A staining method for detection of Enterocytozoon hepatopenaei (EHP) spores with calcofluor white

Ruo-Heng Zhao; Wen Gao; Liang Qiu; Xing Chen; Xuan Dong; Chen Li; Jie Huang

doi:10.1016/j.jip.2020.107347

A staining method for detection of Enterocytozoon hepatopenaei (EHP) spores with calcofluor white

J Invertebr Pathol. 2020 May:172:107347. doi: 10.1016/j.jip.2020.107347. Epub 2020 Feb 28.

Authors

Ruo-Heng Zhao¹, Wen Gao², Liang Qiu¹, Xing Chen¹, Xuan Dong¹, Chen Li¹, Jie Huang³

Affiliations

¹ Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
² Shanghai Ocean University, Shanghai 201306, China; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
³ Shanghai Ocean University, Shanghai 201306, China; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China. Electronic address: [email protected].

PMID: 32119955
DOI: 10.1016/j.jip.2020.107347

Abstract

A fluorescent stain, calcofluor white (CFW), was used for detection of spores of the microsporidium Enterocytozoon hepatopenaei (EHP). EHP spores in suspension, in feces, or in the infected hepatopancreas of the shrimp Penaeus vannamei, can be easily stained with this chitin and cellulose binding dye to show distinct blue-white fluorescent oval walls. The dye does not stain the host tissues. EHP spores showed orange-red spots by staining with hematoxylin and phloxine (H&P) in the section. CFW staining provides a simple and rapid method for determining the presence of EHP spores in fecal or tissue samples.

Keywords: Calcofluor white stain; Enterocytozoon hepatopenaei; Fluorescence; Histopathology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Benzenesulfonates / chemistry*
Enterocytozoon / isolation & purification*
Feces / microbiology
Hepatopancreas / microbiology
Penaeidae / microbiology*
Spores, Fungal / isolation & purification*
Staining and Labeling / methods*

Substances

Benzenesulfonates
C.I. Fluorescent Brightening Agent 28

Supplementary concepts

Enterocytozoon hepatopenaei