Effects of Vibro harveyi and Staphyloccocus aureus infection on hemocyanin synthesis and innate immune responses in white shrimp Litopenaeus vannamei

Abstract

Hemocyanin, a multifunctional oxygen-carrying protein, has critical effects on immune defense in crustaceans. To explore the role of hemocyanin in anti-pathogen mechanism, effects of Vibrio harveyi (V. harvey) and Staphyloccocus aureus (S. aureus) on hemocyanin synthesis and innate immune responses were investigated in Litopenaeus vannamei (L. vannamei) during infection in vivo. Results showed that 105 and 106 cells mL−1V. harveyi and 106 cells mL−1S. aureus significantly affected plasma hemocyanin concentration, hepatopancreas hemocyanin mRNA and subunits expressions, plasma phenol oxidase (PO), hemocyanin-derived PO (Hd-PO), antibacterial, and bacteriolytic activities during the experiment under bacterial stress, while these parameters did not change remarkably in control group. The concentration of hemocyanin in plasma fluctuated, with a minimum at 12 h and a maximum at 24 h. Moreover, the expression of hemocyanin mRNA peaked at 12 h, while the level of hemocyanin p75 and p77 subunits reached maximum at 24 h. Besides, plasma PO and Hd-PO activities peaked at 24 h, and antimicrobial and bacteriolytic activities peaked at 12 h and 24 h, respectively. In addition, 105 cells mL−1S. aureus had no significant effect on the synthesis of hemocyanin and prophenoloxidase activating (pro-PO) system, but significantly increased antimicrobial activity at 12 h and bacteriolytic activity at 24 h. Therefore, these results suggest that the hemocyanin synthesis was initiated after invasion of pathogen, and the newly synthesized hemocyanin, acted as an immune molecule, can exerts PO activity to regulate the immune defense in L. vannamei in vivo.