The shear-sensitive microalga Heterosigma akashiwo is known to produce brevetoxin-like compounds that are active in voltage-dependent sodium channels. In this work, we present a study on the production of anaesthetic extracts from H. akashiwo biomass obtained in low-shear bioreactors at different growth phases. The photobioreactors (PBRs) used had specific configurations and were operated in such a way as to avoid cellular damage by hydrodynamic stress. Cultures were developed in small static-control flasks and PBRs with volumes ranging from 1.5 L to 200 L. The bioactivity of the produced extracts was assessed in vitro (Neuro-2a cell-based assay) and in vivo (Zebra fish model). Bioactivity depended slightly on the growth phase and culture system, with greater toxicity with the Neuro-2a model when stationary-phase extracts were used. Interestingly, extracts were not cytotoxic against other human cell lines. Steady production of anaesthetic bioactives was observed. In vivo anaesthetic efficacy, assessed with the Zebra fish model, was similar to that of commercial products, and without any observed mortality at 24-h post exposure.