Atmospheric Pollution Research xxx (2016) 1e8

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Original article

Effects of the urban environmental conditions on the physiology

of lichen and moss
Gintare_ Sujetoviene_ *, Vilma Galinyte_
Department of Environmental Sciences, Vytautas Magnus University, Vileikos 8, Kaunas LT-44404, Lithuania

a r t i c l e i n f o a b s t r a c t

Article history: The lichen Evernia prunastri (L.) Ach. and the moss Ptilium crista-castrensis (Hedw.) DeNot were exposed
Received 12 October 2015 in three different urban sites along with the different pollution loads in order to evaluate the environ-
Received in revised form mental quality. Physiological changes after the exposition were assessed by evaluating the content of
19 February 2016
photosynthetic pigments, chlorophyll uorescence, membrane lipid peroxidation and integrity of cell
Accepted 23 February 2016
Available online xxx
membranes. The differences in response of uorescence, chlorophylls contents and injuries of cell
membranes were observed between the two species and between the sites. E. prunastri showed a great
capacity to sustain the photosynthesis processes in the urban environment, while this capacity was very
Keywords:
Air quality
low in the transplanted moss P. crista-castrensis. The levels of malondialdehyde (MDA) indicated a
Biomonitoring signicantly higher oxidative stress in the transplants at urban and residential sites. The integrity of cell
Lichen membranes in moss samples was also more damaged than in lichen.
Moss Copyright 2016 Turkish National Committee for Air Pollution Research and Control. Production and
Urban hosting by Elsevier B.V. All rights reserved.

1. Introduction of contaminants (Minger and Krahenbuhl, 1997; Conti and

The increased number in human population is leading to
changes in the level of urbanization, which results in increasing
levels of pollutants in the environment (Fenger, 1999; Duh et al.,
2008). In such an environment, the main pollutants are nitrogen
and sulphur oxides, heavy metals, mainly resulting from transport
emissions and industrial activities. The increased concentrations of
pollutants have a negative impact on health of living organisms
including human (Brunekreef and Holgate, 2002; Gurjar et al.,
2010). It is difcult to monitor the quality of the air due to the ex-
istence of different substances as well as temporal and spatial
variation in their concentrations. In order to monitor atmospheric
pollution, the biomonitoring methods based on using living or-
ganisms has attained an increasing attention in recent decades
(Sujetoviene, 2015).
Cryptograms are directly dependent on the atmospheric depo-
sition as the source of nutrients, and are sensitive to changes in
their concentrations due to their physiology. Researchers have
emphasized the signicance of lichens and mosses as biomonitors
* Corresponding author. Tel.: 370 37 352937; fax: 370 37 327916.
_
E-mail address: g.sujetoviene@gmf.vdu.lt (G. Sujetoviene).
Peer review under responsibility of Turkish National Committee for Air
Pollution Research and Control.
Cecchetti, 2001; Szczepaniak and Biziuk, 2003). It has been
shown that mosses and lichens, in spite of all disadvantages, are
good tools for the air pollution monitoring, however, the best re-
sults could be achieved if both of them are used together, because of
the differences in their metal uptake and retention (Szczepaniak
and Biziuk, 2003).
Biomonitors can serve as the means to get a spatial and tem-
poral distribution of pollutants deposition. The comparative use of
lichens and mosses in order to observe the source of pollution has
been demonstrated by several authors. A larger part of studies has
intended to compare the capacity to accumulate airborne heavy
metals (Chiarenzelli et al., 2001; Bargagli et al., 2002; Szczepaniak
and Biziuk, 2003; Culicov and Yurukova, 2006; Icel and Cobanoglu,
2009; Aprile et al., 2010; Spagnuolo et al., 2011; Balabanova et al.,
2014; Gerdol et al., 2014; Kar et al., 2014) as well as organic com-
pounds (Augusto et al., 2013; Bajpai et al., 2013b). Trace elements
showed a higher accumulation in the moss than in the lichen
(Coskun et al., 2009; State et al., 2012). In most cases, the moss was
considered to be a better choice than the lichen for the bio-
monitoring of atmospheric deposition of metals, and it is argued
that mosses may generally be more suited for this purpose as
compared to lichens. The comparison with the measured and
modelled N deposition data shows that lichens are particularly able
to reect the local N deposition (Boltersdorf et al., 2014). It was

http://dx.doi.org/10.1016/j.apr.2016.02.009
1309-1042/Copyright 2016 Turkish National Committee for Air Pollution Research and Control. Production and hosting by Elsevier B.V. All rights reserved.

_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009
2 _ V. Galinyte_ / Atmospheric Pollution Research xxx (2016) 1e8
G. Sujetoviene,
observed that the accumulation of gaseous pollutants is strongly
inuenced by biomonitor vitality and that lichen bags are a more
reliable and effective tool for monitoring S, N and C atmospheric
depositions in urban areas as compared to moss bags due to lichens
resistance to dry and stressing conditions of an urban environment
(Vingiani et al., 2004; Spagnuolo et al., 2011). Study on comparison
of lichen abundance and data of N and trace metal accumulation in
moss tissue showed that N concentration in moss tissues decreased
along with increase in diversity of lichens (Gerdol et al., 2014).
The comparative studies on the physiological parameters of
biomonitors are scarce. Severe ultrastructural injuries were
observed in the moss Hypnum cupressiforme Hedw. and in the
lichen Pseudevernia furfuracea (L.) Zopf after the exposure period in
the urban area of Naples city (Spagnuolo et al., 2011). In this study
the moss experienced irreversible ultrastructural changes, while
the lichen, even, preserves its life and metabolism despite of the
observed several stress marks. Thus, moss is expected to exhibit
only passive mechanisms of element uptake, while lichen could
also be involved in active mechanisms of accumulation. Data
comparing the sample vitality under the different pollution loads in
urban areas showed also considerable heavier damage in the moss
H. cupressiforme than in the lichen P. furfuracea (Tretiach et al.,
2007a).
The present study compares the ability of two biomonitors e
lichens and mosses e to reect deposition in an urban environment
based on the changes in their physiological parameters. We
monitored changes in air quality based on the changes in physio-
logical parameters of bioindicators along the temporal and spatial
scale. The aim of the study was to assess the effectiveness of lichens
and mosses as biomonitors in order to reect spatial and temporal
trends in air pollution.
2. Methods
2.1. Sampling area and sampling sites
The study was performed in Kaunas city (54 530 5000 N,
23 530100 E),
an urbanized area in central Lithuania at an altitude of
48 m above sea level (Fig. 1). The area is concentrated at the
conuence of two rivers (Nemunas and Neris) and occupies about
157 km2. The population is over 311,000. The climate is warm and
humid continental with annual rainfall of 627 mm and
Fig. 1. Location of the study sites in Lithuania.
_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009
predominant winds are SE. The mean annual temperature is 6.6  C.
The highest pollution level is characteristic of the city centre area,
where the main sources of air pollution are mobile sources. The
industrial activities are mainly located in the north-east part of the
city.
Three sites were chosen for the lichen transplantation studies
(Fig. 1):
Site 1: Suburban site (54 530 900 N, 23 500 4900 E). The study site
was a peripheral area located about 3 km east from the city
centre. This study site is characterized as having a relatively high
air quality with no factories and intensive motorways.
Site 2: Urban site (54 550 5800 N, 23 580 1400 E). This study site was
located in the northeastern part of the city, about 7 km away
from the city centre. This is an industrial area with intensive
trafc (>25,000 vehicle per day).
Site 3: Residential site (54 550 5800 N, 23 530 3100 E). The study site
was located in a densely populated area including public
buildings and shops, where local transport is the main source of
air pollution.
The sites were near the automatic monitoring stations where
pollution (NO, NO2 and SO2) data were recorded for the whole
exposure period. It was assumed that the there was no difference
between conditions of the sites because the sampled sites and
control sites belong to the same climatic region (control site e
weather: 3  C, wind SW at 5 m s1, 94% humidity; sampled sites e
weather 3  C, wind SW at 4 m s1, 93% humidity), the relief is
similar with comparable altitude (altitude of the control site e
79 m, sample sites e 77 m). Air quality of the sampled sites was
used for the comparison of the differences between these sites.
2.2. Sample collection and exposure
Samples of the moss Ptilium crista-castrensis L. and thallus of the
lichen Evernia prunastris (L.) Ach. were collected in the district of
Birstonas town forest (54 360 1000 N 241014.000 E) situated 30 km
south of Kaunas, central Lithuania, far from large urban and in-
dustrial facilities. Birstonas is a resort town and a centre of the
Nemunas Loops Regional Park. The surroundings of the town are
considered to be unpolluted with high naturality and represent the
initial (baseline) conditions of these species.
 _ V. Galinyte_ / Atmospheric Pollution Research xxx (2016) 1e8
G. Sujetoviene, 3

Collection of the samples was performed in one day e moss 2.7. Statistical analysis
carpets were collected from the ground, lichens thalli e from the
trunks of deciduous trees at 80e120 cm height from the ground. In The physiological parameters measured in the lichens and
the laboratory, remains of extraneous materials were removed mosses were subjected to one-way analysis of variance (ANOVA)
from the samples and washed three times with distilled water. between the sampling sites (P < 0.05). In the case of a signicant
The bags were made using nylon mesh (size 10  10 cm, with difference, a post-hoc comparison was run least signicant differ-
1  1 mm) and closed by a nylon string. Approximately 6 g of moss ence (LSD) method (P < 0.05). Spearman correlation coefcients
shoots and 6 g of lichen thalli were put in each bag and simulta- were also calculated between the physiological parameters
neously transplanted in the study sites. Four bags were placed at measured in the samples and the concentrations of air pollutants.
3 m above ground level in each area for 1 and 2 months (i.e., from 17 All calculations were performed with Statistica 6.0 (StatSoft. Inc.,
September to 17 October, 2013 and 18 October to 17 November Tulsa, USA) and Microsoft Ofce Excel 2010.
2013, respectively). After the exposure period (1 and 2 months), the
transplanted material from each site was divided into two sub-
3. Results
samples (several lichen thalli and stems of moss) per site and was
used for analyses. Untreated controls consisted of samples collected
3.1. Air pollution and weather conditions
and transplanted within Birstonas forest.
Before analysing the effect of exposure on transplants under the
2.3. Photosynthetic pigments
urban conditions, we analysed the temporal and spatial trends of
different air pollutants. There was no signicant difference of
Approximately 20 mg of lichen was extracted in dimethyl
concentration of pollutants between the two study periods with
sulphoxide (DMSO) (65  C for 45 min) according to Boonpragob
one exception e only NO concentration signicantly decreased
(2002). The moss samples (20 mg) were treated with DMSO at
during the second transplant exposure (P < 0.05). During exposure,
60  C for 2 h (Shakya et al., 2008). The photosynthetic pigments in
SO2 concentrations were quite low and have never exceeded the
all of the samples were extracted in triplicate. Optical densities of
limit value according to EU directive 2008/50/CE (Fig. 2). The mean
the solutions were measured at wavelengths 648.2 and 664.9 nm
values of SO2 were similar 0.92e1.10 mg m3 between sites with the
for the maximum absorption of chlorophyll a and b of moss using a
higher values during the rst exposure period. However, statisti-
spectrophotometer, respectively. The absorbance of the superna-
cally signicant differences between study sites and exposure pe-
tant was measured at 665 and 648 nm. Chlorophyll concentrations
riods were not determined (P > 0.05).
were calculated using the extinction coefcients and equation
Nitrogen oxides were the main compounds in all sites (Fig. 2).
given by Barnes et al. (1992).
NO2 was high in the urban study site (mean concentration
18.7 mg m3) with lower values in the residential and suburban sites
2.4. Integrity of cell membrane
(10.9 and 8.5 mg m3, respectively). The same tendency was char-
acteristic for NO concentration e the highest was observed in urban
The samples were several times rinsed in distilled water for
site in comparison with residential and suburban sites (2.4 and
3e5 s until their stable conductivity values were obtained. Elec-
5.3 mg m3, respectively). Therefore, the levels of nitrogen oxides
trical conductivity was measured. Then the samples were soaked in
were signicantly different between the study sites (P < 0.05).
glass bottles with 50 mL distilled water and shaken for 1 h. Elec-
trical conductivity was measured again. The samples were then
putted in the drying oven at 105  C for 24 h to obtain dry weight. 3.2. Changes in physiological parameters of transplanted samples

2.5. Membrane lipid peroxidation After one exposure month, there was no signicant difference in
chlorophyll a content in the lichen thalli comparing with the con-
Membrane lipid peroxidation in the lichen thalli and moss was trol site and the lichens transplanted in the urban environment
estimated using the content of malondialdehyde (MDA) as an in- (P > 0.05); however, after 2 months, chlorophyll a content in the
dicator of induced oxidative stress. The fragments of the samples lichens transplanted in the urban environment was signicantly
were rinsed in distilled water. Then the thalli were homogenized in lower (46.3%) in comparison with the control site (P < 0.0001,
a mortar using 0.1% (w/v) trichloroacetic acid (TCA) with the Fig. 3a). Contrary to the observed lichen response, chlorophyll a
addition of sand. The homogenate (1.5 mL) was put in Eppendorf
tubes and centrifuged at 12 000 g for 20 min. Supernatant (0.5 mL)
was collected and added to 1.5 mL of 0.5% thiobarbituric acid in 20%
TCA and then put in the glass tubes. The tubes were put in the oven
at 95  C for 30 min, cooled in an ice bath and then the solutions
were centrifuged again at 12 000 g for 10 min. The absorbance of
the supernatant was measured at 532 nm and corrected for non-
specic absorption at 600 nm.

2.6. Fluorescence

Chlorophyll a uorescence of samples was measured with a

plant efciency analyser (Handy PEA, Hansatech). Before mea-
surement of chlorophyll uorescence, thalli were dark-adapted for
15 min. Fluorescence was measured on well-wet samples, applying
a saturating ash of light of 2400 mmol s1 m2 for 1 s. The Fv/Fm
parameter (maximum quantum yield efciency of PSII) was used as Fig. 2. Air concentration of pollutants during the 1 and 2 sampling months (i.e., I
a stress indicator. period e 17.09.2013 to 17.10.2013 and II e 18.10.2013 to 17.11.2013).

_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009
4 _ V. Galinyte_ / Atmospheric Pollution Research xxx (2016) 1e8
G. Sujetoviene,

a E.prunastri 1 month b P. crista-castrensis

1,2 1,2 1 month
2 month
2 month
1,0 1,0

Chl a , mg g-1
0,8 0,8
Chl a, mg g-1

0,6 0,6

0,4 0,4

0,2 0,2

0,0 0,0
Control Suburban Urban Residential Control Suburban Urban Residential

Fig. 3. Chlorophyll a of lichen Evernia prunastri (a) and moss Ptilium crista-castrensis (b) transplanted in the urban area after 1 and 2 months.

content in the moss samples exhibited degradation after the rst
transplantation month and it was 60.3% lower than at the control
site after 2 months (P < 0.0001, Fig. 3b). The lowest concentration of
chlorophyll a was found in the moss samples collected from the
residential site.
Chlorophyll b content in E. prunastri was found to be signi-
cantly lower at the urban and residential sites (Fig. 4a). There were
no statistically different concentrations in the control and in sub-
urban site but after a longer transplantation period (2 months), the
level decreased and became similar to the urban and residential
sites. The similar trend for chlorophyll b content decrease was
characteristic of the transplanted moss P. crista-castrensis at the
suburban site (Fig. 4b). It should be noted that the content of
chlorophyll b at suburban and urban sites after one month of
transplantation was not signicantly different in comparison with
the control site (P > 0.05). The lowest chlorophyll b values in the
moss samples were detected at the residential and suburban sites
after the 2 months period. The chlorophyll b content was affected
more in the lichens as compared with the mosses in the urban sites
after all transplantation periods (51.1% and 44.5%, respectively).
Regarding the chlorophyll a uorescence, a signicantly lower
uorescence was observed in the lichen thalli exposed in the urban
environment e urban and residential sites e after 2-months
transplantation period (P < 0.01, Fig. 5). Despite a high variation
in moss uorescence after the transplantation, its signicantly
lower values were also characteristic of the samples exposed in the
urban and residential sites in comparison with the control site
(P < 0.05).
The lichens at the residential site had the highest MDA content
along with the moss samples (Fig. 6). The signicantly higher MDA
values were observed at the urban and residential sites in the lichen
thallus at the beginning of the transplantation period (after 1
month, P < 0.05), however, such a trend was not observed after 2
months (P > 0.05).
MDA content in the lichen thallus was signicantly higher in the
urban environment in comparison with the control site after the
transplantation period (P < 0.05, Fig. 6). As compared to the control
samples (P > 0.05), the oxidative stress parameter was signicantly
higher in the moss samples transplanted in urban environment
after 1 month (P < 0.05) but not after 2-months.
The values of electrical conductivity indicated an increasing
trend from suburban, urban sites to the residential area (Fig. 7).
After 2 months transplantation, all the samples in the urban envi-
ronment exhibited signicantly higher values of conductivity
reecting higher injuries of cell membranes in comparison with the
control site (P > 0.05).
3.3. The relationship between air pollution and the response of
bioindicators
Analysing the relationship between the air concentration of
pollutants and the determined changes in the transplants during
the exposure period it was calculated that each of the air pollutants
was related with some of the parameters measured in the lichen
and/or in the moss (Table 1). The signicant negative correlation of
the NO2 and content of chlorophyll a and b was characteristic for
lichen while NO and SO2 negatively affected concentration of
chlorophyll b. These atmospheric levels of nitrogen and sulphur
compounds also induced higher electrolyte conductivity. The as-
sociation of NO and content of chlorophyll a, Fv/Fm, MDA and
electrolyte conductivity was determined in moss. SO2 concentra-
tions correlated negatively with Fv/Fm measured in moss samples.

a E. prunastri 1 month b P. crista-castrensis 1 month

0.35 2 month 2 month
0.40
0.30 0.35
0.25 0.30
Chl b, mg g-1

Chl b, mg g-1

0.20 0.25
0.20
0.15
0.15
0.10
0.10
0.05 0.05
0.00 0.00
Control Suburban Urban Residential Control Suburban Urban Residential

Fig. 4. Chlorophyll b of lichen Evernia prunastri (a) and moss Ptilium crista-castrensis (b) transplanted in the urban area after 1 and 2 months.

_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009
 _ V. Galinyte_ / Atmospheric Pollution Research xxx (2016) 1e8
G. Sujetoviene, 5

Fig. 5. Chlorophyll a uorescence of lichen Evernia prunastri (a) and moss Ptilium crista-castrensis (b) transplanted in the urban area after 1 and 2 months.

a E.prunastri 1 month b P. crista-castrensis 1 month

2 month 4,0 2 month
3,5
3,5
3,0
3,0
2,5
2,5
2,0
2,0
1,5 1,5
1,0 1,0
0,5 0,5
0,0 0,0
Control Suburban Urban Residential Control Suburban Urban Residential

Fig. 6. Malondialdehyde (MDA) of lichen Evernia prunastri (a) and moss Ptilium crista-castrensis (b) transplanted in the urban area after 1 and 2 months.

Fig. 7. Electrolyte conductivity of lichen Evernia prunastri (a) and moss Ptilium crista-castrensis (b) transplanted in the urban area after 1 and 2 months.

Table 1
Spearman correlation coefcients between concentrations of pollutants and physiological parameters of lichen and moss (ns e not signicant, P > 0.05).

Lichen Moss

Chl a Chl b Fv/Fm MDA EC Chl a Chl b Fv/Fm MDA EC

NO ns 0.377 ns ns 0.417 0.447 ns 0.648 0.442 0.563

NO2 0.378 0.600 ns ns 0.449 ns ns ns ns 0.285
SO2 ns 0.345 ns ns 0.301 ns ns 0.783 ns ns

4. Discussion
The present study presents the results of the use of two different
species e a lichen and a moss e as a passive biomonitors. In
transplantation studies, lichens and mosses taken from relatively
clean sites undergo substantial changes when exposed to more
polluted areas. Our study results were in accordance with such a
tendency.
Changes in the composition of photosynthetic pigments is found
to be the most sensitive parameter to assess stress of biomonitors
induced by a wide range of air pollutants (Boonpragob, 2002; Shukla
and Upreti, 2007; Bajpai et al., 2013a; Mateos and Gonzalez, 2016).

_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009
6 _ V. Galinyte_ / Atmospheric Pollution Research xxx (2016) 1e8
G. Sujetoviene,
In the present study, content of chlorophyll a and b in lichen and
moss was signicantly negatively affected at the urban environ-
ment. The samples of the lichen transplanted to the sites with
highest concentrations of the pollutants (urban and residential)
indicated highest decrease in pigment content. This phenomenon
was also observed in other studies where the content of photosyn-
thetic pigments decreased as the effect of the urban environmental
conditions (Can ~ as and Pignata, 1998; Majumder et al., 2013).
Transplants of the same lichen species (E. prunastri) from urban
sampling sites also displayed a decrease in photosynthetic pigments
along with a greater proportion of nitrophilous species (Lackovicova 
et al., 2013). Similar response of photosynthetic changes in the
lichen Everniastrum neocirrhatum was also found by Zambrano et al.
(1999) transplanted near Mexico city where notably loss of chloro-
phyll b was observed at the urban site. The gaseous compounds in
the air along with the accumulation of pollutants such as heavy
metals also can disturb the photosynthetic apparatus in urban and
industrial areas. Despite the level of air pollutants was not relatively
high during the exposure of the present study chlorophyll degra-
dation could be as a result of oxidation caused by air pollutants (Cuny
et al., 2002). A linear relationship between NOx and SO2 concen-
trations and the pigment content conrmed the negative impact of
these gaseous air pollutants. The fertilizing inuence of the air
pollutants as nutrients on the pigment content was not in accor-
dance with results of other studies (Ra et al., 2005; Satya and Upreti,
2009; Sujetoviene and Sliumpaite, 2013) where higher chlorophyll
content was associated with enhanced deposition sites.
The analysis of chlorophyll a uorescence parameters is
considered as an important approach for evaluating the health or
integrity of the internal apparatus during the photosynthetic pro-
cess (Tretiach et al., 2007b; Piccotto et al., 2011). Chlorophyll uo-
rescence has been used more intensively in recent studies on the
vitality of transplanted lichens and mosses (Tuba et al., 1997;
Fernandez-Salegui et al., 2006; Tretiach et al., 2007b; Piccotto
et al., 2011; Paoli et al., 2011). The present study shows that the
exposure of the samples to urban pollutants caused a signicant
decrease in the Fv/Fm of samples transplanted in the urban and
residential sites. In the case of the moss, uorescence decreased
more than that in lichen at the urban and residential sites what
indicated that the moss suffered the exposure conditions more. This
is in accordance with study from Tretiach et al. (2007a) which re-
sults showed that Fv/Fm was affected more in the moss H. cupressi-
forme than in the lichen P. furfuracea. The low values of Fv/Fm were in
accordance with the data relating the decrease in the chlorophyll
content in the lichens exposed to pollutants under these conditions.
Some eld experiments were conducted to investigate whether
atmospheric pollutants in the urban environment inuence the
photosynthesis performance of lichens and suggested that the
disintegration of the photobiont chlorophyll was caused by air
pollution (Niewiadomska et al., 1998; Garty et al., 2001; Piccotto
et al., 2011).
Lipids are major cell membrane components, essential for
various biological functions including cell growth and division of
normal tissues. Malondialdehyde is a subproduct (MDA) of lipid
peroxidation generated by free radicals and various gases (nitrogen
oxides, ozone, sulphur oxides). Its presence indicates damage to
cellular membranes. Therefore, the amount of MDA formed as a
result of oxidation processes is usually used as a parameter which
indicates the damage induced by oxidant atmospheric pollutants
on lichens (Can ~ as et al., 1997; Gonza lez and Pignata, 1997;
Majumder et al., 2013). In our study, a prolonged transplantation
period in the polluted environment induced oxidative stress in the
lichens. The observed increase in MDA concentration in the lichens
_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009
transplanted in the urban area is in accordance with the results of
previous studies (Can ~ as et al., 1997; Majumder et al., 2013). The
highest concentrations of MDA were found in the heavy transport
and industry sites (Gonza lez et al., 1996). However, an opposite
trend was characteristic of the mosses. The moss samples trans-
planted in the urban environment experienced stress at the
beginning of the transplantation and MDA content increased as an
indicator of damage by oxidant pollutants. After 2-months, the
MDA content remained higher in the urban environmental sites as
compared to the control site, but the difference was not signicant.
This indicated that the moss was able to cope with the environ-
mental stress.
The cell membrane damage, expressed by changes in electrical
conductivity, is considered as one of the most sensitive and suitable
indicators of the impact of air pollution stress on the vitality of li-
chens and mosses (Tarhanen et al., 1996; Garty et al., 2000; Paoli
and Loppi, 2008; Munzi et al., 2009). The results of the present
study indicated that the increase in electrolyte leakage as a result of
the destruction of the cell membrane in the lichen E. prunastri and
moss P. crista-castrensis was observed in the urban environment.
This means that in the presence of pollutants, the cell membrane of
the transplants experienced damage since it is the rst site of
biological interaction with toxic substances.
Increased membrane lipid peroxidation in the transplanted li-
chens and mosses along with enhanced electrolyte leakage are in
accordance with the results of the previous studies (Can ~ as et al.,
1997; Carreras et al., 2009; Sujetoviene and Sliumpaite, 2013)
which suggest that at a high concentration elements become toxic
by reducing membrane integrity. The most probable explanation
for the induced damage by oxidative processes was proposed by
Tretiach et al. (2007b) e NO2 oxidizes membrane phospholipids
forming free radicals and increases the formation of reactive oxy-
gen species (ROS) while NO reactions with hydrocarbons and ROS
form many toxic compounds. The observed correlation with ni-
trogen oxides could be the reason for the changes of physiological
parameters. Data of our study showed induced oxidative stress and
cell membrane damage as deleterious effects of air pollution at
urban environment.
The difference between the two species could be related to the
different ecological requirements. E. prunastri is light demanding
lichen occupying relatively warm and dry habitats according to
the species indicator values (Wirth, 1991). This implies that it is
able to survive and is resistant to prolonged drought and high
light intensity. On the contrary, P. crista-castrensis prefers a
moister environment and is tolerant to more shady conditions
(Dll, 1991). The analysis showed that the exposure in bags was
not the reason for the induction of physiological stress in its
natural growing environment. The exposure of the transplanted
material in the city was a substantial change of environmental
conditions related to the climate and air quality during the
exposure period.
The results of this study indicated that presented levels of pol-
lutants induced deleterious effects on the transplants. The exposure
of both organisms in the urban environments resulted in the
reduction of sample vitality. Both species showed no resilience
because all the measured parameters decreased or indicated
damage after the exposure. Comparing the two species, the moss
rather than lichen experienced more damage regarding chlorophyll
a content, uorescence reduction and higher cell membrane
damages in the urban environment. Results of the present study
indicated induced oxidative stress and cell membrane damage in
the transplants as deleterious effects of air pollution at urban
environment.
 _ V. Galinyte_ / Atmospheric Pollution Research xxx (2016) 1e8
G. Sujetoviene,
Conict of interest
There is no conict of interest.
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_ G., Galinyte,
Please cite this article in press as: Sujetoviene, _ V., Effects of the urban environmental conditions on the physiology of lichen and
moss, Atmospheric Pollution Research (2016), http://dx.doi.org/10.1016/j.apr.2016.02.009