Studies on nitric oxide synthase activity in haemocytes of shrimps Fenneropenaeus chinensis and Marsupenaeus japonicus after white spot syndrome virus infection

Abstract

The nitric oxide synthase (NOS) activity in the haemocytes of shrimps Fenneropenaeus chinensis (Osbeck) and Marsupenaeus japonicus (Bate) was studied after white spot syndrome virus (WSSV) infection to determine its characteristics in response to virus infection. First, the NOS activity in haemocytes of shrimps was determined by the means of NBT reduction and changes in cell conformation. And the variations of NOS activity in shrimps after challenge with WSSV intramuscularly were evaluated through the analysis of l-citrulline and total nitrite/nitrate (both as NO derivates) concentrations. The result showed that NOS activity in the haemocytes of F. chinensis increased slightly from 0 to 12 h postchallenge, indicated by the variations of l-citrulline (from 11.15 ± 0.10 to 12.08 ± 0.64 μM) and total nitrite/nitrate concentrations (from 10.45 ± 0.65 to 12.67 ± 0.52 μM). Then it decreased sharply till the end of the experiment (84 h postchallenge), the concentrations of l-citrulline and total nitrite/nitrate at 84 h were 1.58 ± 0.24 and 2.69 ± 0.70 μM, respectively. The LPS-stimulated NOS activity kept constant during the experiment. However, in M. japonicus, the NOS activity kept increasing during the first 72 h postchallenge, the concentrations of l-citrulline and total nitrite/nitrate increased from 7.82 ± 0.77 at 0 h to 10.79 ± 0.50 μM at 72 h, and from 8.98 ± 0.43 at 0 h to 11.20 ± 0.37 μM at 72 h, respectively. Then it decreased till the end of the experiment (216 h postchallenge), and the concentrations of l-citrulline and total nitrite/nitrate at 216 h were 5.66 ± 0.27 and 4.68 ± 0.16 μM, respectively. More importantly, an apparent increase of LPS-stimulated NOS activity was observed in M. japonicus at 48 h postchallenge, which was about 4 times higher than that in the control group of health shrimps. In correspondence with the difference of NOS activity between the two species of shrimps, the cumulative mortalities of the shrimps were also different. All shrimps of F. chinensis in the mortality experiment died in 66 h, much more quickly than M. japonicus, whose accumulative mortality reached 100% after 240 h. Data here reported let us hypothesize that NOS activity in the haemocytes of shrimps F. chinensis and M. japonicus responses to WSSV infection differently, and this might be one of the reasons for the different susceptibility of F. chinensis and M. japonicus to WSSV infection.