Multi-strain probiotics enhance immune responsiveness and alters metabolic profiles in the New Zealand black-footed abalone (Haliotis iris)

We assessed whether dietary administration of a multi-strain probiotic (Exiguobacterium JHEb1, Vibrio JH1 and Enterococcus JHLDc) lead to enhanced immune responsiveness in juvenile New Zealand black-footed abalone (Haliotis iris). Two groups of abalone were fed (1% body weight per day) over a four-month period with different diets. The control diet consisted of a standard commercial pellet feed (AbMax 16), whereas the treatment diet was additionally enriched with the probiotic mix. At the end of the experiment, probiotic-fed animals showed improved growth compared with control-fed abalone in length (32.3% vs 22.3%), width (31.9% vs 20.7%) and wet weight (109.6% vs 72.8%), respectively. Haemolymph sampling was conducted at the beginning of the experiment and after 2 and 4 months. Haemolymph samples were analysed for total haemocyte count (THC) and viability, presence of apoptotic cells and production of Reactive Oxygen Species (ROS). Compared with control abalone, probiotic-fed abalone had significantly higher THC (1.9 x 10(6) vs 5.6 x 10(6) cells), higher viability (90.8% vs 75.6%), higher percentage of ROS-positive cells (19.4% vs 0.5%) and higher numbers of non-apoptotic cells (88.0% vs 78.0%), respectively. These results indicate that the probiotic-enriched diet enhanced the immunostimulatory mechanisms, with a simultaneous low-level up-regulation of ROS production as a priming mechanism of the antibacterial defence system. Metabolomics-based profiling of foot muscle tissue additionally revealed that probiotic-fed abalone differentially expressed 17 unique metabolites, including amino acids, fatty acids and TCA cycle related compounds. These data suggest that the probiotic-supplemented diet can also alter central carbon metabolic processes, which may improve the survival, as well as the growth of abalone.