Identification of Metastrongylidae species occurring in wild boar
AbstractLungworms (Nematoda, Metastrongylidae) occurring in the Suidae may be divided in two groups according to their phenotypic traits. The first group includes species whose females have provaginas in the posterior part of the body (M. pudendotectus, M. asymmetricus), whilst in the second group of species, females do not have provaginas (M. elongatus, M. confusus, M. salmi). The size of the provagina varies within a single species and is often characterized by a wide inter-individual variability. Similarly, there may be difficulties in identifying females from the second group. Furthermore, the dimensions of the parasites are subject to a high degree of divergence, and some species ranges overlap. Observing the terminations of copulatory spicules and the shape of the male copulatory bursa is the most authoritative means of phenotypic classification of parasites of the Metastrongylus genus. However, the recognition of male specimens does not conclusively identify the species of parasites present in one hosts, because their invasion has mixed nature. Hence, the molecular identification of parasites of Metastrongylus genus occurring in wild boars in Lower Silesia was attempted to eliminate potential errors resulting from the imperfections of the human eye. Seven females of lungworm species from Metastrongylidae family isolated from wild boar lungs were subjected to molecular examination. The wild boar was shot in the Zlotowek Forest Inspectorate in March 2012. The genetic material of parasites was isolated using a commercial test for DNA isolation from tissue (A&A Biotechnology). A flanking sequence was amplified within the ITS-2 region, and the obtained PCR products were divided into two parts. The first part of the DNA matrix was sequenced and the results of the analysis were compared with the GenBank library. The second part of the genetic material was analyzed with the use of the RAPD method. It is a random amplification of polymorphic DNA, which relies on the random amplification of the short DNA fragments. The short primers for the study were selected based on the available literature. The DNA of different parasites species within the Metastrongylus genus is varied. Striped patterns characteristic of each individual species were received after electrophoresis of the PCR product on agarose gel. The affinity of the examined parasites can be assessed thanks to the similarity of the striped patterns obtained in this test. The study was conducted in two ways so that a comparison of the results will allow the costly sequencing process of genetic material of each isolated individual to be bypassed in the future.
|Journal series||Annals of Parasitology, ISSN 2299-0631, e-ISSN 2300-6706, (B 15 pkt)|
|Conference||XXIIIth Congress of the Polish Parasitological Society, 04-09-2013 - 07-09-2013, Szklarska Poręba-Piechowice, Polska|
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