You are here:
  1. University of Vienna
  2. PHAIDRA
  3. Detail o:1310659
Title (eng)
Molecular species determination of larval Digenea as basis for reliable epidemiological analyses, enhanced biodiversity data and significant medical risk assessments
Parallel title (deu)
Molekulare Artenbestimmung von larvalen digenen Trematoden
Author
Larissa Gaub
Advisor
Julia Walochnik
Assessor
Julia Walochnik
Abstract (deu)
Molekulare Artenbestimmung von larvalen digenen Trematoden in Süßwasserschnecken im Bereich der Ufer des Flußes Leitha, von Götzendorf an der Leitha (Niederösterreich) bis Potzneusiedl (Burgenland). Digene Trematoden (=Digenea) beinhalten Arten von medizinischer und veterinärmedizinischer Bedeutung. Die Digenea verfügen über einen komplizierten Lebenszyklus und parasitieren als ersten obligaten Zwischenwirt Mollusken. In den Mollusken entwickeln sich verschiedene Larvenstadien, wie die Sporozysten, Redien und Zerkarien. Diese Larvenstadien sind oftmals mikroskopisch kaum unterscheidbar, was die Artunterscheidung erschwert. Ziel dieser Arbeit war es, das Vorkommen und die Verbreitung von digenen Trematoden in verschiedenen Süßwasserschnecken in den Leitha-Auen, von Götzendorf an der Leitha (Niederösterreich) bis Potzneusiedl (Burgenland), zu ermitteln. Von März bis November 2012, wurden insgesamt 1.184 Süßwasserschnecken, darunter 781 Galba truncatula, 339 Lymnaea stagnalis, 63 Planorbis planorbis und 1 Planorbarius corneus, gesammelt. Insgesamt 66 L. stagnalis, 3 P. planorbis und 1 P. corneus setzten Zerkarien frei. Zusätzlich wurden 781 G. truncatula mikroskopisch auf das Vorhandensein digener Trematoden untersucht. Insgesamt wurden 123 isolierte Trematodenstadien mittels einer PCR (Haider et al., 2012) molekularbiologisch auf Trematoden überprüft. Danach wurden insgesamt 5 Proben mittels Sequenzierung überprüft. Zur Identifizierung der Trichobilharzia Arten wurden die schistosomatiden Zerkarien mittels einer spezifischen PCR getestet (Korsunenko et al., 2010). Für die Identifikation von T. szidati, T. franki und T. regenti wurden die positiven Amplicons nochmal mittels einer PCR getestet (Dvorak et al., 2002) und danach mittels Sequenzierung überprüft. Weder der große amerikanische Leberegel F. magna, noch der gemeine Leberegel F. hepatica konnten in einer der untersuchten Schnecken detektiert werden. Allerdings wurden in drei L. stagnalis Trichobilharzien (3 Trichobilharzia szidati, 1 Trichobilharzia sp.) (Prävalenz: 0.88%) gefunden. Außerdem wurde in einer P. corneus B. polonica nachgewiesen. Nur durch das kontinuierliche Überprüfen des Vorkommens von parasitären Organismen lassen sich zuverlässige epidemiologische Analysen, ordnungsgemäße ökologische Auswertungen, erweiterte Daten über Biodiversität und signifikante medizinische Beurteilung erstellen.
Abstract (eng)
Molecular species determination of the prevalence of larval Digenea in aquatic snail species in the area of the river Leitha, from Götzendorf an der Leitha (lower Austria) to Potzneusiedl (Burgenland). Digenean trematodes (= Digenea) comprise numerous species of medical and veterinary importance. They have complicated life cycles with molluscs serving as obligate first intermediate hosts, inhabiting sporocysts, rediae and cercariae. Often, those stages are morphologically indistinct and therefore not determinable at the species level. The aim of this master thesis was to determine the occurrence of aquatic snails species and to identify their current infection rate with digenean trematodes in the area of the river Leitha, from ‘Götzendorf an der Leitha’ (Lower Austria) to ‘Potzneusiedl’ (Burgenland). Species in focus of the study have been the invasive American Liver Fluke Fascioloides magna, as well as species of the family Schistosomatidae, parasites of birds, the cercariae of which may cause dermatitis in humans and animals. In total, 1,184 samples (781 G. truncatula, 339 L. stagnalis, 63 P. planorbis, 1 P. corneus) of freshwater snails were collected. Of these, 66 individuals of L. stagnalis, 3 individuals of P. planorbis and 1 individual of P. corneus released cercariae. All of these individuals and 53 out of 781 individuals of G. truncatula were examined under the microscope for the presence of digenetic trematodes. A total of 123 samples from affected snails were tested by a trematode universal PCR (Haider et al., 2012). Out of these five samples were further tested by sequencing. For the detection of T.szidati, T. franki and T. regenti, the schistosomatid cercariae (66 samples) were tested with a trichobilharzian specific PCR (Korsunenko et al., 2010). Furthermore, for species identification of the three most common European trichobilharzian species (T. regenti, T. franki, T. szidati) a PCR was chosen, that amplifies the region including ITS1, 5.8S rRNA and ITS2 sequences with the primer pair ITS5/ITS4. Altogehther, 9 out of 66 tested samples were positive and were further tested with this PCR (Dvorak et al., 2002). To sum it up, 7 out of 9 tested samples were positive in this PCR. Altogether, 7 samples were further tested by sequencing after performing the ITS5/ITS4 PCR. By subsequent sequencing of the PCR products and alignment with reference sequences, a species-specific identification of the digenetic specimens was achieved. In short, 83 trematode infections were detected in 1,184 examined G. truncatula, L. stagnalis, P. planorbis and P. corneus. The large American liver fluke F. magna as well as the common liver fluke F. hepatica was not found in the examined G. truncatula. But in three L. stagnalis Trichobilharzia spp. (prevalence 0.88%) were found. Additionally, in one P. corneus B. polonica was found. Cercarial screenings are important to fill data in the databases. Only reliable databases of parasitic organisms warrant reliable epidemiological analyses, proper ecological evaluations, enhanced biodiversity data and significant medical risk assessments.
Keywords (eng)
DigeneaTrematodesAustriaLymnaea stagnalisPlanorbis planorbisPlanorbarius corneusGalba truncatulaTrichobilharzia frankiTrichobilharzia regentiTrichobilharzia szidatiFasciola hepaticaFascioloides magna
Keywords (deu)
DigeneaTrematodenÖsterreichLymnaea stagnalisPlanorbis planorbisPlanorbarius corneusGalba truncatulaTrichobilharzia frankiTrichobilharzia regentiTrichobilharzia szidatiFasciola hepaticaFascioloides magna
Subject (deu)
Parasitologie
Type (deu)
Masterarbeit
Persistent identifier
https://phaidra.univie.ac.at/o:1310659
DOI
10.25365/thesis.33933
URN
urn:nbn:at:at-ubw:1-30011.55279.668369-8
URI
https://utheses.univie.ac.at/detail/30125
Extent (deu)
111 S. : Ill., graf. Darst.
Number of pages
117
Association (deu)
Zentrum für Molekulare Biologie
Members (1)
Title (eng)
Molecular species determination of larval Digenea as basis for reliable epidemiological analyses, enhanced biodiversity data and significant medical risk assessments
Parallel title (deu)
Molekulare Artenbestimmung von larvalen digenen Trematoden
Author
Larissa Gaub
Abstract (deu)
Molekulare Artenbestimmung von larvalen digenen Trematoden in Süßwasserschnecken im Bereich der Ufer des Flußes Leitha, von Götzendorf an der Leitha (Niederösterreich) bis Potzneusiedl (Burgenland). Digene Trematoden (=Digenea) beinhalten Arten von medizinischer und veterinärmedizinischer Bedeutung. Die Digenea verfügen über einen komplizierten Lebenszyklus und parasitieren als ersten obligaten Zwischenwirt Mollusken. In den Mollusken entwickeln sich verschiedene Larvenstadien, wie die Sporozysten, Redien und Zerkarien. Diese Larvenstadien sind oftmals mikroskopisch kaum unterscheidbar, was die Artunterscheidung erschwert. Ziel dieser Arbeit war es, das Vorkommen und die Verbreitung von digenen Trematoden in verschiedenen Süßwasserschnecken in den Leitha-Auen, von Götzendorf an der Leitha (Niederösterreich) bis Potzneusiedl (Burgenland), zu ermitteln. Von März bis November 2012, wurden insgesamt 1.184 Süßwasserschnecken, darunter 781 Galba truncatula, 339 Lymnaea stagnalis, 63 Planorbis planorbis und 1 Planorbarius corneus, gesammelt. Insgesamt 66 L. stagnalis, 3 P. planorbis und 1 P. corneus setzten Zerkarien frei. Zusätzlich wurden 781 G. truncatula mikroskopisch auf das Vorhandensein digener Trematoden untersucht. Insgesamt wurden 123 isolierte Trematodenstadien mittels einer PCR (Haider et al., 2012) molekularbiologisch auf Trematoden überprüft. Danach wurden insgesamt 5 Proben mittels Sequenzierung überprüft. Zur Identifizierung der Trichobilharzia Arten wurden die schistosomatiden Zerkarien mittels einer spezifischen PCR getestet (Korsunenko et al., 2010). Für die Identifikation von T. szidati, T. franki und T. regenti wurden die positiven Amplicons nochmal mittels einer PCR getestet (Dvorak et al., 2002) und danach mittels Sequenzierung überprüft. Weder der große amerikanische Leberegel F. magna, noch der gemeine Leberegel F. hepatica konnten in einer der untersuchten Schnecken detektiert werden. Allerdings wurden in drei L. stagnalis Trichobilharzien (3 Trichobilharzia szidati, 1 Trichobilharzia sp.) (Prävalenz: 0.88%) gefunden. Außerdem wurde in einer P. corneus B. polonica nachgewiesen. Nur durch das kontinuierliche Überprüfen des Vorkommens von parasitären Organismen lassen sich zuverlässige epidemiologische Analysen, ordnungsgemäße ökologische Auswertungen, erweiterte Daten über Biodiversität und signifikante medizinische Beurteilung erstellen.
Abstract (eng)
Molecular species determination of the prevalence of larval Digenea in aquatic snail species in the area of the river Leitha, from Götzendorf an der Leitha (lower Austria) to Potzneusiedl (Burgenland). Digenean trematodes (= Digenea) comprise numerous species of medical and veterinary importance. They have complicated life cycles with molluscs serving as obligate first intermediate hosts, inhabiting sporocysts, rediae and cercariae. Often, those stages are morphologically indistinct and therefore not determinable at the species level. The aim of this master thesis was to determine the occurrence of aquatic snails species and to identify their current infection rate with digenean trematodes in the area of the river Leitha, from ‘Götzendorf an der Leitha’ (Lower Austria) to ‘Potzneusiedl’ (Burgenland). Species in focus of the study have been the invasive American Liver Fluke Fascioloides magna, as well as species of the family Schistosomatidae, parasites of birds, the cercariae of which may cause dermatitis in humans and animals. In total, 1,184 samples (781 G. truncatula, 339 L. stagnalis, 63 P. planorbis, 1 P. corneus) of freshwater snails were collected. Of these, 66 individuals of L. stagnalis, 3 individuals of P. planorbis and 1 individual of P. corneus released cercariae. All of these individuals and 53 out of 781 individuals of G. truncatula were examined under the microscope for the presence of digenetic trematodes. A total of 123 samples from affected snails were tested by a trematode universal PCR (Haider et al., 2012). Out of these five samples were further tested by sequencing. For the detection of T.szidati, T. franki and T. regenti, the schistosomatid cercariae (66 samples) were tested with a trichobilharzian specific PCR (Korsunenko et al., 2010). Furthermore, for species identification of the three most common European trichobilharzian species (T. regenti, T. franki, T. szidati) a PCR was chosen, that amplifies the region including ITS1, 5.8S rRNA and ITS2 sequences with the primer pair ITS5/ITS4. Altogehther, 9 out of 66 tested samples were positive and were further tested with this PCR (Dvorak et al., 2002). To sum it up, 7 out of 9 tested samples were positive in this PCR. Altogether, 7 samples were further tested by sequencing after performing the ITS5/ITS4 PCR. By subsequent sequencing of the PCR products and alignment with reference sequences, a species-specific identification of the digenetic specimens was achieved. In short, 83 trematode infections were detected in 1,184 examined G. truncatula, L. stagnalis, P. planorbis and P. corneus. The large American liver fluke F. magna as well as the common liver fluke F. hepatica was not found in the examined G. truncatula. But in three L. stagnalis Trichobilharzia spp. (prevalence 0.88%) were found. Additionally, in one P. corneus B. polonica was found. Cercarial screenings are important to fill data in the databases. Only reliable databases of parasitic organisms warrant reliable epidemiological analyses, proper ecological evaluations, enhanced biodiversity data and significant medical risk assessments.
Keywords (eng)
DigeneaTrematodesAustriaLymnaea stagnalisPlanorbis planorbisPlanorbarius corneusGalba truncatulaTrichobilharzia frankiTrichobilharzia regentiTrichobilharzia szidatiFasciola hepaticaFascioloides magna
Keywords (deu)
DigeneaTrematodenÖsterreichLymnaea stagnalisPlanorbis planorbisPlanorbarius corneusGalba truncatulaTrichobilharzia frankiTrichobilharzia regentiTrichobilharzia szidatiFasciola hepaticaFascioloides magna
Subject (deu)
Parasitologie
Type (deu)
Masterarbeit
Persistent identifier
https://phaidra.univie.ac.at/o:1310660
Number of pages
117
Association (deu)
Zentrum für Molekulare Biologie
License
All rights reserved
Download