cooled up to 30°Ñ and brought in sterile conditions in a nutrient
medium up to necessary final concentration.
Revealing of metalresistant microorganisms in samples was carried out on a liquid nutrient
medium by standard methods. 0,1 g of sample placed in a tube which contained 10 ml NB medium
(Nutrient Broth, HiMedia Laboratories Pvt. Ltd, the USA) with toxic metals and cultivated in
stationary conditions at +20°Ñ. Used such concentration of toxic metals in recalculation on a cation
2+ 2+ 2+ 2+(ppm): Hg - 100 and 200; Cu - 500 and 1000; Ni - 1000 and 2000; Co - 1000 and 2000, Cr (VI) -
2000 and 5000. Presence of growth of microorganisms was determined on optical density of a culture
liquid through with 3-14 day. The absence of growth of sensitive to metals strain Escherichia coli
UCM B-906 served as the control of a toxicity of medium with metals.
Number of metalresistant and total amount of chemoorganotrophic microorganisms in
researched samples determined by method of inoculation of consequent dilutions on agarized
medium. To grinded in a porcelain mortar samples (1 g) added a physiological solution (10 ml) and
mixed (1 hour). Then tenfold dilutions were inoculated on agarized medium NA (Nutrient Agar, firm
2+ 2+HiMedia Laboratories Pvt. Ltd, the USA) which contained toxic metals (ppm): Hg - 50 and 100, Cu
2+ 2+- 250 and 500, Ni - 250 and 500, Co - 500 and 1000, Cr (VI) - 1000 and 2000. The same dilutions
were inoculated on an agarized media without metals for estimation of total amount of
îmicroorganisms. A cultivation carried out at +20 Ñ. Colonies counted up after 7-10 days.
Morphological types of colonies counted up in plates where their total number did not exceed 50. As
the basic criteria for definition of morphological types of colonies on the base of such criteria: shape,
diameter, consistence, pigmentation, synthesis of water soluble pigment, presence of extracellular
slime, and presence of air and substrate mycelium. Single colonies selected and transferred on the
same medium for their storage in laboratory conditions.
Occurrence frequency of metalresistant microorganisms determined by factor of
representativeness C suggested by us. This parameter calculated as a ratio of metalresistant oc
microorganisms number in 1 g of sample and total amount of microorganisms selected on medium
without metals: C = (N : N ) 100 %,oc met con
where C is a representativeness factor; N – number of metalresistant microorganisms on a medium oc met
with metal;N - total amount of microorganisms on a medium without metals. con
Polyresistance of microorganisms to metals determined by inoculation of representative
2+ 2+ strains on agarized medium, each of which contained on one toxic metal (ppm):Hg – 50; Cu – 250;
2+ 2+ Ni – 250; Co – 500; Cr(VI) – 1000.
3. Results and their discussion
Occurrence of metalresistant microorganisms on islands of an internal shelf of Antarctic
peninsula. The investigated zone in the extent of 60 kms (from the North on the South) and 30 kms
(from the West on the East) includes islands in the strait Penola and capes on the Antarctic peninsula.
18 biogeographical objects are investigated: islands Darboux, Galindez, Skua, Barchans, Uruguay,
Jalour, Petermann, Berthelot, Cruls, Booth, Grotto, Three pigs, Winter, Lippmann, capes Rasmussen,
Tuxen and Perez. Samples also were selected on island King-George which is considerably distant
from the basic investigated zone.
For revealing metalresistant microorganisms screening soil samples on a liquid nutrient
medium is carried out. It is established, that in a researched zone of Antarctic Region microorganisms
2+superresistant to Cu (500 ppm) and Cr(VI) (2000 ppm) (fig. 1) are widely ocurred. The attention that
resistant microorganisms have been found out in 50-70 % of samples even at presence in medium
5000 ppm of chromium, and/or 1000 ppm of copper, and/or 1000 ppm of cobalt pay to itself. Such data
testify to extremely high level of resistance of the Antarctic microorganisms to metal characterized by
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
278
2-high reductive-oxidative potential, that are very toxic because of oxidative properties, CrO 4
’ 2- 2+ (Å =+555 ìV for reaction of reduction CrO to Cr(ÎÍ) )and also to Cu - to metal which combines î 4 3
2+ properties as the metal-substituent so the metal-oxidizer (Å =+440 ìV for reaction of reduction Cuî
to Cu Î), that was discussed in previous work (Tashyrev A.B. et al., 2008). The most toxic for soil 2
2+ 2+microorganisms appeared such metals as Hg and Ni . As a whole, microorganisms, superresistant to
toxic metals, have been revealed on all investigated geoobjects of Antarctic Region. Therefore further
we determined the quantitative characteristics of metalresistant microorganisms in some regions of
Antarctic Region.
’
Fig. 1. Number of geoobjects (%) an internal island shelf of Antarctic peninsula in which
microorganisms resistant to the specified concentration of toxic metals was revealed.
The quantitative account of metalresistant microorganisms has been carried out in soil of four
geoobjects (islands Darboux, Lippmann, Winter and cape Rasmussen) by a method of inoculation of
microorganisms from samples on agarized medium which contains the different toxic metals. The
highest amount of metalresistant microorganisms is revealed on the medium with 1000 ppm of
5 5chromium - up to 5,0x10 cells/g of soil, and on the medium with 250 ppm of nickel – 3,0-5,0x10
cells/g of soil (tab. 1).
Table 1
Number of the metalresistant microorganisms revealed in soil samples
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
279
At higher concentration of these metals in the medium the amount of cells is less, though on
separate geoobjects (for example, island Winter) the amount of resistant to chromium of
microorganisms practically was not decreased. Thus, the Antarctic soil microorganisms showed
significant resistance to high concentration Cr(VI).
At the same time, the amount of microorganisms on the medium with 50 ppm of mercury was
2in limits 1,0-3,0x10 , and already at concentration of 100 ppm of mercury resistant microorganisms
have been found out only on island Lippmann (tab. 1). For presentation in figure 2 the number of
metalresistant microorganisms which is revealed in soil of island Darboux is resulted.
In these experiments critical concentration of toxic metals for the Antarctic microorganisms (in
agarized medium) are determined: mercury - 100 ppm, copper - 500 ppm and cobalt - 1000 ppm (tab.
1). For chromium and nickel this parameter is not determined, as at used concentration of these metals
3 5(fig. 2) the significant amount of microorganisms (from 10 up to 10 cells/g of soil) is revealed. The
established critical concentration of toxic metals for the Antarctic microorganisms are higher 1-2
order than for microflora of other regions that allows to count isolated microorganisms perspective for
use in biotechnology of purification from metals of industrial waste waters.
2+ 2+ 2+ 2+ Fig. 2. Number of resistant to Co , Ni , Cu , Cr(VI), Hg microorganisms in 1 g of soil of
island Darboux (K - number of microorganisms on the medium without metals).
Determination of occurrence frequency (factor of a representativeness) of metalresistant
microorganisms. The total number of heterotrophic microorganisms (on the medium without metals)
7 was in limits 0,8-2,0x10 cells/g of soil (tab. 1). Using this parameter and number of metalresistant
Antarctic microorganisms (tab. 1) we calculated a factor of a representativeness (occurrence) of
metalresistant microorganisms - C (tab. 2). oc
Factor C allows characterizing in a complex the phenomenon of metalresistance of oc
microorganisms on the investigated geoobjects. It is shown, that the factor of C nickelresistant oc
microorganisms is practically identical on all investigated geoobjects. Similar factors are obtained and
for copperresistant microorganisms, but with lower value (tab. 2). This law is distinctly visible in
figure 3. The highest C values are found out for samples of cape Rasmussen and on island Winter, and oc
the lowest - on island Lippmann if to compare among themselves geoobjects (fig. 3).
As a whole, factor C gives representation about relative occurrence frequency of oc
metalresistant microorganisms and allows to characterize in a complex geoobjects since is a
derivative of two characteristics: total number of microorganisms in a sample and amounts of
metalresistant microorganisms among them.
Thus, obtained results of the microbiological analysis of soil samples have shown, that in the
investigated biogeographical zone on all 18 geoobjects of an island shelf microorganisms, resistant to
2+ 2+ 2+ 2+high concentration of toxic metals (Hg , Cu , Ni , Co , Cr(VI)) are found out.
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
280
Table 2
Factor of representativeness of metalresistant microorganisms
on four investigated geoobjects of Antarctic Region
The note: "-" - did not calculate, since on these geoobjects microorganisms, resistant to the
specified metals have not been found out.
Fig. 3. Factors of representativeness (C ) of the Antarctic metalresistant microorganisms. oc
Figures above columns specify metal and its concentration in the medium (ppm): 1 – Co(500), 2 –
Co(1000), 3 – Ni(250), 4 – Ni(500), 5 – Cr(1000), 6 – Cr(2000), 7 – Cu(250), 8 – Cu(500), 9 – Hg(50),
10 – Hg(100).
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
281
On the map-scheme (fig. 4) the geographical zone of an internal island shelf of Antarctic
peninsula investigated by us (30x60 km), where the investigated geoobjects are designated, is
resulted. As follows from the given scheme, on all investigated geoobjects of Antarctic Region (15
islands and 3 capes of Antarctic peninsula) have been revealed metalresistant microorganisms.
Polyresistance of the Antarctic microorganisms. As a result of the carried out microbiological
analysis of the soil samples selected on 18 geoobjects of an internal shelf of Antarctic peninsula, we
developed a collection of metalresistant microorganisms (50 strains). At 21 strains (any way chosen
from this collection) ability to grow has been investigated at the presence of different toxic metals (tab.
3). Polyresistance of microorganisms determined by inoculation strains on agarized medium which
2+ 2+ 2+ 2+contained on one toxic metal in such concentration (ppm): Hg – 50; Cu – 250; Ni – 250, Co – 500
è Cr(VI) – 1000. It is shown, that 5 strains show resistance to all five metals, to four metals show
resistance of 10 strains and to three metals - 5 strains (îne strain has not shown polyresistance, that is
was resistant only to one metal) (fig. 5).
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
x
282
Table 3
Polyresistancy to toxic metals of isolated Antarctic soil microorganisms
The note: R - presence of growth; S - absence of growth.
All researched microorganisms isolated on medium with mercury, appeared to be resistant as
well to copper pays attention to it. On the other hand – all copperresistant microorganisms were
resistant to mercury. These data allow suggests that, probably, the isolated strains have similar
2+ 2+mechanisms of resistance to Cu and Hg .
It is necessary note that for strains of the microorganisms isolated from samples of polygon
island Galindez (seasonal expeditions of 2001-2008), polyresistence to metals also is inherent
(Tashyrev A.B. et al., 2007).
Fig. 5. Percent of strains characterized by polyresistance to metals: 47 % of the investigated
microorganisms resistant to five metals, 23 % - to four and three metals (see tab. 3), 5 % - to one.
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
283
Thus, for the first time we have shown that resistance of microorganisms to ultrahigh
concentration of toxic metals is widespread phenomenon in soil of an internal island shelf of the
Western Antarctic Region. This conclusion is based on: a) superresistance and polyresistance of the
2+ 2+ 2+ 2+Antarctic microorganisms to toxic metals (Hg , Cu , Cr(VI), Ni , Co ); b) high frequency of
occurrence and a significant amount of metalresistant microorganisms in the Antarctic soils; c)
occurrence of resistant to toxic metals microorganisms in all investigated soil samples in a zone of an
internal island shelf of Antarctic peninsula in the extent 30×60 kms.
The Reference
1. Tashirev O.B., Galinker E.V., Andreyuk E.I. Thermodynamic forecasting of redox-
2+ 2– 2+interaction of microorganisms with metals-oxidizers (Hg , CrO and Cu )// Reports of the National 4
Academy of sciences of Ukraine. - 2008. - ¹ 4. - P. 166-172 (in Russian).
2. Tashyrev O.B., Matvieieva N.A., Romanovskaya V.A., Tashyreva A.O., Rokitko P.V.
Polyresistance and superresistance of Antarctic microorganisms to heavy metals//Reports of the
National Academy of sciences of Ukraine. - 2007. ¹ 11. - P. 70-75 (in Russian).
3. De Souza M.J., Nair S., Loka Bharathi P.A. et al. Metal and antibiotic-resistance in
psychrotrophic bacteria from Antarctic Marine waters // Ecotoxicology. – 2006. – 15, ¹ 4. – P. 379-
384.
The information on authors
The address:
Ukraine, D 03680, Kiev, Zabolotnogo str., 154. Zabolotny Institute of microbiology and virology
of NAS of Ukraine
Tashyrev A.B. - head of department of extremophilic microorganisms biology of Zabolotny
Institute of microbiology and virology of NAS of Ukraine, Dr.Sci., senior scientific researcher,
microbiologist, biotechnologist. Ph. 8 (044) 526-32-97. E-mail: tach@i.com.ua
Rokitko P.V. - senior scientific researcher of department of extremophilic microorganisms
biology of Zabolotny Institute of microbiology and virology of NAS of Ukraine, Ph.D.,
microbiologist, ecologist. Ph. 8 (044) 526-32-97.
Matveeva N.A. - leading engineer of department of extremophilic microorganisms biology of
Zabolotny Institute of microbiology and virology of NAS of Ukraine, Ph.D., microbiologist,
biotechnologist.
Tashyreva A.A. - leading engineer of department of extremophilic microorganisms biology of
Zabolotny Institute of microbiology and virology of NAS of Ukraine, microbiologist.
Romanovskaya V.A. - leading scientific researcher of department of extremophilic
microorganisms biology of Zabolotny Institute of microbiology and virology of NAS of Ukraine,
Dr.Sci., professor, microbiologist, ecologist.
Tashyrev A.B.: OCCURENCE OF METALRESISTANT MICROORGANISMS ON ISLANDS OF THE INTERNAL...
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| id | nasplib_isofts_kiev_ua-123456789-128569 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1727-7485 |
| language | English |
| last_indexed | 2025-12-07T18:22:18Z |
| publishDate | 2009 |
| publisher | Національний антарктичний науковий центр МОН України |
| record_format | dspace |
| spelling | Tashyrev, A.B. Rokitko, P.V. Matvieieva, N.A. Tashyreva, A.A. Romanovskaya, V.A. 2018-01-11T19:43:27Z 2018-01-11T19:43:27Z 2009 Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula / A.B. Tashyrev, P.V. Rokitko, N.A. Matvieieva, A.A. Tashyreva, V.A. Romanovskaya // Український антарктичний журнал. — 2009. — № 8. — С. 276-283. — Бібліогр.: 5 назв. — англ. 1727-7485 https://nasplib.isofts.kiev.ua/handle/123456789/128569 579.695 Microorganisms, superresistant to toxic metals, have been revealed in a geographical zone of internal island shelf of Antarctic peninsula (30x60 km) on islands Darboux, Lippmann, Berthelot, Cruls, Barchans, Jalour, Uruguay, Scua, Three pigs, Winter, Grotto, Galindez, Pitermann, Booth, capes Perez, Rasmussen, Tuxen (coast of Antarctic peninsula), and also on distant from them island King-George. The highest number of metalresistant microorganisms is revealed on the medium containing 1000 ppm of Cr(VI) - up to 5,0х10⁵ cells/g of soil and on the medium containing 250 ppm of Ni²⁺ – 3,0-5,0x10⁵ cells/g of soil. en Національний антарктичний науковий центр МОН України Український антарктичний журнал Біологічні дослідження Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula Article published earlier |
| spellingShingle | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula Tashyrev, A.B. Rokitko, P.V. Matvieieva, N.A. Tashyreva, A.A. Romanovskaya, V.A. Біологічні дослідження |
| title | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula |
| title_full | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula |
| title_fullStr | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula |
| title_full_unstemmed | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula |
| title_short | Occurence of Metalresistant Microorganisms on Islands of the Internal Shelf of the Antarctic Peninsula |
| title_sort | occurence of metalresistant microorganisms on islands of the internal shelf of the antarctic peninsula |
| topic | Біологічні дослідження |
| topic_facet | Біологічні дослідження |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/128569 |
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