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In addition, the specificity of primers designed for the amplification of microsatellite loci in <jats:italic>D. dendriticus<\/jats:italic> was tested on <jats:italic>D. ditremus<\/jats:italic> DNA. Due to high identity within the rRNA gene in these species, no suitable DNA regions could be identified for the design of the species-specific primers. A higher level of interspecific differences was detected in the mitochondrial <jats:italic>cox<\/jats:italic>1 and <jats:italic>cob<\/jats:italic> genes, in which regions containing species-specific mutations were chosen for the design of <jats:italic>D. dendriticus<\/jats:italic>- and <jats:italic>D. ditremus<\/jats:italic>-specific primers. However, their specificity was not confirmed, as the <jats:italic>D. dendriticus<\/jats:italic>-specific primers also annealed to <jats:italic>D. ditremus<\/jats:italic> DNA and <jats:italic>vice versa<\/jats:italic>. Of the 15 primer pairs designed for the amplification of microsatellite loci in <jats:italic>D. dendriticus,<\/jats:italic> 13 primer pairs also annealed to <jats:italic>D. ditremus<\/jats:italic> DNA. Only two primer pairs, which amplify the Dd_8 and Dd_33 loci have been proven to be <jats:italic>D. dendriticus<\/jats:italic>-specific. The effectiveness and high reproducibility of the Dd_8 primers were validated on ~3,500 <jats:italic>D. dendriticus<\/jats:italic> and <jats:italic>D. ditremus<\/jats:italic> plerocercoids from Iceland and Norway. These primers are recommended for future molecular differentiation between both <jats:italic>Dibothriocephalus<\/jats:italic> species.<\/jats:p>","DOI":"10.1051\/parasite\/2025033","type":"journal-article","created":{"date-parts":[[2025,7,4]],"date-time":"2025-07-04T07:51:53Z","timestamp":1751615513000},"page":"41","source":"Crossref","is-referenced-by-count":3,"title":["Usefulness of microsatellite loci for differentiating between <i>Dibothriocephalus dendriticus<\/i> and <i>Dibothriocephalus ditremus<\/i> (Cestoda: 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