Probiotic Treatment Enhances Pre‐feeding Larval Development and Early Survival in Zebrafish Danio rerio

The growth and development of healthy culture subjects are essential in increasing productivity in the aquaculture industry. A primary determinant of aquatic animal productivity is the ambient microbial population. If an aquatic animal's microbiome is diverse, with bacteria favoring beneficial over pathogenic species, the health and growth of the animal (i.e., fish or crustacean) can be substantially improved. Embryonic and newly hatched Zebrafish Danio rerio larvae were reared in the presence of (1) water from the broodstock culture tank as a control, (2) a probiotic solution containing 19 strains of live lactic acid bacteria (LAB), or (3) an antibiotic (AB) solution with amoxycillin. Developmental parameters were monitored until 10 d postfertilization. Bacteria present in the water and larvae were cultured and identified by sequencing the V4 hypervariable region of bacterial 16S ribosomal RNA. Probiotic-treated larvae showed significant increases in every measured morphological parameter and in survival compared to the controls and AB-treated larvae, including TL, eye development, and swim bladder development before first feeding. Staining with DASPEI (2-(4-[dimethylamino]styryl)-N-ethylpyridinium iodide) produced fluorescence, revealing increased mitochondrial activity in the gastrointestinal tracts of probiotic-treated larvae and reflecting advancement of initial metabolic function. Probiotic-treated larvae showed accelerated yolk absorption, resulting in increased nutrient mobilization and growth. Microbial analyses revealed a greater concentration of bacteria in larvae in response to the probiotic treatment compared to the other two treatments. Species identified in all three treatments included Pseudomonas spp. and Aeromonas spp. (Proteobacteria). The second most diverse and abundant microbiome was seen in controls, whereas AB-treated larvae had the least diverse microbiome. All treatments revealed the presence of proteobacteria, but an AB-resistant pathogenic bacterium (Stenotrophomonas maltophilia) was identified in the AB group. These results reveal that the presence of LAB and other bacteria favorably influenced early larval growth, development, digestive function, and survival in Zebrafish even before the onset of feeding.