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Detection of the intranuclear microsporidian Enterospora nucleophila in gilthead sea bream by in situ hybridization
Author(s) -
Ahmed Nahla Hossameldin,
Caffara Monica,
SitjàBobadilla Ariadna,
Fioravanti Maria Letizia,
Mazzone Angelica,
Aboulezz Abbass Sayed,
Metwally Asmaa Mohamed,
Omar Mosaab AdlEldin,
Palenzuela Oswaldo R.
Publication year - 2019
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.12993
Subject(s) - parasite hosting , biology , spore , microsporidia , in situ hybridization , microsporidiosis , in situ , oligomer restriction , microbiology and biotechnology , oligonucleotide , gene , gene expression , physics , world wide web , computer science , meteorology , biochemistry
Enterospora nucleophila is an intranuclear microsporidian responsible for emaciative microsporidiosis of gilthead sea bream (GSB). Its minute size and cryptic nature make it easily misdiagnosed. An in situ hybridization (ISH) technique based on antisense oligonucleotide probes specific for the parasite was developed and used in clinically infected GSB in combination with calcofluor white stain (CW) and other histopathological techniques. The ISH method was found to label very conspicuously the cells containing parasite stages, with the signal concentrating in merogonial and sporogonial plasmodia within the infected cell nuclei. Comparison with CW demonstrated limited ISH signal in cells containing mature spores, which was attributed mostly to the scarcity of probe targets present in these stages. Although spores were detected in other organs of the digestive system as well as in the peripheral blood, proliferative stages or parasite reservoirs were not found in this work outside the intestines. The study demonstrated a frequent disassociation between the presence of abundant spores and the intensity of the infections as determined by the parasite activity. The ISH allows confirmatory diagnosis of GSB microsporidiosis and estimation of infection intensity and will be a valuable tool for a more precise determination of parasite dissemination pathways and pathogeny mechanisms.