<abstract xml:lang="en" xmlns="http://www.w3.org/1999/xhtml"><p>Molecular comparative analysis of eggs of four liver and stomach flukes of cervids and domestic ruminants, Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, was performed using a new methodological approach for eggshell disintegration. Eggs of all species were crushed mechanically by the Teflon method (PTFE) without use of chemical reagents and an efficient disruption of eggshell was checked microscopically. The egg suspension was then subjected to DNA isolation and PCR amplification using species-specific primers that annealed to the internal transcribed spacer 2 (ITS2) region of ribosomal DNA. The size of PCR products of individual species corresponded well to the size of amplicons obtained from adult flukes. The results provided evidence that the Teflon method does not destroy the structure of egg DNA, thus making the procedure broadly applicable during coprological examinations. Molecular markers introduced here are particularly important for blanket screening and differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs.</p></abstract>

Molecular comparative analysis of eggs of four liver and stomach flukes of cervids and domestic ruminants, Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, was performed using a new methodological approach for eggshell disintegration. Eggs of all species were crushed mechanically by the Teflon method (PTFE) without use of chemical reagents and an efficient disruption of eggshell was checked microscopically. The egg suspension was then subjected to DNA isolation and PCR amplification using species-specific primers that annealed to the internal transcribed spacer 2 (ITS2) region of ribosomal DNA. The size of PCR products of individual species corresponded well to the size of amplicons obtained from adult flukes. The results provided evidence that the Teflon method does not destroy the structure of egg DNA, thus making the procedure broadly applicable during coprological examinations. Molecular markers introduced here are particularly important for blanket screening and differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs.

Molecular discrimination of eggs of cervid trematodes using the Teflon (PTFE) technique for eggshell disruption

Department of Nutrition, Dietetics and Animal Breeding, University of Veterinary Medicine and Pharmacy, Košice, Slovakia

Parasitological Institute, Slovak Academy of Sciences, Košice, Slovakia

Department of Epizootology and Parasitology, University of Veterinary Medicine and Pharmacy, Košice, Slovakia

Helminthologia

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Molecular comparative analysis of eggs of four liver and stomach flukes of cervids and domestic ruminants, Fasciola hepatica, Fascioloides magna, Dicrocoelium...

Molecular discrimination of eggs of cervid trematodes using the Teflon (PTFE) technique for eggshell disruption

Published Online: Oct 09, 2010

Page range: 147 - 151

DOI: https://doi.org/10.2478/s11687-010-0022-y

Keywords
liver and stomach flukes, ruminants, eggs, ribosomal internal transcribed spacer 2, PTFE homogenizer

© 2010 Institute of Parasitology, SAS, Košice, Slovakia

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Molecular discrimination of eggs of cervid trematodes using the Teflon (PTFE) technique for eggshell disruption

Published Online: Oct 09, 2010

Page range: 147 - 151

DOI: https://doi.org/10.2478/s11687-010-0022-y

Keywordsliver and stomach flukes, ruminants, eggs, ribosomal internal transcribed spacer 2, PTFE homogenizer

© 2010 Institute of Parasitology, SAS, Košice, Slovakia

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
liver and stomach flukes, ruminants, eggs, ribosomal internal transcribed spacer 2, PTFE homogenizer