Transcriptome analyses of mRNA and circular RNA reveal dietary supplementation with freeze-dried Lactiplantibacillus plantarum primes immune memory of Whiteleg shrimp (Penaeus vannamei) against pathogens

The lack of a classical adaptive immunity renders the development of disease control and prevention measures in shrimp challenging. In this study, the concept of trained immunity was exploited in the development of a feed supplement. Penaeus vannamei shrimp was fed with feed supplemented with freeze-dried whole culture of Lactiplantibacillus plantarum (FD-LAB) for 15 days. RNA sequencing using Illumina platform was performed on the gill and stomach tissues collected at specific time points during the feeding period (0^th day, 8th day, 15th day). Differentially-expressed genes (DEGs) previously reported to have innate immunity- and immune memory-related functions were selected for validation. Additionally, the differential expression of putatively immune-related circular RNAs (DECs) were also explored as these noncoding regulatory RNAs may also influence host immunity. Challenge tests with either the acute hepatopancreatic necrosis disease-causing strain Vibrio parahaemolyticus D6 or White Spot Syndrome Virus (WSSV) were conducted. Transcriptome analyses showed that FD-LAB supplementation resulted to DEGs and DECs related to pathogen recognition, antimicrobial peptides, transcription regulation, and immune memory. Challenge tests performed immediately after 15 days and 8 days of feeding showed protection on P. vannamei by FD-LAB against bacterial and viral pathogens. Increase in survival rates were also observed upon challenge with both pathogens 7 days and 14 days after last intake of FD-LAB, indicating trained immunity in shrimp. Our study highlighted the effects of FD-LAB on the innate immunity and immune memory of P. vannamei against bacterial and viral pathogens. These findings emphasize the possibility of immunostimulants inducing lasting enhanced immunity against infections despite the lack of a classical adaptive immunity in shrimp.