Rapid identification of nine species of diphyllobothriidean tapeworms by pyrosequencing

Abstract

The identification of diphyllobothriidean tapeworms (Cestoda: Diphyllobothriidea) that infect humans and intermediate/paratenic hosts is extremely difficult due to their morphological similarities, particularly in the case of Diphyllobothrium and Spirometra species. A pyrosequencing method for the molecular identification of pathogenic agents has recently been developed, but as of yet there have been no reports of pyrosequencing approaches that are able to discriminate among diphyllobothriidean species. This study, therefore, set out to establish a pyrosequencing method for differentiating among nine diphyllobothriidean species, Diphyllobothrium dendriticum, Diphyllobothrium ditremum, Diphyllobothrium latum, Diphyllobothrium nihonkaiense, Diphyllobothrium stemmacephalum, Diplogonoporus balaenopterae, Adenocephalus pacificus, Spirometra decipiens and Sparganum proliferum, based on the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene as a molecular marker. A region of 41 nucleotides in the cox1 gene served as a target, and variations in this region were used for identification using PCR plus pyrosequencing. This region contains nucleotide variations at 12 positions, which is enough for the identification of the selected nine species of diphyllobothriidean tapeworms. This method was found to be a reliable tool not only for species identification of diphyllobothriids, but also for epidemiological studies of cestodiasis caused by diphyllobothriidean tapeworms at public health units in endemic areas.

The Intestinal Eukaryotic and Bacterial Biome of Spotted Hyenas: The Impact of Social Status and Age on Diversity and Composition

“…and Diphyllobothrium spp. based on egg morphology is challenging (Thanchomnang et al, 2016 ). Our sequencing based taxonomic assignments suggest that eggs from both Spirometra spp.…”

Intestinal parasites at the Late Bronze Age settlement of Must Farm, in the fens of East Anglia, UK (9th century B.C.E.)

“…Therefore, it seems likely that both D. latum and D. dendriticum were endemic in Late Bronze Age Britain. Genetic analysis of these species is currently the best way to generate an accurate species identification (Wicht et al, 2010;Kuchta et al, 2015;Thanchomnang et al, 2016). Identification of the species of whipworm (Trichuris sp.)…”

Diphyllobothriidae

“…pacificus, Sp. decipiens, and Sp. proliferum has been described based on the mitochondrial cytochrome c oxidase subunit 1(cox1) gene as a molecular marker (Thanchomnang et al, 2016).…”

“…Infection in native fish from Chile, such as Galaxias spp., Percichthys trucha, Odontesthes mauleanum, and Basilichthys australis have only been observed in 2 lakes with a low intensity of infection in contrast to the high parasitic load of introduced rainbow trout (Torres et al, 1989b(Torres et al, , 1998(Torres et al, , 2004a(Torres et al, , 2012. Andersen and Gibson, 1989;De Vos and Dick, 1989;Halvorsen and Wissler, 1973;Janicki and Rosen, 1917;Kuchta et al, 2015a;Mercado et al, 2010;Revenga, 1993;Scholz et al, 2009;Thanchomnang et al, 2016;Torres et al, 1989b, 1998, 2004ab Bylund, 1972Chubb, 1980;De Vos and Dick, 1989;Gustafsson and Vaihela, 1981;Henricson, 1977;Kuchta et al, 2013;Moravec, 2004;Revenga, 1993;Thanchomnang et al, 2016;Torres, 1990;Torres et al, 1981bTorres et al, , 1989bTorres et al, , 1998Torres et al, , 2004aTorres et al, , 2012Vik, 1957 c Ando et al, 2001;Fang et al, 2015;Jeon et al, 2009;Kuchta et al, 2017;Suzuki et al, 2010;Yamane et al,1986;Yanagida et al, 2010d Escalante, 1983Escalante and Miranda, 1986;...…”