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Nova Biotechnologica 10-1 (2010) 71

BIO-TREATMENT OF SOIL FROM
INTERNATIONAL AIRPORT IN OSTRAVA

PETER FEČKO, RADMILA KUČEROVÁ, EVA PERTILE,

LUCIE NEZVALOVÁ, NIKOLAS MUCHA,
IVA JANÁKOVÁ, VLADIMÍR ČABLÍK

VSB – Technical University of Ostrava, Faculty of Mining and Geology,

17. listopadu 15, 708 33 Ostrava - Poruba, Czech Republic (peter.fecko@vsb.cz)

Abstract: The paper deals with an examination of possible application of biodegradation in the
decontamination of soil samples from international airport in Ostrava. The laboratory biodegradation tests
were carried out with a pure bacterial culture of Pseudomonas putida, a pure laboratory culture of
Rhodococcus sp, their mixture and a mixture prepared combining their media free of bacteria. The results of
the paper imply that for biodegradation of airport pollutants is most suitable to apply a mixed bacterial
culture of Pseudomonas putida and Rhodococcus sp. The results show that the biodegradation method is
applicable for the pollution.

Keywords: biodegradation, Pseudomonas putida, Rhodococcus sp., mixed culture

1. Introduction

Biodegradation (biological decontamination) is grounded in the capacity of certain

bacterial strains to use hydrocarbons as a source of carbon and energy for their growth and
in this way, decomposition of contaminants occur all the way to harmless products - carbon
dioxide and water. In short, biodegradation is a special case of degradation during which
decomposition of polymers takes place due to the action of biological factors. It makes part
of natural processes taking place in water and soil. For example, there is spontaneous
degradation of biologically degradable oil substances at contamination of soil by oil
substances. However, the process is slow and meanwhile contamination may spread into
the surroundings. In the locality some resistant substances remain. In order to speed up the
rate of degradation, it was necessary to make the process more intense and to remove
resistant substances bacterial mixtures may be utilized.

The ability of microorganisms to degrade hydrocarbons has been known since 1895,
when Miyoshi described growth of yeast on paraffin and shortly after the capacity of
bacteria to make use of methane as a source of carbon was discovered. Gradually, it was
demonstrated that they are able to decompose practically all components of crude oil and
many other hydrocarbons. At present, over 200 types of microorganisms have been
described that are able to degrade hydrocarbons. Some are able to make use of one
hydrocarbon only (e.g. methane), but no microbial strain is known to degrade a whole
range of hydrocarbons present in crude oil, for example. Therefore, these are rather
microbial associations that participate in degradation (NOVOTNÝ, 2005).

The objective of the paper was to examine the application of bacterial leaching to the
decontamination of soil samples from the Leos Janacek Airport in Ostrava – these samples
were taken from two sampling points.

72 Fečko, P. et al.

2. Materials and methods

2.1 Leos Janacek Airport in Ostrava

The first mention of air traffic at the territory of the present airport is from 1939 when

German Luftwaffe built a field aerodrome to attack Poland. The modern history began in
1956 when the current airport began to be constructed. Before 1989, the airport was used
mainly for the needs of the air force. Civil aviation was ensured by CSA, namely for
domestic flights, rarely for international ones. A significant turning point was the year of
1993 when the military traffic was terminated at the airport. On 13 December 2006 the
airport was ceremonially christened after the composer of Leos Janacek and a new
departure hall was put into operation. With regard to its excellent technical parameters,
a pronounced development of this traffic junction is expected in the future (see
references).

Several oil leaks have occurred at the airport and Figure 1 and Figure 2 show an oil
interceptor Lapol D and Lapol 2, which intercepts leaks of hazardous substances. Table 1
summarizes leaks of hazardous substances since 2005. At the same time, it gives
substances that penetrated into the environment and the materials by used for their
elimination.

Table 1. Leaks of oil substances at the airport.

Date Leak locality Substance
Qty of
leaked

subst. [l ]

Material
used

for disposal

Leak into
sewer
system

23.6.2005 Central passenger terminal JET A – 1 200
Vapex,
Cansorb NO

21.6.2006 Central passenger terminal JET A – 1 Not determ. Cansorb NO

28.7.2006 Central passenger terminal Hydraulic oil Not determ. Cansorb NO

12.9.2006 Central passenger terminal JET A – 1 Not determ. Cansorb NO

18.9.2006 Taxiway JET A – 1 50

Vapex,
Cansorb, wa
ter, surface-
active agent

27.9.2006 Northern stand JET A – 1 30 Cansorb NO

29.3.2007 Central passenger terminal JET A – 1 30 Cansorb NO

28.3.2008
Bunkers of

airport
propellants

Oil products –
closely

unspecified
Not determ.

Vapex,
sorption

layer,
absorption

heaps,
sewer seal

YES

Nova Biotechnologica 10-1 (2010) 73

It is apparent from the table that accidents with environmental impact prevail,
accompanied by leaks of aviation turbine fuel JET A-1.

Fig. 1. Lapol D – leaks of oil products. Fig. 2. Lapol 2.

2.2 Characteristics of drawn samples

The soil samples were taken directly from the airfield of the Leos Janacek Airport
in Ostrava – Mosnov - see Figure 3.

Fig. 3. View of the sampling point.

The mineralogical analyses were implemented in the laboratories of the Institute of

Geological Engineering at Mining College - Technical University of Ostrava by means of
an X-ray diffraction. The results of the mineralogical analyses of sample 1 (Figure 4) imply
that the samples contain about 18 % of amorphous phase, majority of quartz - about 57 %
and followed by calcite, chlorite, muscovite, orthoclase and albite.

Fig. 4. Mineralogical analysis of the sample No.1.

74 Fečko, P. et al.

The results of the mineralogical analyses of sample 2 (Figure 5) imply that the samples
contain about 30 % of amorphous phase, majority of quartz - about 42 %. .

Fig. 5. Mineralogical analysis of the sample No.2.

2.3 Characteristics of bacterial cultures and the method of laboratory tests

For biodegradation of the samples, pure bacterial cultures of Pseudomonas putida and
Rhodococcus sp were used. The bacterial cultures are shown in Figures 6 A, B (FEČKO et
al., 2004).

Fig. 6. Pseudomonas putida (A) and Rhodococcus sp. RHA1 (B) (FEČKO et al., 2004).

The culture media were the liquid medium of M1 for Pseudomonas putida and medium
of M96 for Rhodococcus sp.

The laboratory experiments were carried out with pure bacterial cultures of
Psedomonas putida and Rhodococcus sp., mixed culture and bacterial medium made of 50
% MI medium and 50 % of M96 medium. The experiments were carried out in the
laboratories of the Institute of Environmental Engineering at VSB-TU Ostrava, where 28-
day bacterial degradation took place. Each sample was placed into a 2 l glass beaker.
Aeration was secured by means of aquarium pumps placed into the beakers. The necks of
the beakers were sealed with a foil and then the beakers were moved into the chemical
hood. In the course of 4-week degradation the volume in the beakers was regularly filled
with distilled water as gradual evaporation occurred. Having finished the experiment, the
samples were filtered, dried and sent to further chemical analyses into the Brown Coal
Research Institute in Most.

Nova Biotechnologica 10-1 (2010) 75

3. Results and discussion

The results of laboratory biodegradation tests after one-month biodegradation with
applied pure bacterial cultures and mixed culture are stated in Tables 2, 3. The tables imply
that in the course of biodegradation tests, gradual degradation of harmful substances
content from the sample occurred. For biodegradation the following pure bacterial cultures
were used: Pseudomonas putida - PP, Rhodococcus sp. - R, mixed culture Rhodococcus
sp. and Pseudomonas putida - R+PP, and a check sample from media mixtures - K.

Table 2. Course of degradation the selected contaminants by means of Rhodococcus - R, Pseudomonas putida -
PP and mixed culture PP+R, check test – K.

Evaluation of the biodegradation test of sample 1

Input R Removal degree PP
Removal
degree PP+R

Removal
degree

Check
test - K

Removal
degree Parameter

mg/kg mg/kg % mg/kg % mg/kg % mg/kg %

NEL*) 196 127 35.2 120 38.78 72 63.27 89 54.59

anthracene 11.4 1.3 88.6 1.03 90.96 2.22 80.53 4.28 62.46

benzo(a)
anthracene 65.8 8.2 87.54 5.83 91.14 12.88 80.43 29.09 55.79

benzo(b)
fluoranthene 67.2 11.51 82.87 7.74 88.48 14.61 78.26 40.22 40.15

benzo(k)
fluoranthene 61.2 54.16 11.5 5.97 90.25 11.81 80.7 27.36 55.29

benzo(a)
pyrene 105 102.29 2.58 3.71 96.47 5.78 94.5 17.66 83.18

benzo(ghi)
perylene 56.5 49.76 11.93 3.09 94.53 6.13 89.15 14.31 74.67

Fenantrene 208.8 32.03 84.66 22.69 89.13 46.36 77.8 114.38 45.22

fluoranthene 264 24.05 90.89 18.88 92.85 36 86.36 96.96 63.27

chrysene 86.9 0.32 99.63 0.72 99.17 16.41 81.12 42.57 51.01

Indeno
(1,2,3-d)pyrene 18.7 0.1 99.47 11.98 35.94 4.44 76.26 18.67 0.16

naftalene 12.3 1.35 89.02 0.95 92.28 2.14 82.6 3.71 69.84

pyrene 230.9 4.08 98.23 12.45 94.61 25.16 89.1 64.17 72.21

Σ PAH 1188.7 289.15 75.68 95.04 92 183.94 84.53 473.38 60.18

PCB 28 0.01 0.01 0 0.01 0 0.01 0 0.01 0

PCB 52 0.01 0.01 0 0.01 0 0.01 0 0.01 0

PCB 101 0.01 0.01 0 0.01 0 0.01 0 0.01 0

PCB 118 0.02 0.01 50 0.01 50 0.01 50 0.02 0

PCB 138 0.02 0.02 25 0.02 25 0.01 45 0.02 0

PCB 153 0.06 0.02 57.89 0.04 36.84 0.02 59.65 0.04 24.56

PCB 180 0.02 0.01 50 0.02 15 0.01 50 0.01 50

Σ PCB 0.15 0.09 39.46 0.11 26.53 0,08 42.86 0.12 16.33

*) NEL – hydrocarbons C10 – C40

76 Fečko, P. et al.

It is apparent from the results of four-week biodegradation test that the most
suitable application for the sample 1 is that of the pure bacterial cultures of
Pseudomonas putida, where the degradation of contaminants of PAH was 92 %.

In terms of degradation of PCB the best was the application of mixed culture, i.e.
42.9 %. In this case, the efficiency of the mixed bacterial culture was very positive as
is visible from the following removed quantities: 63.3 % of NEL, 84.50 % of PAH and
42.9 % of PCB.

Table 3. Course of degradation the selected contaminants by means of Rhodococcus - R, Pseudomonas putida -
PP and mixed culture PP+R, check test – K.

Evaluation of the biodegradation test of sample 2

Input R Removal degree PP
Removal
degree PP+R

Removal
degree

Check
test -

Removal
degree Parameter

mg/kg mg/kg % mg/kg % mg/kg % mg/kg %

NEL 150 100 33.3 60 60 35 76.6 120 20

antracene 0.7 0.49 30 0.07 90 0.1 85.71 0.12 82.86

benzo(a)
antracene 2.36 1.89 19.92 0.33 86.02 0.44 81.36 0.48 79.66

benzo(b)
fluoranthene 5.2 4.95 4.81 0.6 88.46 0.69 86.73 0.71 86.35

benzo(k)
fluoranthene 3.1 2.99 3.55 0.43 86.13 0.54 82.58 0.59 80.97

benzo(a)
pyrene 5.31 4,67 12.05 0.53 90.02 0.59 88.89 0.74 86.06

benzo(ghi)
perylene 3.5 1.16 66.86 0.21 94 0.27 92.29 0.28 92

fenantrene 2.65 2.45 7.55 0.86 67.55 0.14 94.72 1.45 45.28

fluoranthene 5.7 0.27 95.26 0.19 96.67 0.41 92.81 0.58 89.82

chrysene 2.7 0.13 95,19 0.25 90.74 0.02 99.26 0.02 99.26

indeno(1,2,3-cd)
pyrene 6.99 6.66 4.72 2.39 65.81 0.11 98.43 0.56 91.99

naftalene 0.09 0.07 22.22 0.07 22.22 0.07 22.22 0.07 22.22

pyrene 4.75 2.69 43.37 0.6 87.37 0.17 96.42 1.3 72.63

Σ PAH 43.05 28.42 33.98 6.53 84.83 3.55 91.75 6.9 83.97

PCB 28 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

PCB 52 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

PCB 101 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

PCB 118 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

PCB 138 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

PCB 153 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

PCB 180 <0.01 <0.01 - <0.01 - <0.01 - <0.01 -

Σ 7 PCB <0.07 <0.07 - <0.07 - <0.07 - <0.07 -

*) NEL – hydrocarbons C10 – C40

Nova Biotechnologica 10-1 (2010) 77

It is apparent from the results of four-week biodegradation test that the most
suitable application for the sample 2 is that of mixed bacterial cultures of
Pseudomonas putida and Rhodococcus, where the degradation of contaminants of PAH
was 91.75 %. The amount of PCB was lower as detection limit.

In this case, the efficiency of the mixed bacterial culture was very positive as is
visible from the following removed quantities: 76.6 % of NEL, 84.83 % of PAH and 0
% of PCB.

4. Conclusions

The objective of the paper was to examine the application of biodegradation in the

decontamination of soil sample from the Leos Janacek Airport in Ostrava.
For the laboratory biodegradation tests soil samples from the airport of the Leos

Janacek in Ostrava – Mosnov was used. The laboratory biodegradation tests were
implemented with pure bacterial culture of Pseudomonas putida, pure bacterial culture
of Rhodococcus sp, their mixture and mixture made combining their media free of
bacteria.

The efficiency of the biodegradation of sample 1 after one-month leaching with
pure bacterial culture of Pseudomonas putida (PP) was 38.8 % for NEL, 92% for
PAH, 26.5 % for PCB, by means of pure bacterial culture of Rhodococcus sp. (R) was
35.2 % for NEL, 75.7 % for PAH, 39.5 % for PCB, by means of mixed bacterial
culture it was 63.3 % for NEL, 84.5 % for PAH, and 42.9 % for PCB.

The efficiency of the biodegradation of sample 2 after one-month leaching with
pure bacterial culture of Pseudomonas putida (PP) was 60 % for NEL, 84.5% for
PAH, 0 % for PCB, by means of pure bacterial culture of Rhodococcus sp. (R) was
33.3 % for NEL, 28.5 % for PAH, <0.07 % for PCB, by means of mixed bacterial
culture it was 76.6 % for NEL, 91.8 % for PAH and 0 % for PCB.

The paper results imply that the laboratory sample 1 biodegradation efficiency with
the selected contaminants ranged from 35.2 – 63.3 % for NEL, 60.2 - 92 % for PAH
and 16.3 – 42.9 % for PCB.

The paper results imply that the biodegradation efficiency of laboratory sample 2
with the selected contaminants ranged from 33.3 – 76.6 % for NEL, 33.98 – 91.75 %
for PAH. The amount of PCB was lower as detection limit.

The best efficiency was obtained in the laboratory biodegradation of PAH.
Intermediate efficiency was reached with biodegradation of NEL and PCB. The
highest amount of NEL and PCB were removed by mixed bacterial culture (PP+R).
For soil biodegradation, it is thus the most suitable to apply the mixed bacterial culture
of Pseudomonas putida and Rhodococcus sp.

The results demonstrate that for the given type of contamination the method of
biodegradation is suitable.

Acknowledgement: The work was carried out within the VaV SP/2f2/98/07 Project solution
“Research in the Utilization of Waste as Substitutes for Primary Raw Material Sources” and
supported by the Ministry of Environment (MoE) of the CR.

78 Fečko, P. et al.

References

NOVOTNÝ, C.: Biodegradace a biotechnologie, Ostravska univerzita, Ostrava, 2005.
96 pp.

Basic information on the airport [online]. 2009 [cit. 2009-03-02]. Available from
WWW: <http://www.airport-ostrava.cz/cz/page-zakladni informace>.

History of the airport [online]. 2008 [cit. 2009-03-02]. Available from WWW:
<www.airport-ostrava.cz/cz/page-historie-vznik-vyvoj>.

Technical data on the airport [online]. 2008 [cit. 2009-03-02]. Available from WWW:
<http://www.airport-ostrava.cz/cz/page-technicke-udaje>.

Mosnov municipality [online]. 2009 [cit. 2009-03-02]. Available from WWW:
<http://www.mosnov.cz/index.php?lang=1&str=002>.

FECKO, P. et al.: Environmentalní biotechnologie, Ostrava, VSB- TU Ostrava, 2004.
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