Spatial and temporal variation of biomonitors conductivity at Lisbon city, Portugal

A.M.J.Cruz1, N. Canha2,*, M.C. Freitas2, S.M. Almeida2, H.Th. Wolterbeek3

1

Department of Science and Technology, Technology and Management High School of Oliveira do

Hospital, Coimbra Polytechnic Institute, Rua General Santos Costa, 3400-124 Oliveira do Hospital, Portugal
2 3

Instituto Tecnolgico e Nuclear, Instituto Superior Tcnico, Universidade Tcnica de Lisboa, Portugal

Department of Radiation, Radionuclides and Reactors, Faculty of Applied Sciences, Delft University of Technology, Mekelweg 15, 2629 JB Delft, The Netherlands *nunocanha@itn.pt

ABSTRACT A biomonitorization survey using transplated lichens from a clean area was done in the urban area of Lisbon, Portugal, at primary schools courtyards in 2008 and 2010. Two species of lichens, Parmotrema bangii and Parmelia caperata, were collected from clean areas (Azores and a clean rural zone in the center of Portugal, respectively). For each sampling period, the spatial variation of the electric conductivity results was determined to assess the city areas where the pollution had a greater impact on the lichens. Temporal evolution of the conductivity was analysed as well, concerning the two sampling periods, to understand if the pollution impact areas had changed.
Keywords: Biomonitors, Lichens, Conductivity, Spatial and Temporal variation, Lisbon, Air Quality

I.

INTRODUCTION

The potential of transplanted lichens as effective biomonitors of air quality have been emphasized by several studies in the past years (Sloof and Wolterbeek, 1991; Freitas et al., 1999). Lichens stress caused by pollution can be shown by physiological parameters which can supply relevant information on the pollution impact (Garty et al., 2001). Changes in membrane permeability to ions, characterized by the electric conductivity of the lichen samples, are one of the most sensitive physiological responses to environmental stress. In fact, the loss of electrolytes correlates with the presence of gaseous pollutants, catalysts of lipid membrane peroxidation and with some high heavy metal cellular concentrations (Paoli et al., 2011; Garty et al., 2002; Godinho et al., 2004). The ability of lichens to accumulate levels of elements in excess of physiological requirements in close correlation with atmospheric elemental levels has led to their wide-scale application as practical biomonitors of inorganic atmospheric contamination (Canha et al., 2010; Munzi et al., 2009). A significant reduction in total sampling costs and the possibility of an intensive assessment of a certain attribute in space and time increased the interest for this environmental monitoring tool (Canha et al., 2012; Cruz et al., 2011). A biomonitoring survey using transplanted lichens from a clean area was done in the urban area of Lisbon, Portugal, at primary schools courtyards in 2008 and 2010. The aim of this work was to assess the spatial and temporal distribution of the lichen electric conductivity in the whole city of Lisbon and to understand the influence of the meteorological conditions on it.

II. EXPERIMENTAL A. STUDY AREA

This study was performed in the municipality of Lisbon, Portugal, which has a resident population of 545245 inhabitants, according to the National Institute of Statistical of Portugal (Instituto Nacional de Estatstica, 2011). Lisbon is the largest city of Portugal and the westernmost capital in mainland Europe. Figure 1 shows the 22 primary schools of Lisbon where this study was conducted.

Figure 1. Spatial distribution of the primary schools in Lisbon (Portugal), where the transplanted lichens were exposed.

B.

Transplanted Lichens

This study was conducted in two different years: 2008 and 2010. For each phase, the transplanted lichens had different origins but both had the characteristics of clean air areas. In 2008, the Parmotrema bangii (Vain.) Hale lichen and tree bark (Cryptomeria japonica (L. fil.) D. Don) were collected in So Miguel, Azores islands (374725 N; 253812 W). Two collecting campaigns were done in January and May to correspond to the winter and summer exposure campaigns. The periods between sampling in Azores and exposure in Lisbon were 2 and 3 weeks, in winter and summer campaigns, respectively. In 2010, the Flavoparmelia caperata (Vain.) Hale lichen and tree bark (Olea europaea L., most known as olive tree) were collected in a rural area of Portugal mainland (390320 N, 81042 W), about 93 km away from Lisbon city. Two collecting campaigns were done in April and July for the winter and summer exposure campaigns. The period between sampling and exposure in Lisbon was, at maximum, 1 week.

C.

Transplant Method

At the laboratory, the collected lichen samples were set into tree bark pieces of around 6 cm x 6 cm each and, afterwards, they were hanged at the courtyards trees (at the tree branches at about 1.80m height) of the studied schools, using nylon strings. A total of 3-4 samples were exposed per schools to act as

replicates. Per each campaign, ten of these pieces were kept in the laboratory unexposed to Lisbon air pollution to estimate the baseline of the lichen samples (from the clean area where they were collected). No more than 1 week prior to the arrival to the laboratory, these samples were processed as explained below. Table 1 shows the details of the lichen exposure campaigns in 2008 and 2010. Although a set of 3-4 replicates were exposed in each school, after the exposure periods some of the samples (sometimes only some replicates but others all of them) were loss and, therefore, from the initial set of 22 primary schools only was possible to gather results to a lower number of schools. Table 1. Details of the lichen exposure campaigns in 2008 and 2010. Year 2008 Campaign Winter Summer Winter Summer Total of Studied Schools 14 20 14 8 Exposure Period January - May June - October April - June July - September Total of exposure days 120 120 69 43

2010

D.

Sample preparation and Electric Conductivity Measurement

After the exposure periods, lichen samples were sorted and cleared of extraneous material (dust, leaf debris, fungus contamination and degraded material) at the laboratory. For the conductivity measurements, the established procedure (Marques and Freitas, 2005) was followed: i) the lichen samples were rinsed 3 times for 5 seconds with double distilled water and air dried for 24 hours; ii) 100 mg of each sample was immersed in 10 mL for 60 minutes; iii) the electric conductivity was measured afterwards with an electric conductivity meter (712 Metrohm Conductometer). To the non-exposed samples, it was applied the same procedure.

E.

Meteorological Data

The meteorological data for the 4 exposure periods (temperature, humidity and wind directions) was collected was well (Russias Weather, 2011).

F.

Statistical Analysis

Statistical analyses were performed with Statistica and the geostatistical modelling maps were built using GeoMS software (GeoMS, 2011) with the aim of assessing the spatial distribution of the electric conductivity in the whole city of Lisbon.

III. RESULTS AND DISCUSSION Figure 2 shows the average values of the conductivity obtained for each exposure campaign, as well, the respective averages values of temperature and relative humidity. Applying a Mann-Whitney test to the 4 sets of conductivity, it was possible to conclude that all the studied binary groups are significantly different among themselves, since the p-value was below 0.05. A Kruskal-Wallis test resulted in a pvalue=0.0000, which means that the seasonal averages are significantly different between themselves.

Figure 2. Average values of the lichens electric conductivity, relative humidive and temperature for the 4 periods of lichens exposure: winter 2008, summer 2008, winter 2010 and summer 2010.

It is possible to assess that lower temperatures and relative humidities contribute for lower electric conductivities, which was expected since in these conditions the lichen is not under so much stress due to the meteorological conditions. In fact, the exposure periods in 2010 presented higher values of temperature and relative humidity which conducted to the lichen cell membrane damage and, therefore, to higher values of electric conductivity. The exposure campaign of summer 2010 had the shortest duration (43 days) and, although it registered the higher temperatures and lower relative humidities, the electric conductivity did not reflect it. This fact can be explained by the reduced exposure time, by which the lichens managed to keep their cell membrane integrity. Set of 3-4 subsamples of lichens were exposed in all the courtyards of the 22 primary schools. However, after the exposure period, several samples were lost. With the gathered data of electric conductivity from the collected lichen samples, it was possible to build the Lisbon spatial distribution maps of this parameter for the 4 different exposure periods with the GeoMS software. Figures 3, 4, 5 and 6 show the spatial distribution maps of electric conductivity for Lisbon city.

Figure 3. Spatial distribution of biomonitors conductivity in the exposure period of winter 2008 (total of samples collected after exposure: 14).

Figure 4. Spatial distribution of biomonitors conductivity in the exposure period of Summer 2008 (total of samples collected after exposure: 20).

Figure 5. Spatial distribution of biomonitors conductivity in the exposure period of winter 2010 (total of samples collected after exposure: 14).

Figure 6. Spatial distribution of biomonitors conductivity in the exposure period of summer 2010 (total of samples collected after exposure: 8).

Figure 6 show the spatial distribution map of the lichens conductivity built with data from only 8 schools. The geostatistical model of the summer 2010 data resulted in a map not very well defined probably due to small number data that was gathered. It is possible to distinguish a major area in the center of Lisbon with higher conductivities and no intermediary values but, overall, the final result is a blur. This fact confirms that data sets with a lower amount of samples, as 8 in case of summer 2010, are not adequate for application of GeoMS software. On the other hand, in the other 3 maps is possible to visualize areas of higher values of conductivity distributed in space. In fact, higher electric conductivities are usually nearby main roads and avenues of the city. The area nearby of the Avenida da Liberdade, which is one of the most polluted areas in Lisbon (Borrego, et al., 2000) due its intensive traffic and characteristic topography, presented higher conductivities in all the maps which confirms the high levels of pollution already assessed through other studies. In 2008, only two sites presented higher values of electric conductivity in both exposure periods. One was the one explained above and the other was at northwest and northeast for winter and summer periods, respectively. Figure 7 shows the wind directions during the exposure periods and it is possible to observe that the main wind directions in these exposure periods were opposite to second area of higher electric conductivity. In 2010, the winter period show that, beside the area described above, exists two other small areas in the southwest of Lisbon city with higher electric conductivities and, once again, they are nearby main highways (A5 and IP7 Eixo Norte-Sul). Both mentioned highways are radial connections and part of the structural road network of the city (CML, 2005), which serves to support over long distances. In fact, this structural road network includes more two main roads: CRIL and 2 Circular, which are circular connections to the city. These main roads are part of the 1st level of the structural road network which, along with the 2 nd, are 32% of the total network but absorb 72% of the total road traffic of the city (with maximums with nearly 60000 vehicles in transit in rush hours: 8am and 6pm). The primary schools which are located nearby these main roads present higher values of electric conductivity. Therefore, electric conductivity is, in fact, a good indicator of the main of focus of pollution.

Figure 7. Wind directions during the exposure periods (left 2008 and right 2010).

IV. CONCLUSIONS The spatial distribution of lichen conductivity at the end of the winter and summer campaigns for the study periods shows variability between years and across the studied area. Conductivity was significantly higher at the end of summer than at the end of winter, which is explained by the significantly lower humidity and higher temperatures in summer. The primary schools nearby the main roads with high intensity of road traffic present higher electric conductivities which confirm a higher degree of damage of the lichen membrane cells due to higher pollutants concentration. This physiological indicator of the lichen stress symptoms is a very effective tool of the biomonitoring studies to understand main focus of pollution and changes over the time.

V. ACKNOWLEDGMENTS Financial support by Fundao para Cincia e Tecnologia (FCT; Portugal) through research contract PTDC/SAU-ESA/65597/2006 is gratefully acknowledged. N. Canha thanks FCT for his PhD grant (SFRH/BD/72272/2010) and S.M. Almeida thanks FCT for Cincia 2007 support.

VI. REFERENCES AND FOOTNOTES Borrego, C., Tchepel, O., Barros, N., Miranda, A.I., 2000. Impact of road traffic emissions on air quality of the Lisbon region. Atmospheric Environment 34(27), 4683 - 4690. Canha , N., Freitas , M.C., Almeida, S.M., Pacheco, A.M.G., 2010 . Element-enrichment factors in Parmotrema bangii and Cryptomeria japonica of Portuguese islands of the central North Atlantic. International Journal of Environment and Health 4(2/3), 120136. Canha, N., Almeida-Silva, M., Freitas, M.C., Almeida, S.M., Wolterbeek, H.T., 2012. Lichens as biomonitors at indoor environments of primary schools. Journal of Radioanalytical and Nuclear Chemistry 291(1), 83-88. CML, 2005. Lisboa: o desafio da mobilidade, CML, Cmara Municipal de Lisboa, 2005, 294 pp.. Cruz, A.M.J., Freitas, M.C., Verburg, T.G., Canha, N., Wolterbeek, H.Th., 2011. Response of exposed bark and exposed lichen to an urban area. Proceedings in Radiochemistry 1, 363369. Freitas, M.C., Reis, M.A., Alves, L.C., Wolterbeek, H.Th., 1999. Distribution in Portugal of some pollutants in the lichen Parmelia sulcata. Environental Pollution 106, 229235. Garty, J., Weissman, L.,Cohen, Y., Karnieli, A., Orlovsky, L., 2001. Transplanted lichens in and around the Mount Carmel National Park and the Haifa Bay industrial region in Israel: physiological and chemical responses. Environmental Research 85(2), 159-176. Garty, J., Tamir, O., Cohen, Y., Lehr, H., Goren, A.I., 2002. Changes in the potential quantum yield of photo system II and the integrity of cell membranes relative to the elemental content of the epilithic desert lichen Ramalina maciformis. Environmental Toxicology and Chemistry 21(4), 848858. GeoMS software, 2011. http://cmrp.ist.utl.pt/index.php?lg=1&cont=9, access: 10 June 2011. Godinho, R.M., M.C. Freitas, e H.Th. Wolterbeek. Assessment of lichen vitality during a transplantation experiment to a polluted site. Journal of Atmospheric Chemistry, 2004: 355-361.

Instituto Nacional de Estatstica, 2011. http://censos.ine.pt., accesso:4 November 2011. Marques, A.P., Freitas, M.C., 2005. Cell-membrane damage and element leaching in transplanted Parmelia sulcata lichen related to ambient SO2, temperature, using transplants. Environmental Science & Technology 39(8), 2624-2630. Munzi, S., Pisani, T., Loppi, S., 2009. The integrity of lichen cell membrane as a suitable parameter for monitoring biological effects of acute nitrogen pollution. Ecotoxicology and Environmental Safety 72, 20092012. Paoli, L., Pisani, T., Guttov, A., Sardella, G., Loppi, S., 2011. Physiological and Chemical response of lichens transplanted in and around an industrial area of south Italy: Relationship with the lichen diversity. Ecotoxicology and Environmental Safety 74, 650-657. Russias Weather, 2011. http://meteo.infospace.ru, access: 27 October 2011. Sloof, J. E., Wolterbeek, H.Th., 1991. A national monitoring survey using epiphytic lichens as biomonitors of trace-element air pollution, Lichenologist 23, 139165. BIOGRAPHIES Ana Janurio Cruz She is PhD student in Environmental Science, in Delft University of Technology, Delft, Netherlands. Her research interest is to characterize the air quality in Portugal, with a special focus in Lisbon, and to estimate the health risks associated with exposure to atmospheric pollution. She finished a biomonitoring survey of Lisbon foreseen aiming to obtain a spatial screening of atmospheric pollution. Her works are inserted in the scientific activities of the Neutron Activation in Environment, Nutrition and Epidemiology group, at the Unity of Reactors and Nuclear Safety of the Technological and Nuclear Institute. Prof. Cruz is lecturer at the College of Technology and Management of Oliveira do Hospital (Polytechnic Institute of Coimbra, Portugal), where she is also the coordinator of the Technological Specialization Course in Quality Management and the Environment. Nuno Canha He is a Junior Researcher in the project for indoor-air studies at Lisbon (Portugal) elementary schools, which is part of the scientific activities by the Neutron Activation in Environment, Nutrition and Epidemiology group of the Instituto Tecnolgico e Nuclear, Portugal. He holds a MSc in chemistry from the Technical University of Lisbon, Portugal. He is primarily involved in the determination of chemical elements and natural radionuclides in filter samples through instrumental neutron activation analysis and gamma spectrometry. He is a PhD student in Environmental Science, in Delft University of Technology, Delft, The Netherlands. His current work deals with assessment of bioaerossols (fungi, bacteria and pollens) and total particulate matter (TPM) inside school classrooms, biomonitoring studies with lichens of indoor and outdoor environments of the primary schools and, as well, bioremediation of contaminated mining areas in Portugal. Maria do Carmo Freitas She is a Researcher in the Unity of Reactors and Nuclear Safety at the Technological and Nuclear Institute and colaborator of the Neutron Activation in Environment, Nutrition and Epidemiology scientific activity. She is primarily involved in the determination of chemical elements in environmental and nutritional samples using nuclear analytical methods. She has many years of

experience in analyzing air samples and biomonitor samples, either at urban or rural areas. Source apportionment for characterizing local, regional and remote sources has been done with the results on airborne particles and the results on biomonitors. Her current research interests include low-level counting of natural radioactivity, Compton suppression gamma-ray spectrometry, prompt-gamma radiation. Speciation is nutritional and environmental samples is also one of her interests. Her interest in environment also concerns the potential association to epidemiological risks. Susana Marta Almeida She holds a degree in Environmental Engineering (1998, Universidade Nova de Lisboa) and a PhD in Environmental Sciences (2004, Aveiro University). Since 1998 she has been developing research in Environment Sciences in several Portuguese research institutions Technological and Nuclear Institute (ITN), Aveiro University (UA), National Institute of Engineering, Technology and Innovation (INETI) and Welding and Quality Institute (ISQ). She has participated and coordenated several national and European R&D projects and is author and co-author of more than 40 scientific publications on atmospheric pollution, indoor air quality, epidemiology and risk assessment. She is the Coordinator of the Neutron Activation in Environment, Nutrition and Epidemiology scientific activity in ITN. Hubert Wolterbeek He has been working in the Department of Radiochemistry of the Interuniversity Reactor Institute, Delft, and is now Head of the research section Radiation and Isotopes for Health (RIH), Delft University of Technology. He graduated in Physiology from Utrecht University, the Netherlands. His focus of research has been on the development and use of radioanalytical and radiotracer techniques in issues such as biomonitoring of metal air pollution, plant and animal physiology, environmental and general chemistry, with progressively growing attention for implication on human health.