A Simple and Rapid Staining Technique for Sex Determination of Trichinella Larvae Parasites by Confocal Laser Scanning Microscopy

• Published in: Microscopy and microanalysis Vol. 25; no. 6; pp. 1491 - 1497
• Main Authors: Gavarane, Inese, Kirilova, Elena, Rubeniņa, Ilze, Mežaraupe, Ligita, Osipovs, Sergejs, Deksne, Gunita, Pučkins, Aleksandrs, Kokina, Inese, Bulanovs, Andrejs, Kirjušina, Muza
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01.12.2019
• Subjects: Chromatography; Confocal microscopy; Dyes; Ethanol; Females; Fixatives; Hydrocarbons; Inspection; Intestine; Larvae; Life cycles; Males; Meat; Microscopy; Morphology; Muscles; Nematodes; Nervous system; NMR; Nuclear magnetic resonance; Parasites; Reproductive system; Sample preparation; Scanning microscopy; Sex; Sex determination; Spectrum analysis; Staining; Studies; Synthesis; Trichinella; Trichinellosis; Writing; Zoonoses; Japan; benzanthrone dye; Bouin's fixative; Trichinella larvae sex determination; confocal laser scanning microscopy; fixatives
• ISSN: 1431-9276; 1435-8115
• DOI: 10.1017/S1431927619015046

The roundworms of Trichinella genus are worldwide distributed and their prevalence in nature is high. Trichinella genus parasites are the causative agents of foodborne zoonosis trichinellosis. The main prevention and control of the infection are meat inspection by the magnetic stirrer method for the detection of Trichinella larvae in muscle samples. The treatment can be effective if the parasite is discovered early in the intestinal phase. Once the Trichinella larva has reached the muscle tissue, the parasite remains therein and there is no treatment for this life cycle stage. The Trichinella species is dioecious with separate male and female individuals. The developed staining technique that uses confocal laser scanning microscopy (CLSM) displays sufficient results for Trichinella larvae examination and this protocol is applicable to study the internal and external structures and for the sex determination of T. britovi and T. spiralis larvae samples. In the present study, a luminescent derivative was synthesized and used for staining of T. spiralis and T. britovi larvae samples for the examination by CLSM. Various fixatives, such as AFA, 70% ethanol, and Bouin's and Carnoy's solutions were tested for sample preparation. The synthesized luminescent compound demonstrates best visualization results for samples fixed in Bouin's fixative.