Szirányi Barbara Mária

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Szirányi Barbara Mária EÖTVÖS LORÁND TUDOMÁNYEGYETEM TERMÉSZETTUDOMÁNYI KAR BIOLÓGIA DOKTORI ISKOLA ZOOTAXONÓMIA, ÁLLATÖKOLÓGIA, HIDROBIOLÓGIA DOKTORI PROGRAM SZIRÁNYI BARBARA MÁRIA ENERGETIKAI CÉLOKRA HASZNÁLT TERMÁLVIZEK BEFOGADÁSÁRA SZOLGÁLÓ TÁROZÓTAVAK MIKROBIÁLIS DIVERZITÁSA -doktori értekezés- Témavezető Dr. Borsodi Andrea habil. egyetemi docens Biológiai Doktori Iskola vezetője Prof. Dr. Erdei Anna Zootaxonómia, állatökológia, hidrobiológia doktori program vezetője Prof. Dr. Török János ELTE TTK Biológiai Intézet, Mikrobiológiai Tanszék BUDAPEST 2019 Tartalomjegyzék 1. Bevezetés ............................................................................................................................................. 1 2. Irodalmi áttekintés ............................................................................................................................... 2 2.1 Magyarország geotermális adottságai és azok kihasználtsága ....................................................... 2 2.2 Használat utáni elhelyezés, visszasajtolási lehetőségek és korlátozó tényezők ............................. 5 2.3 A szarvasi Barex Kft. által üzemeltetett termelőkutak vizének kémiai és mikrobiológiai tulajdonságai ........................................................................................................................................ 8 2.4 A szarvasi Therm-Organ-tó mikrobiológiai vizsgálatának eredményei ........................................... 9 2.5 Alkalikus-szalinikus sekély tavak mikrobiológiája ......................................................................... 11 2.6 Fenolvegyületek biodegradációja alkalikus-szalinikus környezetekben ....................................... 19 3. Célkitűzések ....................................................................................................................................... 23 4. Anyag és módszer .............................................................................................................................. 24 4.1 A vizsgálati helyszín bemutatása .................................................................................................. 24 4.2 Mintavételezés............................................................................................................................. 25 4.3 A vizsgált évekre jellemző időjárási viszonyok ............................................................................. 28 4.4 Vízminőségi vizsgálatok ................................................................................................................ 29 4.5 Epifluoreszcens mikroszkópos sejtszám meghatározás ............................................................... 30 4.6. Tenyésztésen alapuló vizsgálatok ............................................................................................... 30 4.6.1 Baktériumtörzsek izolálása.................................................................................................... 30 4.6.2 A baktériumtörzsek genotípusos vizsgálata .......................................................................... 30 4.6.3 ARDRA alapú csoportosítás ................................................................................................... 31 4.6.4 A DNS bázissorrendjének meghatározása és a szekvenciák értékelése ................................ 32 4.7 Tenyésztéstől független közösségszerkezeti vizsgálatok ............................................................. 33 4.7.1 Mintaelőkészítés és közösségi DNS izolálás .......................................................................... 33 4.7.2 Denaturáló gradiens gélelektroforézis (DGGE) ..................................................................... 33 4.7.3 Molekuláris klóntárak létrehozása és feldolgozása ............................................................... 35 4.7.4 A mikrobaközösségek összetételének feltárása piroszekvenálással ..................................... 37 4.7.5 Adataink statisztikai elemzése és az alkalmazott diverzitás indexek .................................... 39 5. Eredmények ismertetése ................................................................................................................... 41 5.1 A vízminőségi vizsgálatok eredményei ......................................................................................... 41 5.2 Az epifluoreszcens mikroszkópián alapuló sejtszámlálás eredményei ......................................... 46 5.3 A tenyésztésen alapuló vizsgálatok eredményei .......................................................................... 47 5.3.1 Csíraszámbecslés .................................................................................................................. 47 5.3.2 A baktériumtörzsek azonosítása ........................................................................................... 47 5.4 A denaturáló gradiens gélelektroforézis (DGGE) vizsgálatok eredményei ................................... 50 5.5 A molekuláris klóntárak feldolgozásának eredményei ................................................................. 55 5.5.1 A molekuláris klóntárak alfa-diverzitása ............................................................................... 56 5.5.2 A molekuláris klóntárak béta-diverzitása .............................................................................. 58 5.5.3 A molekuláris klóntárak taxonómiai diverzitása .................................................................... 60 5.6 A piroszekvenálással végzett közösségfeltárás eredményei ........................................................ 68 5.6.1 Az amplikontárak alfa-diverzitása ......................................................................................... 68 5.6.2 Az amplikontárak béta-diverzitása ........................................................................................ 69 5.6.3 Az amplikontárak filogenetikai diverzitása ............................................................................ 71 6. Eredmények értékelése ..................................................................................................................... 81 6.1 A tórendszerben bekövetkező tér- és időbeli változások, és a tavak közötti közösségszerkezeti eltérések ............................................................................................................................................ 81 6.2 A taxonómiai, különös tekintettel az extremofil (alkalofil, halofil) szervezetek diverzitásának összevetése hasonló élőhelyekével ................................................................................................... 87 6.3 Anyagátalakítási folyamatok a tározótó rendszerben .................................................................. 95 6.3.1 Szén körforgalom .................................................................................................................. 95 6.3.2 Nitrogén körforgalom ......................................................................................................... 100 6.3.3 Kén körforgalom ................................................................................................................. 102 7. Összegzés ......................................................................................................................................... 106 7.1 Következtetések és továbblépési lehetőségek ........................................................................... 110 8. Kivonat ............................................................................................................................................. 112 9. Abstract ............................................................................................................................................ 113 10. Köszönetnyilvánítás ....................................................................................................................... 114 11. Idézett irodalom ............................................................................................................................ 115 12. Függelék ......................................................................................................................................... 124 DOI adatlap .......................................................................................................................................... 131 1. Bevezetés Hazánk termálvízadottságait tekintve Európában vezető helyen áll; jelentős mennyiségű alacsony és közepes entalpiájú, közvetlen hasznosításra és villamosenergia-termelésre alkalmas geotermális potenciállal rendelkezik (Árpási 1992, 1998, 2003; Árpási és Szabó 1999; Bobok, Mating, és Navratil 1988; Boldizsár 1967; Korim 1998). Magyarországon a közvetlen használatra termelt termálvizek mennyisége 2002-ben hozzávetőleg 22 millió m3 volt, 31 °C-os átlagos felhasználási hőmérséklettel. A geotermális energia fő fogyasztója a mezőgazdaság (Árpási 2003). A hazai termálvizek túlnyomó része alkáli-hidrogénkarbonátos, alkáli-karbonátos víz. Kémiai és biológiai oxigénigényük a redukált állapotú szerves anyagok és szervetlen vegyületek magas koncentrációja miatt jelentős (Pekár 2008). A Kárpáti és mtsai (1999) által vizsgált termálvizek mindegyike többféle poliaromás szénhidrogént, sok esetben fenolokat tartalmazott. A használt termálvíz a természetes vizekhez képest magasabb hőmérséklete, pH-ja, só- és fenoltartalma miatt nem vezethető vissza közvetlenül a felszíni élővizekbe (Pekár 2008), így a felhasznált vízmennyiség döntő hányadát ún. hűtőtavas átmeneti tározás után vezetik felszíni folyóvízbe, általában
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