engleski

Determination of Acanthocephala parasites in European chub (Leuciscus cephalus) from Sava River based on mitochondrial DNA, 18S and ITS rDNA sequence analysis

Acanthocephalans are wide distributed, spiny-headed endoparasitic worms with complex life cycle including broad range of intermediate and definitive (e.g. fish) hosts. Due to difficulty in determination of morphologically similar strains, several DNA methods were developed for studies on the systematics of acanthocephalans. During the two year period of health inspection of European chub (Leuciscus cephalus) in Sava River, significant number of examined fish appeared to be infected with acanthocephalans. Subsequently, the study to determine the species level of detected parasites was undertaken. Acanthocephalans were sampled from the intestine of European chub caught at four locations of Sava River. Total DNA extraction from individuals was done and PCR was used to amplify partial nucleotide sequences of: cytochrome oxidase subunit I (COI), ribosomal gene 18S and ribosomal internal transcribed spacers (ITS) regions. Due to possible problems with nuclear mitochondrial pseudogens, RNA extraction and subsequent RT-PCR was applied as control for COI gene amplification. Determined sequences of PCR products were compared to the similar reported in GenBank. The percentage of infected fish at different sampling locations in Sava River ranged from 36, 0% to 77, 6%. Preliminary results confirmed the presence of Pomphorhynchus laevis and Acanthocephalus dirus in analyzed fish. High prevalence of P. laevis in European chub demonstrated in this study suggests possible use of this parasite as bioindicator for metal pollution in Sava River.

Acanthocephala; European chub; cytochrome oxidase subunit I; 18S rDNA; ITS regions

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