Table 1.
Accession numbers and primer sequences for RT-qPCR.
Figure 1.
Quantification of mRNA encoding shrimp immune-related proteins following NLHS.
Post-larvae were exposed to 30 min heat shock from 28°C to 30°C, 32°C, 34°C, 36°C and 38°C, then transferred to 28°C for 8 h. mRNAs encoding immune related proteins were quantified by RT-qPCR with beta-actin as reference. The error bars represent SD from 3 replicates. proPO, prophenoloxidase; He, hemocyanin; Pe, peroxinectin; Crus, crustin; Pen, penaedin; HS, heat shock. Asterisks denote statistically significant differences between values obtained for control and heat shocked post-larvae (P<0.05). The figure is a representation from two separate experiments.
Figure 2.
NLHS increased shrimp Hsp70 mRNA.
Post-larvae were exposed to 30 min heat shock from 28°C to 30°C, 32°C, 34°C, 36°C and 38°C, then transferred to 28°C for 8 h. Hsp70 mRNA was quantified by RT-qPCR, with beta-actin as reference. Bars represent the fold difference of Hsp70 mRNA with comparison to the non-heated control. The error bars represent the SD from 3 replicates. Asterisks denote statistically significant differences between values obtained for control and heat shocked post-larvae (P<0.05). The figure is a representation from two separate experiments.
Figure 3.
NLHS increased Hsp70 in L. vannamei post-larvae.
Protein extracts were resolved by electrophoresis in SDS polyacrylamide gels and either stained with Coomassie Biosafe (A) or blotted to polyvinylidene fluoride membranes and incubated with antibody to Hsp70 (B). Approximately 50 µg of protein was loaded in each lane. 28, non-heat shocked post-larvae heat shock at 30°C, 32°C, 34°C, 36°C and 38°C was for 30 min with recovery at 28°C for 8 h H-Hsp70: Human Hsp70 recombinant protein M: molecular mass standards in kDa. The figure is a representation from two separate experiments.
Figure 4.
NLHS enhanced Hsp70 production in L. vannamei.
Post-larvae were exposed to 30 min heat shock from 28°C to 30°C, 32°C, 34°C, 36°C and 38°C, then transferred to 28°C for 8 h. Quantification of Hsp70 was by ELISA. A standard curve, constructed with Human Hsp70 recombinant protein was used to convert ELISA readings obtained with shrimp tissue protein extracts to Hsp70 content. Bars represent the fold difference of Hsp70 quantity in comparison to a non-heated control. The error bars represent the SD from 3 replicates. Asterisks denote statistically significant differences between values obtained for control and heat shocked post-larvae (P<0.05). The figure is a representation from two separate experiments.
Table 2.
Survival (%) of L. vannamei post-larvae after 24, 48, 72 and 96 h challenges with 1×101, 1×103, 1×105 and 1×107 V. harveyi/mL.
Table 3.
Average survival (%) of heat shocked L. vannamei post-larvae after challenge for 24, 48 and 72 h with 1×107 V. harveyi/ml.