Detection of the intranuclear microsporidian Enterospora nucleophila in gilthead sea bream by in situ hybridization.
Nahla Hossameldin AhmedMonica CaffaraAriadna Sitjà-BobadillaMaria Letizia FioravantiAngelica MazzoneAbbass Sayed AboulezzAsmaa Mohamed MetwallyMosaab Adl-Eldin OmarOswaldo R PalenzuelaPublished in: Journal of fish diseases (2019)
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.
Keyphrases
- plasmodium falciparum
- toxoplasma gondii
- trypanosoma cruzi
- induced apoptosis
- peripheral blood
- life cycle
- cell cycle arrest
- endoplasmic reticulum stress
- high intensity
- single cell
- signaling pathway
- oxidative stress
- cell death
- cell therapy
- stem cells
- fluorescence imaging
- cell proliferation
- photodynamic therapy
- solid phase extraction
- nucleic acid
- molecularly imprinted
- simultaneous determination
- loop mediated isothermal amplification
- clinical evaluation