A staining method for detection of Enterocytozoon hepatopenaei (EHP) spores with calcofluor white
Ruo-Heng Zhao1, Wen Gao2, Liang Qiu1
1Laboratory 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.
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.
Insights
Calcofluor white (CFW) staining offers a simple and rapid method for detecting Enterocytozoon hepatopenaei (EHP) spores. This fluorescent dye effectively visualizes EHP spores in shrimp samples without staining host tissues.
Area of Science:
- Aquatic animal health
- Microbiology
- Parasitology
Background:
- Enterocytozoon hepatopenaei (EHP) is a microsporidian parasite affecting shrimp.
- Accurate detection of EHP is crucial for managing shrimp aquaculture.
Purpose of the Study:
- To evaluate calcofluor white (CFW) as a diagnostic stain for EHP spores.
- To establish a rapid and reliable method for EHP detection in Penaeus vannamei.
Main Methods:
- Utilized calcofluor white (CFW), a chitin and cellulose binding fluorescent dye.
- Applied CFW staining to EHP spores in suspension, feces, and infected shrimp hepatopancreas.
- Compared CFW staining with hematoxylin and phloxine (H&P) staining for EHP detection.
Main Results:
- CFW staining clearly visualized EHP spores with distinct blue-white fluorescent oval walls.
- CFW did not stain the host shrimp tissues, ensuring specific detection of EHP.
- Hematoxylin and phloxine (H&P) staining showed EHP spores as orange-red spots in tissue sections.
- CFW staining proved to be a simple and rapid method for EHP identification.
Conclusions:
- Calcofluor white (CFW) is an effective and specific stain for detecting Enterocytozoon hepatopenaei (EHP) spores.
- CFW staining provides a valuable tool for the rapid diagnosis of EHP in shrimp aquaculture.
- This method simplifies EHP detection in various sample types, including feces and infected tissues.
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