Production of extracts with anaesthetic activity from the culture of Heterosigma akashiwo in pilot-scale photobioreactors

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

Sublethal effect of the toxic dinoflagellate Karlodinium veneficum on early life stages of zebrafish (Danio rerio)

Dinoflagellates of the genus Karlodinium are ichthyotoxic species that produce toxins including karlotoxins and karmitoxins. Karlotoxins show hemolytic and cytotoxic activities and have been associated with fish mortality. This study evaluated the effect of toxins released into the environment of Karlodinium veneficum strain K10 (Ebro Delta, NW Mediterranean) on the early stages of Danio rerio (zebrafish). Extracts of the supernatant of K10 contained the mono-sulfated KmTx-10, KmTx-11, KmTx-12, KmTx-13, and a di-sulfated form of KmTx-10. Total egg mortality was observed for karlotoxin concentration higher than 2.69 μg L−1. For 1.35 μg L−1, 87% of development anomalies were evidenced (all concentrations were expressed as KmTx-2 equivalent). Larvae of 8 days postfertilization exposed to 1.35 µg L−1 presented epithelial damage with 80% of cells in the early apoptotic stage. Our results indicate that supernatants with low concentration of KmTxs produce both lethal and sublethal effects in early fish stages. Moreover, apoptosis was induced at concentrations as low as 0.01 μg L−1. This is of great relevance since detrimental long-term effects due to exposure to low concentrations of these substances could affect wild and cultured fish

Physiological Response of Atlantic Salmon (Salmo salar) to Long-Term Exposure to an Anesthetic Obtained from Heterosigma akashiwo

Despite the invaluable role of anesthetics as a tool for ensuring animal welfare in stressful situations, there is currently a lack of anesthetic drugs that meet the requirements of intensive aquaculture. In response to the growing interest in anesthetic substances of natural origin, this study evaluated the physiological and health impact of an anesthetic based on an extract of the microalga Heterosigma akashiwo on juvenile salmon (Salmo salar) exposed for a period of 72 h. To simulate a condition closer to reality where fish are subjected to stimuli (e.g., transport), the animals were exposed to 50 mg L−1 of algal extract and to physical stress. Functional, physiological, and histological parameters were evaluated in blood and tissues at different sampling periods (0, 24, and 72 h). There was no mortality and the induction and recovery times observed were within the established criteria for anesthetic efficacy. The anesthetic extract did not induce any side effects, such as stress or metabolic damage, indicating that this extract is a viable option for supporting fish welfare during deleterious events. This study provides information to support that the anesthetic extract tested, derived from H. akashiwo, is a promising candidate drug for operations requiring sedation (e.g., Salmonid transport)