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

Part of: Micrographia Collection

Published online by Cambridge University Press:  28 October 2019

Inese Gavarane Open the ORCID record for Inese Gavarane [Opens in a new window] ,
Elena Kirilova ,
Ilze Rubeniņa ,
Ligita Mežaraupe ,
Sergejs Osipovs ,
Gunita Deksne ,
Aleksandrs Pučkins ,
Inese Kokina ,
Andrejs Bulanovs  and
Muza Kirjušina
Inese Gavarane*
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Elena Kirilova
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Ilze Rubeniņa
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Ligita Mežaraupe
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Sergejs Osipovs
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Gunita Deksne
Affiliation:
Institute of Food Safety, Animal Health and Environment BIOR, Riga LV-1076, Latvia Faculty of Biology, University of Latvia, Riga LV-1004, Latvia
Aleksandrs Pučkins
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Inese Kokina
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Andrejs Bulanovs
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
Muza Kirjušina
Affiliation:
Institute of Life Sciences and Technology, Daugavpils University, Daugavpils LV-5401, Latvia
*
*Author for correspondence: Inese Gavarāne, E-mail: inese.gavarane@du.lv
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Abstract

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.

Keywords

benzanthrone dyeBouin's fixativeconfocal laser scanning microscopyfixativesTrichinella larvae sex determination
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 25 , Issue 6 , December 2019 , pp. 1491 - 1497
Copyright
Copyright © Microscopy Society of America 2019 

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