Stimulation of shrimp (Penaeus monodon) hemocytes by lipopolysaccharide-like molecules derived from Novacq™

Abstract

Immune stimulation through feed additives is a promising strategy that can help to combat disease in shrimp farming and reduce the use of antibiotics and other chemotherapeutics. The present study investigated the in vitro immunostimulatory effects of lipopolysaccharide (LPS)-like molecules isolated from the microbial based feed additive Novacq™ (N-LPS). The presence of LPS-like molecules was confirmed and quantified Novacq™ using a HEK-TLR4 reporter cell line. Primary hemocytes isolated from adult Penaeus monodon were used to measure the immunostimulatory of N-LPS compared with the control group that were treated with E. coli derived LPS (E-LPS). The N-LPS stimulated a rapid and significant induction of the phenoloxidase (PO) response in the hemocytes. The PO response increased with exposure time and LPS concentration and was significantly higher compared with an E. coli LPS (E-LPS) control. In addition, using gene expression data, we quantified the transcriptome response of the hemocytes at 15, 30 and 60 mins post stimulation. Compared with the controls, the N-LPS treated hemocytes had a significant up-regulation of genes involved in the immune system modulation and control at all time-points. Most noteworthy was the significant induction of transcripts that function as serine protease inhibitors (namely SERPINs), that regulate the overexpression of the PO system. Transcription factors from the Notch family which directly regulate the expression of many immune genes were also induced within the hemocytes. Additionally, we also saw a strong up-regulation of crustacean hyperglycemic hormone (CHH) transcripts, an important neuropeptide involved in immune function. Overall, the transcriptome profile of the hemocytes suggests that the LPS component of Novacq™ is highly immunostimulatory and generates a strong PO response in vitro. The subsequent transcriptional response appears to be directed towards preventing further activation of the PO system most likely in an attempt to limit cytoxicity to the host. Our study highlights the immunostimulatory ability of Novacq™ and provides further evidence of the positive health benefits this microbial based feed additive can have in shrimp.