UNIVERSITY OF PANNONIA DOCTORAL SCHOOL OF CHEMICAL AND ENVIRONMENTAL SCIENCE Use of siliceous algae in the environmental reconstruction in the Carpathian Region Ph.D. theses Éva Soróczki-Pintér Supervisors: Prof. Dr. Judit Padisák Head of Department Professor, Doctor of the Hungarian Academy of Sciences Department of Limnology, Institute of Environmental Sciences University of Pannonia dr. Krisztina Buczkó museologist Hungarian Natural History Museum Department of Botany Veszprém 2015 Use of siliceous algae in the environmental reconstruction in the Carpathian Region Ph.D. theses Éva Soróczki-Pintér Supervisors: Prof. Dr. Judit Padisák Head of Department Professor, Doctor of the Hungarian Academy of Sciences Department of Limnology, Institute of Environmental Sciences University of Pannonia dr. Krisztina Buczkó museologist Hungarian Natural History Museum Department of Botany 2 1. BACKGROUND AND INTRODUCTION In the recent decades palaeolimnological researches have been achieving an increasing role in the past climate change studies. The long-time history of lake can be reconstructed by revealing the physical, chemical and biological processes in lake sediment using biotic and abiotic proxies. The arctic and alpine lake sediments, which accumulate slowly over time and can be well determined, are ideal for palaeoecological studies. Especially being aware of the alpine habitats are considered one of the most vulnerable areas of the biosphere regarding the climate change (e.g. Tinner & Kaltenrieder 2005). The entire lake history back to thousands of years including hydrology, in-lake productivity and vegetation development on catchment area can be reconstructed using successional pattern of biological microfossils (e.g. Davidson & Jeppesen 2013). Diatoms are one of the most commonly used biotic groups (called proxy) of palaeoenvironmental studies focusing on environmental- and the climate change. Diatoms are clear favored taxonomic group, ecological optima and tolerance values of the species are known. Silicon diatom frustules are well preserved for a long time in the sediments. Diatom preparations are durable and can be stored in a small space (Charles et al. 1994). Additionally, diatom community is one of the five indicator organism groups (phytoplankton, phytobenthos, macrophytes, macrozoobenthos, fish) and the most important element of the phytobenthos used for biological water qualification in surface waters (Water Framework Directive Implementation in Hungary, 2000). Chrysophytes are known as a diverse group of freshwater algae consisting of over 1000 described species (Duff et al. 1995). They are most common in nutrient-poor, acidic, oligotrophic waters (Duff et al. 1995). By mixotrophic nutritional strategy (Sandgren 1988) chrysophytes able to adapt to extreme physical or chemical conditions (e.g. Zeeb & Smol 1995). All chrysophytes during unfavorable environmental conditions can produce siliceous resting stages, called stomatocysts. The cysts are often well preserved in the sediments of most lakes due to resistance to dissolution. Currently more than 800 morphotypes are known according to literature and database (Duff et al. 1995, Facher & Schmidt 1996, Wilkinson 2001, Pla 2001, Kamenik et al. 2001, Kamenik 2010). In recent decades the cysts have high potential in palaeoecological studies. Some cysts are characterized by well-defined ecological optimum values showing a tight tolerance to physical (e.g. temperature, ice cover periods) and chemical (e.g. pH, salinity, conductivity) variables (Dixit et al. 1989, Pla & Anderson 2005). The cysts have been used to reconstruct winter and early spring temperature (e.g. De Jong & Kamenik 2011) and unique ice cover length, lake mixing and stratification patterns triggered by seasonality (e.g. Pla-Rabes & Catalan, 2011). Additionally, in recent decades studies focusing on taxonomic and morphological cyst characteristics have been carried out intensively in arctic, subarctic (e.g. Pla & Anderson 2005) and temperate mountain lakes (e.g. Lotter et al. 2002, Schmidt et al. 2006, Facher & Schmidt 1996, Pla 2001) as well as large lowland lakes (Firsova et al. 2012). The knowledge on geographical distribution of stomatocysts has generally been scarce, but in Carpates cyst taxonomy has been relatively well documented (e.g. Péterfi 1967, Cabała & Piàtek 2004, Cărăuş). Palaeoenvironmental studies in the Central and Eastern European region, including Southern Carpathians, are poor (Lotter & Hofmann 2003). To overcome this lack in 2007 leading dr. Enikő Magyari a Late Quaternary complex, multi-proxy study started using palaeoecological and palaeolimnological methods in four glacial lakes (Lake Brazi, Lake Gales, Lake Bukura and Lake Lia) of Retezat Mountains in the Southern Carpathians to understand the Late-glacial and Holocene environmental changes. 3 In this study, as part of a Late Quaternary multi-proxy project my research focused on siliceous algae microfossils (diatoms and Chrysophycean cysts) analysis from Lake Gales. This research using high resolution diatom and low resolution cyst analyses was focused on ecosystem reponses during late-glacial and Holocene periods which were triggered by climatic changes. The high resolution diatom analysis and inferred lake-level reconstruction, additionally preliminary cyst-database are niche studies for the Southern Carpathians. 2. MAIN OBJECTIVES (1) The primary goal of my research was to determine the fossil diatom taxa in sediment of Lake Gales situated on the northern slope of the Retezat Mountains. (2) Based on the previous point, my goal was to describe successional development of diatom flora of Lake Gales regarding to late-glacial and Holocene periods. (1) Further goal was to reconstruct the lake water level changes in Lake Gales based on the dominant diatom taxa and changes in lifeform. The questions to be answered were: i. Can you detect the changes or increases in water level in ontogeny of Lake Gales? ii. What is the main factor that determines appearance or dominance/absence of the diatom taxa? iii. Is it possible to infer an environmental/climatic event which triggered the change in diatom composition? iv. Is it possible to infer any regional or global scale climatic event/anomaly? v. If so, is there a regional or global level in time, or delayed climate change having similar amplitude and similar response of biotic proxy? (3) My research was extended to analyse the chrysophycean cysts having silica skeleton; my goal was to determine the cyst:diatom frustules ratio (C:D) in every sample of the core to estimate in-lake trophity approximately. (4) Further goal was to estimate the distribution of the cysts and to describe the dominant morphotypes throughout the core by low-resolution cyst analysis. The questions were: i. Is it possible to discover typical cyst morphotypes in the late-glacial and the Holocene period? ii. Can a typical cyst morphytype(s) appear parallel with the dominant diatom in high lake level of the lake history? 3. MATERIALS AND METHODS Sampling site The Retezat Mts. are one of the wettest alpine regions of the Romanian Carpathians because of the combined effect of Mediterranean and oceanic influences. Along the peaks the temperature is below 0 °C most of the year, in the summer months sometimes occurs snowfall (Jancsik 2009). After retreat of the glacials formed in late-glacial period various glacial lakes were formed in the subalpine and alpine belts, and their basins (Urdea, 1993, 2000; Vespremeanu-Stroe et al., 2008). 4 The studied Lake Gales (Lacul Galeş) is located on the northern slope in the Retezat Mountains (Southern Carpathians) at 1,990 m a.s.l., 150 meters above the timberline zone consisting spruce (Pinus abies) and stone pine (Pinus cembra), at the bottom of Gales glacialvalley. Lake Gales is the third deepest lake of the Retezat Mts., the maximum water depth is 20.5 meters and the surface area is 3,68 ha. The lake is fed with a stream inlet on the south side and in the opposite side Stream Gales flows across the entire valley. Methods The sediment core from Lake Gales was obtained with a modified Kullenberger piston corer in the deepest part of the lake at a water depth of 19.5 m in August 2007. The 328 cm sediment core was taken in one drive and was stored at 2 oC in the lab until further treatment. The analyses of the following components were carried out: macrofossils-, stomata-, pollen-, siliceous algae (diatom and chrysophycean cysts), major and trace elements, organic matter, micro-, and macro charcoal components and then in 2013 biogenic silica content was determined. A chronological framework of sediments Gales-3 was established using 9 AMS 14 C age determinations: on four terrestrial plant macrofossils and five Cladocera remains (mainly eggs) were analyzed. Four tie-points obtained by comparing the pollen spectra of Gales-3 with Lake Brazi (TDB-1) were also used for age-depth modelling in late-glacial (for details see Magyari et al. 2012). The results of the radiocarbon dating, age-depth modelling, sediment stratigraphy, organic content measurements (LOI), sediment accumulation rates (SAR) in Lake Gales were described in Magyari et al. (2009b, 2012). The digestion of siliceous algae (chrysophycean cysts and diatoms) followed standard methods (Battarbee 1986). For siliceous algae analyses samples were taken from every 4 cm. After the acid digestion by HCl and H2O2, the aliquot suspensions were evaporated and embedded in Zrax (R.I=1.7) or Pleurax (R.I=1.73). At least 300 valves were counted in the samples using a light microscope (LEICA DM LB2 with 100x HCX Plan Apo objective), converted to relative frequency percentages. To reduce the taxonomic uncertainties scanning electron microscope (Hitachi S-2600N) was used. Taxonomy follows Krammer and LangeBertalot (1986-1991), Lange-Bertalot & Krammer 1989, Lange-Bertalot 2001, Lange-Bertalot és Metzeltin 1996, Spaulding et al. 2010, Lowe et al. 2014, Hamilton et al. 2014 and we used the following on-line taxonomy web site, http://westerndiatoms.colorado.edu of Institute of Arctic and Alpine Research (USA). We controllod the current nomenclature of diatom taxa ont he web page http://www.algaebase.org (Guiry & Guiry 2014). The counting of Chrysophyta cysts occurred at the same time with counting of diatom frustuls per sample. Total chrysophycean stomatocysts (C) were expressed as a ratio with respect to the number of counted diatom frustules(D), the C:D ratio informs the in-lake productivity. The study of cyst flora was studied along the high-resolution diatom zones (DAZ), in selected samples per zone 200 cysts were counted and used SEM and where possible LM for ensuring taxonomic consistency and accuracy. The low resolution cystdatabase can be applied for comparison of other Retezat lake’s flora. New cysts with „C” prefix referring to the „Carpathians” were described following the International Statospore Working Group (ISWG) guidelines (Cronberg & Sandgren 1986). Cyst taxonomy follows Duff et al. (1995), Facher & Schmidt (1996), Wilkinson et al. (2001) and Pla (2001) and „Stom@ocyst”-called online cyst database by Kamenik (2010). The converted relative frequencies of diatom frustules were illustrated using Psimpoll 3.00 program (Bennett 2005) to determine diatom assemblages zones (DAZ) of Gales-core. For stratigraphical analysis was prepared on different diatom dataset to achieve the most accurate information of DAZ zones. The validation of varied databased diatom zones was carried out with pvclust package in R program (R Development Core Team). By Principal 5 ComponentAnalysis (PCA) in prcomp package (R program) we defined the separation of fossil diatom community using different diatom dataset. 4. THESIS POINTS (1) Diatom assemblages in Lake Gales A high resolution diatom analysis was carried out on the 328 cm long lacustrine sediment core in Lake Gales (45°23'6"N, 22°54'33"E). I distinguished 178 diatom taxa sampled whole sediment core. Two diatom species, Pinnularia julma Krammer & Lange-Bertalot and Navicula detenta Hustedt proved to be new data for algal flora of Romania. (2) Diatom succession in the sediment of Lake Gales, window to lake level changes Diatom-based lake level reconstruction at the first phase of lake onthogeny (Bølling/Allerød) referred to a shallow but permanent lake with increasing lake level. At the onset of cold and dry Younger Dryas (~12,900 cal. yr BP) I detected a sharp change in siliceous algae record, the siliceous remains were scarce and aerophytic taxa became dominant inferring seasonal aridity or dessication of the lake due to the sudden climatic deterioration. Between 12,500-11,700 cal yr BP, in the second part of YD benthic and periphytic diatoms were dominant suggesting a minor lake level increase and shallow lake conditions. From 11,700 cal yr BP likely coinciding with the onset of Holocene I detected a rapid diversified siliceous algae assemblages. Aquatic and terrestrial vegetation productivity (organic material content) sharply increased inferring, warm climatic conditions. The most remarkable changes in diatom record have been detected between 9500-9000 cal yr BP, when a spine like, lightly silicified diatom, Fragilaria gracilis Østrup became dominant having 70% relative abundance at 9210 cal yr BP. Fragilaria gracilis-peak was a unique, peculiar and well-dated event in diatom record inferring high lake level during the onthogeny of Lake Gales. The special diatom composition, minimum of LOI inferred organic content and decrease of C:D ratio suggested increasing erosion activity and decreasing in-lake productivity. The decrease of in-lake-productivity can be linked to the 9.2-ka widespread and significant climatic anomaly triggered by a meltwater pulse (MWP) into the North Atlantic resulted in the slowdown or shutdown of the thermohaline circulation (THC) at high - and midlatitudes on the Northern Hemisphere. After 9150 cal yr BP a shift started, the relative frequencies of Aulacoseira species increased gradually replaced the fragilaroid diatoms meaning high lake level but from ~5500 kal. yr BP periphytic, benthic taxa were prominent elements in diverse diatom profil. After ~3,700 cal. yr BP a sharp and determinative change was detected in diatom record: (tycho)planktonic Aulacoseira taxa became dominant elements in record replaced previously dominant fragilaroid taxa indicating high lake level and in-lake productivity. (3) Chrysophycean stomatocysts in Lake Gales (Southern Carpathian, Romania) Lake Gales was characterized by 83 cyst morphotypes knowing as cold tolerant, generally in slightly acidic and oligotrophic habitat-prefering types. I determined that the cyst record with various morphology in Lake Gales was very abundant. The flora showed some differences between Late-glacial and Holocene record. In the Lateglacial section the unornamented and 6 larger cyst forms were typical forms, the flora was diverse in early Holocene, while in the younger part of core ornamented forms became more abundant, but the small, simple forms were dominant in every sample of the core with the exception of the sample ~ at 9200 cal yr BP. I determined an abrupt shift in cyst record ~ at 9200 cal yr BP, when the density of C072 form showed a sharp peak. Following the International Statospore Working Group (ISWG) guidelines (Cronberg & Sandgren 1986) I described and documented seven new cyst morphotypes such as C500, C501, C502, C503, C504, C505, C506. The new cysts are enumerated from ”500” and “C” prefix referring to the “Carpathians”. 5. REFERENCES Battarbee, R.W. (1986) Diatom analysis. In: Berglund, B.E. (ed.): Handbook of Holocene Palaeoecology and Palaeohydrology. John Wiley & Sons, Chichester, New York, Brisbane, Toronto, Singapore. pp. 527-570. Bennett KD (2005) Psimpoll manual. http://www.kv.geo.uu.se/psimpoll.html. Accessed 12 Feb 2008 Cabała, J. & Piàtek, M. (2004) Chrysophycean stomatocysts from the Staw Toporowy Nizni lake (Tatra National Park, Poland). Annales de Limnologie 40: 149-165. Cărăuş, I. (2012) Algae of Romania - a distributional checklist of acutal algae. 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Lotter, A.F., Appleby, P.G., Bindler, R., Dearing, J.A., Grytnes, J˗A., Hofmann, W., Kamenik, C., Lami, A., Livingstone, D.M., Ohlendorf, C., Rose, N. & Sturm, M. (2002) The sediment record of the past 200 years in a Swiss high˗alpine lake: Hagelseewli (2339 m a.s.l.). Journal of Paleolimnology 28: 111-127. http://dx.doi.org/10.1023/A:1020328119961 Lowe, R.L., Kociolek, P., Johansen, J.R., Van de Vijver, B., Lange-Bertalot, H. & Kopalová, K. (2014). Humidophila gen.nov., a new genus for a group of diatoms (Bacillariophyta) formerly within the genus Diadesmis: species from Hawai'i, including one new species. Diatom Research 29(4): 351-360 Magyari, E.K., Braun, M., Buczkó, K., Hubay, K. & Bálint, M. (2009b) Radiocarbon chronology of glacial lake sediments in the Retezat Mts (S Carpathians, Romania): a window to Lateglacial and Holocene climatic and palaeoenvironmental changes. Central European Geology 52: 225-248. Magyari, E.K., Buczkó, K., Jakab, G., Braun, M., Pál, Z., Karátson, D. & Papp, I. (2009a) Palaeolimnology of the last crater lake in the Eastern Carpathian Mountains: a multiproxy study of Holocene hydrological changes. Hydrobiologia 631: 29-63. http://dx.doi.org/10.1007/s10750-009-9801-1 Magyari, E.K., Jakab, G., Bálint, M., Kern, Z., Buczkó, K. & Braun, M. (2012) Rapid vegetation response to lateglacial and early Holocene climatic fluctuation in the South Carpathian Mountains (Romania). Quaternary Science Reviews 35: 116-130. http://dx.doi.org/10.1016/j.quascirev.2012.01.006 Péterfi, L.Şt. (1967) Studies on the Rumanian Chrysophyceae. I. Nova Hedwigia 13: 117-137. Pla, S. & Anderson, N.J. (2005) Environmental factors correlated with chrysophyte cyst assemblages in low arctic lakes of south-west Greenland. Journal of Phycology 41: 957974. Pla, S. (2001) Chrysophycean cysts from the Pyrenees. Bibliotheca Phycologica, Band 109. Cramer, Berlin, Germany. 179 pp. 8 Pla˗Rabes, S. & Catalan, J. (2011) Deciphering chrysophyte responses to climate seasonality. Journal of Paleolimnology 46: 139-150. http://dx.doi.org/10.1007/s10933-011-9529-6 R Development Core Team (2012) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0, URL http://www.R-project.org/ Sandgren, C. D. (1988) Growth and reproductive strategies of freshwater phytoplankton. Cambridge University Press, Cambridge. 442 pp. Sandgren, C.D. (1991) Chrysophyte reproduction and resting cysts: a paleolimnologist’s primer*. Journal of Paleolimnology 5: 1-9. Schmidt, R., Kamenik, C., Tessadri, R. & Koinig, K.A. (2006) Climatic changes from 12,000 to 4,000 years ago in the Austrian Central Alps tracked by sedimentological and biological proxies of a lake sediment core. Journal of Paleolimnology 35: 491-505. Spaulding, S. A., Lubinski, D. J. & Potapova, M. (2010) Diatoms of the United States. l., http://westerndiatoms.colorado.edu Tinner W & Kaltenrieder P (2005) Rapid responses of high-mountain vegetation to early Holocene environmental changes in the Swiss Alps. Journal of Ecology 93: 936-947. Víz KeretIrányelv (2000) Az Európai Parlament és a Tanács 2000/60/EK irányelve (2000. október 23.) a vízpolitika terén a közösségi fellépés kereteinek meghatározásáról. Wilkinson, A.N., Zeeb, B. & Smol, J.P. (2001) Atlas of Chrysophycean Cysts, Volume II. Kluwer Academic Publishers, Dordrecht. 180 pp. Zeeb, B.A. & Smol, J.P. (1995) A weighted-averaging regression and calibration model for inferring lakewater salinity using chrysophycean stomatocysts from lakes in western Canada. International Journal of Salt Lake Research 4: 1-23. DOI: 10.1007/BF01992411 9 6. SCIENTIFIC ACTIVITY PAPERS RELATED TO THE TOPICS OF THE THESES Padisák, J., É. Soróczki-Pintér & Zs. Rezner (2003) Sinking properties of some phytoplankton shapes and relation of form resistance to morphological diversity of plankton an experimental study. Hydrobiologia 500: 243-257. K. Buczkó, E. Magyari, É. Soróczki-Pintér, K. Hubay, M. Braun M. Bálint (2009) Diatombased evidence for abrupt climate changes during the Lateglacial in the Southern-Carpathian Mountains. Central European Geology 52: 249-268. DOI: 10.1556 Soróczki-Pintér É., Buczkó K., Braun M., Magyari E. (2012) Késő-glaciális és Holocén vízszintváltozások a Retyezátban egy gleccsertó kovaalga összetétele alapján. Hidrológiai Közlöny 92: 64-67. Soróczki˗Pintér, É., Pla˗Rabes, S., Magyari, E.K., Stenger-Kovács, Cs. & Buczkó, K. (2014) Late Quaternary Chrysophycean stomatocysts in a Southern Carpathian mountain lake, including the description of new forms (Romania). Phytotaxa 170: 169-186. http://dx.doi.org/10.11646/phytotaxa.170.3.3 ABSTRACTS RELATED TO THE TOPICS OF THE THESES Buczkó, K., Soróczki-Pintér, É., Braun, M., Hubay, K., Magyari, E.K. (2011) Az elmúlt 15 ezer év környezeti változásainak lenyomata két retyezáti gleccsertó kovaalgáin. In: Bosnakoff M., Dulai, F. & Pálfy J. (eds): Program, előadáskivonatok, kirándulásvezető 14. Magyar Őslénytani Vándorgyűlés. Magyarhoni Földtani Társulat. Szeged, 2011. június 2-4. pp. 11-13 Buczkó, K., Magyari, E.K., Soróczki-Pintér, É., Braun, M., Hubay, K., Tóth, M., Korponai, J.(2011) Response to cooling: Late-glacial and Holocene changes of diatom assemblages and lake acidity in mountain and lowland lakes in the Carpatian Basin. In: Mindrescu, M., Veres, D. & Onac B.P. (eds) Climate change in the Carpathian-Balkan region during late Pleistocene and Holocene. Suseava, Romania, 9-12 June 2011. pp: 39-40. Soróczki-Pintér, É., Magyari, E.K. & Buczkó, K. (2013) Preuve fondée sur les algues siliceuses de l’augmentation du niveau d’eau et du refroidissement à court terme autour de 9.2-ka dans les Carpates du Sud, Roumanie. (Siliceous algae based evidence for short-term lake level increase and cooling around 9.2-ka BP in the South Carpathian Mountains, Romania). In.: Rimet, F., Bouchez, A., Ector, L. & Montuelle, B. (eds.): INRA Science and Impact, 7th Central European Diatom Meeting (CE-Diatom) - 32nd meeting of the FrenchSpeaking Diatomists Association (ADLaF), Thonon-les-Bains, France, 16-20 sept. 2013. ISSN 978-2-7466-6166-0. pp. 77 -80. PRESENTATIONS RELATED TO THE TOPICS OF THE THESES Soróczki-Pintér É. (2010) Kovaalgák jelentősége a paleolimnológiai állapotrekonstrukciókban. „Aktuális eredmények a kriptogám növények kutatásában” konferencia. Eger, 2010. május 21. Soróczki-Pintér, É., Magyari, E. & Buczkó, K. (2011) Assessment of diatoms as indicators of late-glacial and Holocene climatic change in glacial lakes in the Southern-Carpathian Mountains. „5th CeDiatom Meeting. Szcecin, Poland, 24-27 March 2011. 52. pp. Buczkó, K., Soróczki-Pintér, É., Braun, M., Hubay, K., Magyari, E.K. (2011) Az elmúlt 15 ezer év környezeti változásainak lenyomata két retyezáti gleccsertó kovaalgáin. In: Bosnakoff 10 M., Dulai, F. & Pálfy J. (eds): Program, előadáskivonatok, kirándulásvezető 14. Magyar Őslénytani Vándorgyűlés. Magyarhoni Földtani Társulat. Szeged, 2011. június 2-4. pp. 11-13 Buczkó, K., Magyari, E.K., Soróczki-Pintér, É., Braun, M., Hubay, K., Tóth, M., Korponai, J.(2011) Response to cooling: Late-glacial and Holocene changes of diatom assemblages and lake acidity in mountain and lowland lakes in the Carpatian Basin. In: Mindrescu, M., Veres, D. & Onac B.P. (eds) Climate change in the Carpathian-Balkan region during late Pleistocene and Holocene. Suseava, Romania, 9-12 June 2011. pp: 39-40. Soróczki-Pintér É., Buczkó K., Magyari E. (2011) Késő-glaciális és Holocén vízszintváltozások a Retyezátban egy gleccsertó kovaalga összetétele alapján. LIII. Hidrobiológus Napok. Tihany, 2011. október 5-7. Soróczki-Pintér É., Buczkó K., Magyari E.: Preuve fondée sur les algues siliceuses de l’augmentation du niveau d’eau et du refroidissement à court terme autour de 9.2-ka dans les Carpates du Sud, Roumanie. In: Edited by: Frédéric Rimet, Agnès Bouchez, Luc Ector & Bernard Montuelle (eds.) ISSN 978-2-7466-6166-0. 32e Colloque de l’ADLaF, and 7th CeDiatom Meeting, Thonon-les-Bains, France, 16-20 Sept. 2013. pp. 77-80. Soróczki-Pintér É. (2013) Fosszilis Chrysophyta sztomatociszták vizsgálata hegyvidéki tavak (Dél-Kárpátok, Románia) üledékszelvényeiben. Pannon Tudományos Nap. Nagykanizsa, 2013. október 17. Soróczki-Pintér É., Magyari E.K., Buczkó K. 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