An improved method for isolating camallanid (Nematoda) spicules for scanning electron microscopy. 2023

A P Nofal, and Q M Dos Santos, and A Avenant-Oldewage
Department of Zoology, University of Johannesburg, PO Box 524, Auckland Park, Johannesburg 2006, South Africa.

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.

UI MeSH Term Description Entries
D008297 Male Males
D008855 Microscopy, Electron, Scanning Microscopy in which the object is examined directly by an electron beam scanning the specimen point-by-point. The image is constructed by detecting the products of specimen interactions that are projected above the plane of the sample, such as backscattered electrons. Although SCANNING TRANSMISSION ELECTRON MICROSCOPY also scans the specimen point by point with the electron beam, the image is constructed by detecting the electrons, or their interaction products that are transmitted through the sample plane, so that is a form of TRANSMISSION ELECTRON MICROSCOPY. Scanning Electron Microscopy,Electron Scanning Microscopy,Electron Microscopies, Scanning,Electron Microscopy, Scanning,Electron Scanning Microscopies,Microscopies, Electron Scanning,Microscopies, Scanning Electron,Microscopy, Electron Scanning,Microscopy, Scanning Electron,Scanning Electron Microscopies,Scanning Microscopies, Electron,Scanning Microscopy, Electron
D009348 Nematoda A phylum of unsegmented helminths with fundamental bilateral symmetry and secondary triradiate symmetry of the oral and esophageal structures. Many species are parasites. Phasmidia,Secernentea,Sipunculida
D002397 Catfishes Common name of the order Siluriformes. This order contains many families and over 2,000 species, including venomous species. Heteropneustes and Plotosus genera have dangerous stings and are aggressive. Most species are passive stingers. Eremophilus mutisii,Heteropneustes,Plotosus,Siluriformes,Arius,Catfish,Colombian Catfish,Catfish, Colombian
D005618 Fresh Water Water containing no significant amounts of salts, such as water from RIVERS and LAKES. Freshwater,Fresh Waters,Freshwaters,Water, Fresh,Waters, Fresh
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D017174 Spirurida An order of nematodes of the subclass SECERNENTEA. Characteristics include ventral or caudoventral papillae and an esophagus divided into anterior muscular and posterior glandular parts. Spiruridas

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