Introduction
Shrimp are one of the most important groups of animals in global aquaculture. The Pacific whiteleg shrimp (Litopenaeus vannamei ) is the most cultured shrimp species, accounting for over 50% of worldwide production. Unfortunately, the industry’s efficiency and sustainability are threatened by significant annual losses due to infectious diseases, 60% of which are caused by viral pathogens such as white spot syndrome virus (WSSV). Efforts to limit the impact of disease caused by this pathogen are impeded by a lack of effective treatments available.
Materials and methods
In order to establish whiteleg shrimp primary cell cultures, we extracted haemolymph, lymphoid organ and hepatopancreas samples from sterile shrimp and cultured them as dissociated cells or explants in a range of cell media mixes while monitoring the growth and survivability of cells. For RNA sequencing, we isolated nuclei from snap-frozen hepatopancreas tissue and processed it using 10x Chromium Next GEM Single Cell Dual Index kits. The sequenced samples have been analysed and the cell atlas was built using cell cluster marker genes.
Results
Our team has established an in vivo infection model in juvenile and adult stage shrimp and developed primary cell culture systems for the study of host response to WSSV using haemocytes, hepatopancreas and lymphoid organ tissues from adult L. vannamei (Figure 1) . Additionally, we have establish a novel protocol for nuclei isolation and single nuclei RNA-sequencing, and in doing so, create a hepatopancreas cell atlas for the species (Figure 2) .
Conclusion
The novel protocols will help us use high throughput transcriptomic analysis to identify priority candidate genes for WSSV resistance via single-nuclei RNA sequencing of WSSV-infected and non-infected shrimp. The results will serve as a foundation for future studies using in vitro models and single nuclei sequencing in whiteleg shrimp and set a new foundation for developing therapeutic strategies to combat WSSV in shrimp aquaculture.