Exposure to probiotics and β-1,3/1,6-glucans in larviculture modifies the immune response of Penaeus vannamei juveniles and both the survival to White Spot Syndrome Virus challenge and pond culture

Abstract

The effect of the inclusion of probiotics (Vibrio alginolyticus) and β-1,3/1,6-glucans in Penaeus vannamei larviculture was evaluated by measuring the immune response and survival of shrimp juveniles subjected to white spot syndrome virus (WSSV) challenge tests (per os) and pond culture. Treatments were designed to contrast the probiotic factor (inclusion vs non-inclusion) and β-1,3/1,6-glucans supplementation in various larvae stages; starting from early stage (Zoea II), to middle stage (PL 12), late stage (15 days pre-challenge) or non-supplemented. In larviculture, the highest survival was obtained in animals treated with β-1,3/1,6-glucans from Zoea II. The use of probiotics enhanced survival during the first 0–52 h post-WSSV challenge period. During 56–156 h post-WSSV challenge period, interactions were observed between β-1,3/1,6-glucans×time and β-1,3/1,6-glucans×time×probiotics. Significant differences in final survival rates between treatments were not observed. In the second WSSV challenge, immune parameters were analysed. Significant interaction between probiotics and β-1,3/1,6-glucans was observed for plasmatic protein (PP) concentration, super oxide anion (O2−) generation, antibacterial activity (AA), and total haemocyte count (THC). The use of probiotics in larviculture had a negative effect on the PP, but increased the AA and THC, while β-1,3/1,6-glucans had a negative effect on the O2− generation. The most relevant results were obtained from treatments T2 (probiotics in larviculture, β-1,3/1,6-glucans from Z II) and T4 (probiotics in larviculture, β-1,3/1,6-glucans 15 days before challenge). Treatment T2 presented the highest survival rate in larviculture. After WSSV infection, the animals of this treatment displayed resistance to the virus, a strong AA and increase of THC. Treatment T4 increased the amount of PP, increased the O2− generation and THC. Histological analysis showed that the animals of treatment T2 and T4 were able to limit the spread of the virus during the first hour after challenge with WSSV. The survivors from treatments T2 and T4 had a high THC, accompanied by a lack of white spot disease (WSD) injuries. A bioassay was carried out under farm conditions during the warm-rainy season using larvae from treatments T2 and T4. The animals were stocked at 18 animals/m2 in earth ponds of 0.20 ha (three ponds/larvae kind). WSD outbreak was not detected, and the survival was significantly higher in ponds stocked with larvae from treatment T4 (70±3%) than in ponds stocked with larvae from treatment T2 (49±9%).