Trichinella spp

Nicholas, Bachand, Ravel, A., Leighton, P., Stephen, C., Iqbal, A., Ndao, M., Konecsni, K., Fernando, C. & Jenkins, E., 2018, Foxes (Vulpes vulpes) as sentinels for parasitic zoonoses, Toxoplasma gondii and Trichinella nativa, in the northeastern Canadian Arctic, International Journal for Parasitology: Parasites and Wildlife 7 (3), pp. 391-397 : 394

2.6. Detection and genetic characterization of Trichinella spp

Tongues were collected from each fox and frozen at −20 ̊C for <2 months until Trichinella larvae were recovered using a double centrifugation and enzymatic digestion assay described by Forbes and Gajadhar (1999). Briefly, approximately 10 g from each tongue was homogenized in a blender, digested in 1% HCl/Pepsin solution at 37 ̊C for 1 h, and concentrated by sequential sedimentation through two different separatory funnels. Sediment was examined under a stereomicroscope at 10-16× magnification for the presence of larvae. Recovered larvae were washed in PBS and frozen at minus 20 ̊C until further analysis. For each positive fox, DNA was extracted from 15 larvae (5 individually and 10 pooled) for further characterization as per Scandrett et al. (2018). The extracted DNA was amplified by a conventional multiplex PCR using primer sets that generate unique banding patterns on a 2.5% agarose gel for each known species and genotype of Trichinella ( Zarlenga et al., 1999) .