An improved method for isolating camallanid (Nematoda) spicules for scanning electron microscopy.

Abstract

In nematodes, the structure of male copulatory organs is a significant taxonomic distinguisher and includes the morphometry of the spicules. The description of these structures mainly relies on the study of whole mounts using light microscopy. In rare instances, protruding spicules have been described with scanning electron microscopy. Even fewer studies have described the ultrastructure of isolated spicules following their isolation. In the present study, two different methods of spicule isolation were performed on two parasitic camallanid nematodes, Procamallanus (Procamallanus) pseudolaeviconchus Moravec & van As, 2015 and Paracamallanus cyathopharynx (Baylis, 1923), from African sharptooth catfish to determine the practicality and efficiency of the methodologies. The first method involved using sharpened tungsten needles and microdissection of the spicule pouch to free the spicules, followed by soft tissue digestion if necessary. Alternatively, the spicules were isolated through mechanical release instead of dissection in a method developed in the current study. This involved freeing the spicules from surrounding soft tissue by placing live specimens between a coverslip and a glass slide in a drop of water and exerting pressure with small rotational movements. Both methods yielded favourable results, but Method 2 is recommended for future studies due to the many advantages.