Managing input C/N ratio to reduce the risk of Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks in biofloc systems – A laboratory study

Abstract

Biofloc systems are microbial mature environments that are potentially less conducive disease outbreaks. We hypothesized that the way in which biofloc microbial communities are managed determines the level of disease protection. To investigate such hypothesis, Litopenaeus vannamei post-larvae were cultured for 21 days in biofloc environments created by different water management procedures. Five different types of bioflocs were created: autotrophic bioflocs without probiotics, autotrophic bioflocs with probiotics, heterotrophic bioflocs without probiotics, heterotrophic bioflocs with probiotics, and a flow-through system as a control. Heterotrophic bioflocs were obtained by daily addition of carbon (glucose) at an estimated C/N ratio of 18 throughout the experiment. For autotrophic bioflocs this input of carbon was applied only to start up the system and upon appearance of bioflocs (TSS > 100 mg L−1) and a drop in total ammonium nitrogen concentration below 0.05 mg L−1, carbon dosing was stopped. Bioflocs cultured with addition of probiotics received a 0.5 ppm dose every 48 hours. After 21-d culture period, a 96 h challenge test was performed with a Vibrio parahaemolyticus strain known to cause AHPND. For each biofloc type, this challenge was performed in three different approaches: 1- Shrimp were taken out of their biofloc tanks and challenged by applying new seawater; 2-Shrimp from biofloc tanks were challenged in their respective biofloc suspensions; and 3- Non-experimental shrimp, randomly selected from a recirculation (RAS) system were challenged in the types of biofloc suspensions. Mortality was high when shrimp were challenged in new seawater, independent of treatment. When challenged in their respective biofloc suspensions shrimp survival was the highest in heterotrophic bioflocs with and without probiotic supplementation and the autotrophic bioflocs with probiotics, whereas shrimp survival in autotrophic bioflocs without probiotics was 50%. These results were similar when non-experimental shrimp originating from a RAS system were challenged in these biofloc suspensions. Taken together, results suggest that bioflocs as such can decrease the impact of a Vibrio parahaemolyticus challenge and that this protection depends on the operational parameters of the biofloc system. Moreover, probiotics can be used to complement the protective effect of bioflocs. This information reinforces the importance of microbial community management as a tool to reduce the risk of disease and establish highly biosecure systems.