Effect of high water temperature (33 °C) on the clinical and virological outcome of experimental infections with white spot syndrome virus (WSSV) in specific pathogen-free (SPF) Litopenaeus vannamei

Abstract

White spot syndrome virus (WSSV) is the most lethal pathogen of cultured shrimp. Previous studies done with undefined WSSV titers showed that high water temperature (32–33 °C) reduced/delayed mortality of WSSV-infected shrimp. This study evaluated the effect of high water temperature on the clinical and virological outcome of a WSSV infection under standardized conditions. Groups of specific pathogen-free Litopenaeus vannamei were challenged either by intramuscular or oral routes with a low (30 SID50) or a high (10,000 SID50) virus titer. Shrimp were kept (i) continuously at 27 °C, (ii) 30 °C or (iii) 33 °C; (iv) maintained at 33 °C before challenge and 27 °C afterwards, or (v) kept at 27 °C before challenge and 33 °C afterwards. Shrimp were maintained at the respective temperatures for 120 h before challenge and 120–144 h post challenge (hpc). Gross signs and mortality were monitored every 12 h until the end of the experiment. Dead and surviving shrimp were screened for WSSV infection (VP28-positive cells) by indirect immunofluorescence (IIF). Shrimp kept continuously at 27 °C or 30 °C, or switched to 27 °C post challenge developed gross signs within 24 hpc, first mortalities at 36–60 hpc and 100% cumulative mortality between 60 and 144 hpc depending on the virus titer. All dead shrimp were WSSV-positive. In contrast, shrimp kept at 33 °C continuously or after WSSV challenge showed no signs of disease and low mortalities (0–30%) regardless of the virus titer. Dead and surviving shrimp were WSSV-negative. Further, early virus replication was studied in two groups of shrimp: one maintained at 27 °C before and after challenge and one switched from 27 °C to 33 °C after challenge with 10,000 SID50. Immunohistochemistry (IHC) analysis showed that WSSV-positive cells were first displayed at 12 hpc in shrimp kept at 27 °C and by 24 hpc the infection became systemic. In contrast, shrimp kept at 33 °C did not display WSSV-positive cells at 12 or 24 hpc. This work confirms previous reports that high water temperature prevents the onset of disease and significantly reduces mortality of WSSV-inoculated shrimp regardless of the route of inoculation or virus titer used. This strategy may have practical applications to control WSSV in tropical shrimp farming countries.