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Parazitičtí Helminti Bezocasých Obojživelníků Na Vybraných Lokalitách Iráckého Kurdistánu

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Parazitičtí Helminti Bezocasých Obojživelníků Na Vybraných Lokalitách Iráckého Kurdistánu STŘEDOŠKOLSKÁ ODBORNÁ ČINNOST Obor č. 8: Ochrana a tvorba životního prostředí Parazitičtí helminti bezocasých obojživelníků na vybraných lokalitách Iráckého Kurdistánu Kamila Klementová Jihomoravský kraj Brno 2020 STŘEDOŠKOLSKÁ ODBORNÁ ČINNOST Obor č. 8: Ochrana a tvorba životního prostředí Parazitičtí helminti bezocasých obojživelníků na vybraných lokalitách Iráckého Kurdistánu Parasitic helminths of frogs from selected localities in Iraqi Kurdistan Autoři: Kamila Klementová Škola: Gymnázium Brno, Křenová, příspěvková organizace, Křenová 304/3, 602 00 Brno Kraj: Jihomoravský kraj Konzultant: Mgr. Michal Benovics Brno 2020 Prohlášení Prohlašuji, že jsem svou práci SOČ vypracoval/a samostatně a použil/a jsem pouze prameny a literaturu uvedené v seznamu bibliografických záznamů. Prohlašuji, že tištěná verze a elektronická verze soutěžní práce SOČ jsou shodné. Nemám závažný důvod proti zpřístupňování této práce v souladu se zákonem č. 121/2000 Sb., o právu autorském, o právech souvisejících s právem autorským a o změně některých zákonů (autorský zákon) ve znění pozdějších předpisů. V Brně dne ……………………………………………… Kamila Klementová Poděkování Na tomto místě bych ráda poděkovala rodině i přátelům, kteří mně při psaní této práce podporovali a byli pro mě neskutečnou oporou. Dále děkuji za fotografie odchytových lokalit a hostitelů Mgr. Danielu Jablonskému Ph. D. Největší díky patří vedoucímu mé práce Mgr. Michalu Benovicsovi za poskytnutý materiál, čas, cenné rady, vedení a za připomínky při zpracování této práce. ANOTACE Ve své práci SOČ jsem se zabývala parazitickými organismy žab v oblasti Iráckého Kurdistánu. Cílem bylo identifikovat a zařadit do taxonomických skupin parazity získané z již zmíněných žab. Práce je rozdělena na několik částí. V teoretické části je stručně popsán a rozdělen parazitismus a jsou charakterizovány jednotlivé skupiny parazitických helmintů. Teoretická část se také zaměřuje na obojživelníky obecně a na obojživelníky obývající Irácký Kurdistán. Všechny řády a druhy zmíněných obojživelníků jsou podobně jako helminti charakterizovány. Následuje část praktická, kde je uveden postup fixace a barvení získaných parazitů. Dále je zde popsán postup práce v molekulární laboratoři za účelem získání sekvence DNA pro studium evoluční historie cizopasníků a hostitelů. Hlavním výsledkem mé práce je seznam hostitelů z čeledí Bufonidae a Ranidae s jejich endoparazity. KLÍČOVÁ SLOVA parazitismus; helminti; obojživelníci; hostitelé; parazité ANNOTATION In my SPA I focussed on the parasites of frogs in the Iraqi Kurdistan. The main aim of the SPA was to identify the parasitic helminths collected from the abovementioned frogs. The proposed SPA is divided into two main section: theoretical and practical. The theoretical part encompasses basic description of parasitism, its diversification into multiple classes, and short introduction to higher parasitic taxa. This part focusses also on the amphibians in general (i.e. biology, ecology, morphology and taxonoy) and includes list of amphibians of Iraqi Kurdistan with the brief description of each species present in this region. In the practical part of SPA is described fixation and staining process of collected parasites. Moreover, laboratory procedures for obtaining DNA sequences are also described in this part. The most important output of my SPA is the list of processed host species (representatives of families Bufonidae and Ranidae) and their respective endoparasites. .KEYWORDS parasitism; helminths; amphibians; hosts; parasites Obsah 1 Teoretická část ................................................................................................................................ 8 1.1 Parazitismus ............................................................................................................................. 8 1.2 Parazitičtí helminti ................................................................................................................ 10 1.2.1 Platyhelminthes (ploštěnci) ......................................................................................................... 10 1.2.1.1 Trematoda (motolice) ........................................................................................................................ 12 1.2.1.2 Monogenea (žábrohlísti) .................................................................................................................... 14 1.2.1.3 Cestoda (tasemnice) ........................................................................................................................... 16 1.2.2 Acanthocephala (vrtejši) ............................................................................................................. 17 1.2.3 Nematoda (hlístice) ..................................................................................................................... 19 1.3 Obojživelníci (Amphibia) ...................................................................................................... 21 1.3.1 Gymnophiona (červoři) ............................................................................................................... 22 1.3.2 Caudata (ocasatí) ......................................................................................................................... 23 1.3.3 Anura (žáby) ............................................................................................................................... 25 1.4 Obojživelníci Iráckého Kurdistánu ........................................................................................ 27 1.4.1 Mlokovití (Salamandridae) ......................................................................................................... 27 1.4.1.1 Čolci (Neurergus Cope, 1862) ........................................................................................................... 27 1.4.1.2 Mloci (Salamandra Laurenti, 1768) ................................................................................................... 28 1.4.2 Rosničkovití (Hylidae) ................................................................................................................ 29 1.4.2.1 Rosničky (Hyla Laurenti, 1768) ........................................................................................................ 29 1.4.3 Ropuchovití (Bufonidae) ............................................................................................................ 29 1.4.3.1 Ropuchy (Bufotes Rafinesque, 1815) ................................................................................................ 30 1.4.4 Skokanovití (Ranidae) ................................................................................................................ 30 1.4.4.1 Skokani vodní (Pelophylax Fitzinger, 1843)...................................................................................... 31 2 Materiál ......................................................................................................................................... 32 2.1 Areál zájmu a seznam lokalit ................................................................................................ 32 2.2 Studované druhy .................................................................................................................... 33 3 Metodika ....................................................................................................................................... 34 3.1 Fixace a identifikace hlístic (nematoda) ................................................................................ 34 3.2 Barvení, fixace a identifikace žábrohlístů (monogenea) ....................................................... 34 3.3 Laboratorní postupy .............................................................................................................. 34 3.3.1 Izolace DNA ............................................................................................................................... 34 3.3.2 PCR (polymerázová řetězová reakce) a molekulární analýzy ..................................................... 35 4 Výsledky ....................................................................................................................................... 36 5 Závěr ............................................................................................................................................. 40 6 Použitá literatura……………………………………………………………………………………………………42 7 Seznam obrázků a tabulek …………………………………………………………………………......................45 8 Příloha 1: Fotografie lokalit………………………………………………………………………………………46 9 Příloha 2: Fotografie některých odchycených jedinců……………………………………………………...48 10 Příloha 3: Morfologické znaky hlístic.…………………………………………………………………………50 Úvod Irácký Kurdistán je autonomní oblast Kurdů o rozloze asi 36 000 km² v severovýchodním Iráku. Má od roku 1991 vlastní vládu, parlament i armádu, ale není plně nezávislá [1]. Lokální fauna je relativně chudá na počet druhů obojživelníků. V průběhu posledních let byl zaznamenaný narůstající úbytek žab po celém světě, což zvyšuje pozornost na studium ekologie obojživelníků [2]. Jeden z důležitých aspektů působících na ekologii žab jsou parazitické organismy. Paraziti ovlivňují organizmus hostitele různými způsoby a to hlavně na jeho chování, rozmnožovaní a zhoršují zdravotní stav [3]. Výstup z mé práce by měl rozšířit poznatky o lokální herpetologii (nauce o studenokrevných čtvernožcích) a jejich cizopasnících, jež jsou potřebné pro účely ochrany biodiverzity. Cíle práce: 1. Vytvořit stručnou charakteristiku jednotlivých
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