Three-dimensional morphology of rigid structures as a tool for taxonomic studies of Dactylogyridae (Monogenea)

ROSSIN, MARÍA A.; DE FRANCESCO, PABLO N.; TIMI, JUAN T.

PARASITOLOGY RESEARCH

SPRINGER

Año: 2017

0932-0113

Dactylogyridae is overwhelmingly the most abundant and diverse taxon among monogeneans in continentalwaters of South America. Their small body size requires considerable sampling effort and training for collecting and identifying the worms from the gills, skin, nasal cavities, and othermicrohabitats. Indeed, diagnostic characteristics as scleritesand male copulatory complex are generally less than100-μm long and are essential for taxonomic description andidentification of species. Here, a combination of simple androutine methods for three-dimensional morphological studieson hard structures is proposed for dactylogirids: SDS treatment for clarification of specimens and enzymatic digestionwith proteinase K for freeing sclerotized structures, followedby laser confocal microscopy. This method is applicable tofresh or fixed specimens and does not require staining or dehydration. Indeed, stable autofluorescence emission is detectable at 500?530 nm for bars, anchors, and male copulatorycomplex when excited by argon laser. Advantages of this protocol over previous methodologies for taking laser confocalimages are discussed. Open access software for image processing was used for three-dimensional reconstruction ofsclerotized structures generating models and full 360° rotationvideos